Streamlining Disease Risk Analysis for Wildlife Using the Shark Bay Bandicoot as a Model.

Disease risk analysis (DRA) is a process for identifying significant disease risks and proposing measures to mitigate those risks. Although numerous methodologies for DRA exist, the IUCN Disease Risk Analysis Manual Jakob-Hoff et al. (World Organisation for Animal Health, Paris, pp 160, 2014) remains the gold standard for wild animal translocations. In some cases, however, constraints of time or resources demand compromises on the ideal methodology, and a cost-benefit assessment is required to determine the best approach. We propose a methodology modified from Jakob-Hoff et al. (World Organisation for Animal Health, Paris, pp 160, 2014) and Sainsbury and Vaughan-Higgins (Conserv Biol 26:442-452, 2012), using translocations of the Shark Bay bandicoot (SBB) (Perameles bougainville) as an example. In this study, 44 hazards were identified and described for Peramelidae species. We used hazard prioritization and "scoping" to develop a shortlist of hazards for detailed risk assessment, which excluded 35 of these hazards from further assessment. This approach enabled timely, efficient and cost-effective completion of the DRA while maintaining transparent evaluation of all disease risks. We developed a disease risk management strategy for SBB based on structured, evidence-based analysis of current information and established biosecurity practices and disease screening recommendations for future translocations. Our approach demonstrates a practical process for DRA and risk mitigation, which delivered management outcomes despite limited resources, variable knowledge of disease epidemiology and uncertain translocation pathways for the target species. Limitations are acknowledged, and further research will aim to objectively test this methodology compared to other available methods.

The Importance of Wildlife Disease Monitoring as Part of Global Surveillance for Zoonotic Diseases: The Role of Australia.

Australia has a comprehensive system of capabilities and functions to prepare, detect and respond to health security threats. Strong cooperative links and coordination mechanisms exist between the human (public health) and animal arms of the health system in Australia. Wildlife is included in this system. Recent reviews of both the animal and human health sectors have highlighted Australia's relative strengths in the detection and management of emerging zoonotic diseases. However, the risks to Australia posed by diseases with wildlife as part of their epidemiology will almost certainly become greater with changing land use and climate change and as societal attitudes bring wildlife, livestock and people into closer contact. These risks are not isolated to Australia but are global. A greater emphasis on wildlife disease surveillance to assist in the detection of emerging infectious diseases and integration of wildlife health into One Health policy will be critical in better preparing Australia and other countries in their efforts to recognize and manage the adverse impacts of zoonotic diseases on human health. Animal and human health practitioners are encouraged to consider wildlife in their day to day activities and to learn more about Australia's system and how they can become more involved by visiting www.wildlifeheathaustralia.com.au.

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.

Prevalence, genetic diversity and potential clinical impact of blood-borne and enteric protozoan parasites in native mammals from northern Australia.

A molecular survey was conducted to provide baseline information on the prevalence, genetic diversity and potential clinical impacts of blood-borne and enteric protozoans in native wild mammals from the Northern Territory (NT). A total of 209 blood and 167 faecal samples were collected from four target species; the northern brown bandicoot (Isoodon macrourus), common brushtail possum (Trichosurus vulpecula), northern quoll (Dasyurus hallucatus) and brush-tailed rabbit-rat (Conilurus penicillatus). Blood samples were screened by PCR at the 18S rRNA gene for trypanosomes, piroplasms and haemogregarines, with faecal samples tested for Cryptosporidium spp. at the 18S rRNA locus, and for Giardia spp. at the glutamate dehydrogenase (gdh) and 18S rRNA loci. The potential clinical impact was investigated by associating clinical, haematological and biochemical parameters with presence or absence of infection. Overall, 22.5% (95% CI: 17.0-28.8%) of the animals tested were positive for haemoprotozoans. Trypanosomes were found in 26.6% (95% CI: 18.7-35.7%) of the bandicoots and were identified as Trypanosoma vegrandis G6, except for one unique genotype, most similar to T. vegrandis G3 (genetic distance=7%). The prevalence of trypanosomes in possums was 23.7% (95% CI: 11.4-40.2%), and the genotypes identified clustered within the T. noyesi clade. The presence of Babesia sp. and Hepatozoon sp. was confirmed in bandicoots only, both at a prevalence of 9.7% (95% CI: 2.7-9.2%). The total prevalence of intestinal protozoan parasites observed was relatively low (3%; 95% CI: 1.0-6.9%). No evidence of clinical disease associated with protozoan parasitic infection was observed, however bandicoots positive for Trypanosoma exhibited a significantly lower packed cell volume (PCV) compared to negative bandicoots (p=0.046). To the authors' knowledge, this is the first research conducted in the NT to characterise protozoan parasites in threatened native mammals using both molecular and morphological tools; and to assess the potential clinical impacts of these agents. The absence of clear signs of major morbidity in infected animals seems to exclude a direct association between infections with these agents and possible population decline events in northern Australian native mammals. However until the cause(s) of population decline are ascertained for each individual mammal species, further studies are required. The outcome of the present investigation may be used to inform wildlife conservation and zoonotic disease programs.

DEVELOPMENT OF REFERENCE RANGES FOR PLASMA TOTAL CHOLINESTERASE AND BRAIN ACETYLCHOLINESTERASE ACTIVITY IN FREE-RANGING CARNABY'S BLACK-COCKATOOS (CALYPTORHYNCHUS LATIROSTRIS).

Published avian reference ranges for plasma cholinesterase (ChE) and brain acetylcholinesterase (AChE) are numerous. However, a consistently reported recommendation is the need for species- and laboratory-specific reference ranges because of variables, including assay methods, sample storage conditions, season, and bird sex, age, and physiologic status. We developed normal reference ranges for brain AChE and plasma total ChE (tChE) activity for Carnaby's Black-Cockatoos (Calyptorhynchus latirostris) using a standardized protocol (substrate acetylthiocholine at 25 C). We report reference ranges for brain AChE (19-41 µmol/min per g, mean 21±6.38) and plasma tChE (0.41-0.53 µmol/min per mL, mean 0.47±0.11) (n=15). This information will be of use in the ongoing field investigation of a paresis-paralysis syndrome in the endangered Carnaby's Black-Cockatoos, suspected to be associated with exposure to anticholinesterase compounds and add to the paucity of reference ranges for plasma tChE and brain AChE in Australian psittacine birds.

Emerging infectious diseases in free-ranging wildlife-Australian zoo based wildlife hospitals contribute to national surveillance.

Emerging infectious diseases are increasingly originating from wildlife. Many of these diseases have significant impacts on human health, domestic animal health, and biodiversity. Surveillance is the key to early detection of emerging diseases. A zoo based wildlife disease surveillance program developed in Australia incorporates disease information from free-ranging wildlife into the existing national wildlife health information system. This program uses a collaborative approach and provides a strong model for a disease surveillance program for free-ranging wildlife that enhances the national capacity for early detection of emerging diseases.

Health evaluation of free-ranging eastern bettongs (Bettongia gaimardi) during translocation for reintroduction in Australia.

Sixty (19 male, 41 female) free-ranging adult eastern bettongs (Bettongia gaimardi) were captured in Tasmania and translocated to the Australian Capital Territory between July 2011 and September 2012 for reintroduction into fenced, predator-proof reserves. The bettongs were anesthetized for physical examination and screened for selected diseases during translocation. Reference ranges for hematologic and biochemical parameters were determined. Two bettongs had detectable antibodies to the alphaherpesviruses macropodid herpesvirus 1 and macropodid herpesvirus 2 by serum neutralization assay. A novel gammaherpesvirus was detected, via PCR, from pooled swabs collected from the nasal, conjunctival, and urogenital tract mucosa of four other bettongs. Sera from 59 bettongs were negative for antibodies to Toxoplasma gondii as assessed by both the modified agglutination test and the direct agglutination test (n = 53) or by the modified agglutination test only (n = 6). Rectal swabs from 14 bettongs were submitted for bacterial culture and all were negative for Salmonella serovars. Ectoparasites identified on the bettongs included fleas (Pygiopsylla zethi, Stephanocircus harrisoni), a louse (Paraheterodoxous sp.), mites (Guntheria cf. pertinax, Haemolaelaps hatteni, a suspected protonymph of Thadeua sp., Cytostethum tasmaniense, Cytostethum intermedium, Cytostethum thetis, Cytostethum wallabia), and ticks (Ixodes cornuatus, Ixodes trichosuri, Ixodes tasmani). An intraerythrocytic organism morphologically consistent with a Theileria species was identified in blood smears from four bettongs. These data provide baseline health and disease information for free-ranging eastern bettongs that can be used for the conservation management of both the source and translocated populations.

The first complete papillomavirus genome characterized from a marsupial host: a novel isolate from Bettongia penicillata.

The first fully sequenced papillomavirus (PV) of marsupials, tentatively named Bettongia penicillata papillomavirus type 1 (BpPV1), was detected in papillomas from a woylie (Bettongia penicillata ogilbyi). The circular, double-stranded DNA genome contains 7,737 bp and encodes 7 open reading frames (ORFs), E6, E7, E1, E2, E4, L2, and L1, in typical PV conformation. BpPV1 is a close-to-root PV with L1 and L2 ORFs most similar to European hedgehog PV and bandicoot papillomatosis carcinomatosis virus types 1 and 2 (BPCV1 and -2). It appears that the BPCVs arose by recombination between an ancient PV and an ancient polyomavirus more than 10 million years ago.

Hematologic and serum biochemical reference values for free-ranging northern hairy-nosed wombats.

Hematologic and serum biochemistry values were determined for 31 adult (21 male and 10 female) and four subadult male northern hairy-nosed wombats (Lasiorhinus krefftii) from the only existing population in Epping Forest National Park, Australia. Blood samples were obtained from free-ranging northern hairy-nosed wombats during trapping for population census and health and reproductive assessment in 1999. Hematologic and biochemical values were compared between adult males and adult females, and between adult and subadult wombats. Values were also compared with those previously published for southern hairy-nosed (Lasiorhinus latifrons) and common (Vombatus ursinus) wombats. The values from this study were used to create reference intervals, and they make up the first comprehensive hematologic and biochemical study for this highly endangered species.