Native Australian Marsupials Research Articles

The missing toxic link: Exposure of non-target native marsupials to second-generation anticoagulant rodenticides (SGARs) suggest a potential route of transfer into apex predators

Anticoagulant rodenticides (ARs) are used globally to control rodent pests. Second-generation anticoagulant rodenticides (SGARs) persist in the liver and pose a significant risk of bioaccumulation and secondary poisoning in predators, including species that do not generally consume rodents. As such, there is a clear need to understand the consumption of ARs, particularly SGARs, by non-target consumers to determine the movement of these anticoagulants through ecosystems. We collected and analysed the livers from deceased common brushtail possums (Trichosurus vulpecula) and common ringtail possums (Pseudocheirus peregrinus), native Australian marsupials that constitute the main diet of the powerful owl (Ninox strenua), an Australian apex predator significantly exposed to SGAR poisoning. ARs were detected in 91 % of brushtail possums and 40 % of ringtail possums. Most of the detections were attributed to SGARs, while first-generation anticoagulant rodenticides (FGARs) were rarely detected. SGAR concentrations were likely lethal or toxic in 42 % of brushtail possums and 4 % of ringtail possums with no effect of age, sex, or weight detected in either species. There was also no effect of the landscape type possums were from, suggesting SGAR exposure is ubiquitous across landscapes. The rate of exposure detected in these possums provides insight into the pathway through which ARs are transferred to one of their key predators, the powerful owl. With SGARs entering food-webs through non-target species, the potential for bioaccumulation and broader secondary poisoning of predators is significantly greater and highlights an urgent need for routine rodenticide testing in non-target consumers that present as ill or found deceased. To limit their impact on ecosystem stability the use of SGARs should be significantly regulated by governing agencies.

Outfoxing the fox: Effect of prey odor on fox behavior in a pastoral landscape

AbstractInvasive mammalian predators have had a devastating effect on native species globally. The European red fox (Vulpes vulpes) is one such species where it has been introduced in Australia. A novel but unexplored tactic to reduce the impact of mammalian predators is the use of unrewarded prey odors to undermine the effectiveness of olfactory hunting behavior. To test the viability of unrewarded prey odors in an applied setting we investigated how foxes responded to the odors of three different prey species. We used the odors of two locally extinct native Australian marsupials; the eastern quoll (a smaller carnivore) and eastern bettong (a fungivore), and the European rabbit, an introduced herbivore. Conducting our research over a period of 3 weeks in a pastoral environment in South‐eastern Australia, we used video observations of foxes' behaviors, as they encountered the different odors. We found a reduction in the number of fox visits to bettong odors in the third week. In contrast, we observed a sustained number of visits to rabbit odors. Foxes also spent more time investigating rabbit odors and displayed longer durations of vigilance behavior at quoll odors. Our results support the hypothesis that the exposure of wild foxes to unrewarded odors of novel prey species can reduce their interest in these odors, which might translate to a reduction in predation pressure. Our results also suggest, however, that olfactory pre‐exposure may not be as effective at reducing fox interest in a competitor species' odor.

Novel hepaci- and pegi-like viruses in native Australian wildlife and non-human primates.

The Flaviviridae family of positive-sense RNA viruses contains important pathogens of humans and other animals, including Zika virus, dengue virus, and hepatitis C virus. The Flaviviridae are currently divided into four genera—Hepacivirus, Pegivirus, Pestivirus, and Flavivirus—each with a diverse host range. Members of the genus Hepacivirus are associated with an array of animal species, including humans, non-human primates, other mammalian species, as well as birds and fish, while the closely related pegiviruses have been identified in a variety of mammalian taxa, also including humans. Using a combination of total RNA and whole-genome sequencing we identified four novel hepaci-like viruses and one novel variant of a known hepacivirus in five species of Australian wildlife. The hosts infected comprised native Australian marsupials and birds, as well as a native gecko (Gehyra lauta). From these data we identified a distinct marsupial clade of hepaci-like viruses that also included an engorged Ixodes holocyclus tick collected while feeding on Australian long-nosed bandicoots (Perameles nasuta). Distinct lineages of hepaci-like viruses associated with geckos and birds were also identified. By mining the SRA database we similarly identified three new hepaci-like viruses from avian and primate hosts, as well as two novel pegi-like viruses associated with primates. The phylogenetic history of the hepaci- and pegi-like viruses as a whole, combined with co-phylogenetic analysis, provided support for virus-host co-divergence over the course of vertebrate evolution, although with frequent cross-species virus transmission. Overall, our work highlights the diversity of the Hepacivirus and Pegivirus genera as well as the uncertain phylogenetic distinction between.

Identification of A Novel Picorna-Like Virus, Burpengary Virus, that is Negatively Associated with Chlamydial Disease in the Koala.

Koalas (Phascolarctos cinereus) are native Australian marsupials whose populations are in decline from a range of threats. Infectious diseases caused by the bacterium Chlamydia pecorum and other pathogens are of particular concern. We analysed 26 poly-A selected RNA-sequencing libraries from a data set designed to study the immune response of koalas to ocular chlamydial infection. Using virus discovery techniques, we identified the coding-complete genome sequence of a novel picorna-like virus, denoted Burpengary virus, that was most common in south-east Queensland. Notably, abundance measurements of the virus across all 26 libraries revealed an inverse relationship between abundance and ocular disease in koalas, suggesting that the co-infection of Burpengary virus and Chlamydia pecorum is inhibited.

Mitochondrial genome sequencing reveals potential origins of the scabies mite Sarcoptes scabiei infesting two iconic Australian marsupials

BackgroundDebilitating skin infestations caused by the mite, Sarcoptes scabiei, have a profound impact on human and animal health globally. In Australia, this impact is evident across different segments of Australian society, with a growing recognition that it can contribute to rapid declines of native Australian marsupials. Cross-host transmission has been suggested to play a significant role in the epidemiology and origin of mite infestations in different species but a chronic lack of genetic resources has made further inferences difficult. To investigate the origins and molecular epidemiology of S. scabiei in Australian wildlife, we sequenced the mitochondrial genomes of S. scabiei from diseased wombats (Vombatus ursinus) and koalas (Phascolarctos cinereus) spanning New South Wales, Victoria and Tasmania, and compared them with the recently sequenced mitochondrial genome sequences of S. scabiei from humans.ResultsWe found unique S. scabiei haplotypes among individual wombat and koala hosts with high sequence similarity (99.1% - 100%). Phylogenetic analysis of near full-length mitochondrial genomes revealed three clades of S. scabiei (one human and two marsupial), with no apparent geographic or host species pattern, suggestive of multiple introductions. The availability of additional mitochondrial gene sequences also enabled a re-evaluation of a range of putative molecular markers of S. scabiei, revealing that cox1 is the most informative gene for molecular epidemiological investigations. Utilising this gene target, we provide additional evidence to support cross-host transmission between different animal hosts.ConclusionsOur results suggest a history of parasite invasion through colonisation of Australia from hosts across the globe and the potential for cross-host transmission being a common feature of the epidemiology of this neglected pathogen. If this is the case, comparable patterns may exist elsewhere in the ‘New World’. This work provides a basis for expanded molecular studies into mange epidemiology in humans and animals in Australia and other geographic regions.

Molecular evidence of Chlamydia pecorum and arthropod-associated Chlamydiae in an expanded range of marsupials

The order Chlamydiales are biphasic intracellular bacterial pathogens infecting humans and domesticated animals. Wildlife infections have also been reported, with the most studied example being Chlamydia pecorum infections in the koala, an iconic Australian marsupial. In koalas, molecular evidence suggests that spill-over from C. pecorum infected livestock imported into Australia may have had a historical or contemporary role. Despite preliminary evidence that other native Australian marsupials also carry C. pecorum, their potential as reservoirs of this pathogen and other Chlamydia-related bacteria (CRBs) has been understudied. Mucosal epithelial samples collected from over 200 native Australian marsupials of different species and geographic regions across Australia were PCR screened for Chlamydiales. Previously described and genetically distinct C. pecorum genotypes and a range of 16S rRNA genotypes sharing similarity to different CRBs in the broader Chlamydiales order were present. One 16S rRNA Chlamydiales genotype recently described in Australian ticks that parasitise native Australian marsupials was also identified. This study provides further evidence that chlamydial infections are widespread in native fauna and that detailed investigations are required to understand the influence these infections have on host species conservation, but also whether infection spill-over plays a role in their epidemiology.

To bask or not to bask: Behavioural thermoregulation in two species of dasyurid, Phascogale calura and Antechinomys laniger

Torpor is an important physiological process used to reduce body temperature and thus energy needs in mammals and birds. Rewarming from torpor can be costly. In some small mammals basking is used to reduce the cost of rewarming from torpor. The use and duration of basking was measured in two native Australian marsupials, the red-tailed phascogale (Phascogale calura) and kultarr (Antechinomys laniger), using a heat rock. Animals were observed via video cameras to determine whether they used basking behaviours and whether this behaviour was affected by food availability. Both species became accustomed to the heat rock as time progressed. Absence of food increased basking duration and frequency in some individuals but not all animals. Generally kultarrs basked for longer periods than phascogales, and kultarrs would bask at all hours of the day while phascogales showed a strict nocturnal regime (1900–0700h). The results demonstrate that basking may be an important behavioural adaptation of the kultarr; while the phascogale requires further research.

Detection ofCoxiella burnetiiDNA in Wildlife and Ticks in Northern Queensland, Australia

Wild animals and the tick species that feed on them form the natural transmission cycle and reservoir of Coxiella burnetii. The objective of this study was to determine whether C. burnetii was present in the blood of host animals and their ticks in northern Queensland, Australia. Three genomic targets were detected using real-time PCR assays-the Coxiella-specific outer membrane protein coding gene (Com1), the multicopy insertion element (IS1111), and the isocitrate dehydrogenase gene (Icd). Quantification of the single-copy targets identified a range of 1.48×10(1) to 4.10×10(3) C. burnetii genome equivalents per microliter in the ticks tested. The detection of Coxiella based on the presence of the genomic targets indicated the occurrence of C. burnetii in both the ticks and whole blood of a variety of native Australian marsupials and confirms these animals are capable of acting as reservoirs of Q fever in northern Queensland.

Carriage of Shiga-toxigenic Escherichia coli by native marsupials in Australia

Shiga-toxigenic strains of Escherichia coli (STEC) are zoonotic pathogens with human health, meat processing and trade impacts. Cattle are the principal reservoirs of STEC, although other animals can be carriers. The STEC status of Australian native marsupials has not been formatively described to date. The aim of the current study was to investigate carriage of STEC by native Australian marsupials in Southeast Queensland. Faeces from a variety of marsupial species, stratified by gastrointestinal morphology and dietary type, were screened for stx1, stx2 and other STEC virulence genes by PCR. Positive samples were cultured to isolate STEC for characterisation. A number of macropods from both captive and wild habitats had evidence of STEC in their faeces. Rates of stx carriage by macropods (8.6%) were comparable, though generally low, compared to cattle. Eastern grey kangaroos had the highest rate of stx presence in faeces (10.3%). Hindgut-fermenting and monogastric marsupials had no evidence of STEC shedding. Based on virulence marker possession and serotype, the human pathogenic potential of isolates was low. This is the first report of Australian marsupials carrying STEC. Australian native macropods may act as reservoirs for STEC strains, but the potential significance to public health and/or livestock epidemiology remains questionable.

Identification of novel trypanosome genotypes in native Australian marsupials

In the present study, the occurrence and molecular phylogeny of trypanosome parasites were studied in both wild and captive marsupials from Western Australia and Queensland. Blood samples were screened by PCR at the 18S rDNA locus, and the glycosomal glyceraldehyde phosphate dehydrogenase gene.Overall, 5.3% of the blood samples were positive at the 18S rDNA locus. All positives belonged to wild-captured Western Australian individuals, where trypanosome-specific DNA was detected in 9.8% of the screened samples from wild marsupials, in common brushtail possums, and woylies. The detection rate of trypanosome DNA in these two host species was 12.5% and 20%, respectively.Phylogenetic analyses based on two loci, indicated that the possum-derived trypanosome isolates were genetically distinct, and most closely related to the Australian marsupial trypanosomes H25 from a kangaroo, and BRA2 from a bush rat. This is the first study to genetically characterise trypanosome isolates from possums.The analysis of the woylie-derived isolates demonstrated that this marsupial host can harbour multiple genotypes within the same geographical location and furthermore multiple genotypes within the same host, indicative of mixed infections. All the woylie-derived genotypes grouped with trypanosomes found in Australian marsupials, suggesting that they are more likely to belong to an endemic or Australasian trypanosome species.This is the first study to genetically characterise trypanosome isolates from possums (Trichosurus vulpecula). Although the clinical significance of these infections is currently unknown, the identification of these novel sequences may support future investigations on transmission, threats to endangered wildlife, and evolutionary history of the genus Trypanosoma.

Predator odour does not influence trappability of southern brown bandicoots (Isoodon obesulus) and common brushtail possums (Trichosurus vulpecula)

Predators cause changes in the behaviour of many prey species. This study investigated whether trappability of wild southern brown bandicoots (Isoodon obesulus) and common brushtail possums (Trichosurus vulpecula) was influenced by odour cues suggesting the presence of potential predators. Trapping success was compared between traps with predator scents and controls in two different experiments. The first measured trapping success of single clean traps, traps with unfamiliar herbivore (horse) odour and traps scented with predator (fox, cat, quoll and dingo) odours, while the second offered three choices (fox, dingo and clean traps) simultaneously. Frequency of capture of bandicoots and possums was not influenced by odour, mass or sex in either experiment. The lack of avoidance response observed in this study suggests that neither southern brown bandicoots nor common brushtail possums alter foraging behaviour in response to olfactory cues that suggest the presence of exotic or Australian predators. This is consistent with previous studies, which indicate that native Australian marsupials do not avoid predator odours.