Kristin Warren

The woylie or brush-tailed bettong (Bettongia penicillata) is critically endangered having suffered recent significant population declines in the wild. This project contributes to investigations aiming to determine if disease is a significant factor in woylie declines and lack of recovery. Surveys, examinations and sampling have been conducted over two years in three varying population management systems: a wild population in SW Western Australia; a nearby insurance population in the free-range predator-proof Perup Sanctuary; and a captive insurance population previously at Perth Zoo. Studying the wild-caught Sanctuary population has allowed the investigations to focus on the role of disease in the absence of introduced predators. Health testing has included haematology, biochemistry, and ecto and endoparasites, as well as more novel indicators of health such as anti-oxidant levels and the determination of stress levels using hair, faecal and serum glucocorticoids. The project with collaborators has validated the use of faecal glucocorticoids as a measure of stress for this species, and plans to also validate the use of hair. • Screening for significant marsupial pathogens includes toxoplasmosis and selected viruses including Herpes virus and Warrego & Wallal orbiviruses. Complete testing of samples and full data analyses are pending with the aim being to reveal differences in health, disease and stress levels between the three populations, and in doing so, highlight significant health-related causal factors of woylie declines and lack of recovery in the wild. Findings could aid in the recovery and management of this critically endangered species, as well as other free-ranging and captive marsupial populations. The presentation will focus on one research area of the project.

Introduced mammalian predators and anthropogenic habitat modification have resulted in the loss of 62 avian species since humans arrived in New Zealand. The conservation paradigm for New Zealand relies upon predator free, ecologically intact or restored islands to serve as sanctuaries and breeding sites for a range of threatened species. Island breeding programs and translocations to restore native fauna to the mainland and other off-shore sites, introduce specific risks related to disease spread. Beak and Feather Disease Vims (BFDV), was found in clinically affected juvenile Red Crowned Parakeet (RCP, Cyanoramphus novaezelandiae) on Hautum Island in 2008. This finding had major implications for conservation management of parrots in New Zealand, including the critically endangered kakapo (Strigops habroptilus) and orange fronted parakeet (Cyanoramphus malherbi). Feather loss had also been reported in RCP on nearby Tiritiri Matangi Island, suspected to be caused by BFDV. BFDV is endemic in the exotic Eastern rosella (Platycercus eximius) and Sulphur-crested cockatoo (Cacatua galerita) on the North Island of New Zealand. A spill over event likely spread the vims to RCP on nearby islands in the Hauraki Gulf of the Auckland region. Four cross-sectional surveys were conducted between 201 1-2012 on Tiritiri Matangi Island to determine the presence, prevalence and seasonal trends of key diseases of parrots that may result in feather loss. A total of 184 birds were captured in mist nets. All birds were anaesthetised to collect samples of blood, feathers and faeces, and in 2012 a skin biopsy from the head was included. Individuals were examined for general condition including stages of moult and body weight, and morphometrics and standardised photos were obtained. Hippoboscid flies were collected opportunistically when observed on birds. Blood and feather samples were tested for BFDV by PCR, and a subset of samples was tested for the presence of antibodies to BFDV using the haemagglutination inhibition test. Skin was processed routinely for histopathology and examined under light microscopy for evidence of BFDV as well as other causes of feather loss. The prevalence of feather loss in the study population changed substantially during the 2 year project, from 0% (95%CI: 0-6.7%) in April 2011, up to 45% (95%CI: 32.0-58.5%) in September 2012. This feather loss was predominantly around the head and neck region, with varying degrees of hyperkeratosis and lichenification evident on physical exam. BFDV was detected only in the first sampling session at 4% prevalence (95%CI: 0.5-13.0%), and was not correlated with clinical signs. A mite was detected in skin biopsies of all birds showing signs of feather loss and, increasingly, in skin biopsies from birds not showing changes to feathering. Examination of hippoboscid flies revealed female mites and eggs on the abdomen. The characteristic feather loss associated with this mite has been observed in other populations of RCP, as well as in several populations of the orange fronted parakeet. Continued monitoring of the study population is recommended to broaden our understanding of the factors that underpin the host-parasite relationship, and produce survival data to infer the impacts of this parasite on viability at individual and population scales.

Tiritiri Matangi Island in the Hauraki Gulf region of Auckland is an open sanctuary with a reintroduced population of kakariki (red crowned parakeets, Cyanoramphus novaezelandiae). These birds originally came from captivity in the 1970's. For the past decade, there have been increasing reports of feather loss in this species on the island. In 2008, beak and feather disease virus (BFDV) was detected nearby on Hauturu island, and therefore it was suspected the feather loss might have been due to a spread of this pathogen. However, a 2-year study into health and disease of red crowned parakeets on Tiritiri Matangi has revealed a different source of the clinical signs. During four cross sectional studies from 2011-12, feather loss increased from 8% (95%CI:2.3%-18.8%) up to 52% (95%(1:38.6%-65.2%) of the population. Skin biopsies from all birds in the second year of the study found a skin mite associated with thickening of the skin (acanthosis) and excessive keratin production (hyperkeratosis). Mites were also found in asymptomatic birds, suggesting a subclinical or carrier status. Whole mites were cleared in Hoyer's medium, with larval and female forms of Procnemidocoptes jansseni identified. This mite has only been formerly described in a lovebird from Zambia (Fain 1966), raising questions as to how it came to be present in wild kakariki in New Zealand. Another skin mite Hemimyialges macdonaldi, also reported to cause mange, was found in low numbers from skin as well as on hippoboscid flies removed from several birds. The relationship between the cosmopolitan H.macdonaldi, the dominant mite P.jansseni, and the clinical signs of mange requires further investigation. New Zealand is host to a unique avifauna, which has been significantly affected by the combined impacts of habitat modification and introduced mammalian predators. Many now thrive only on offshore islands or predator-free mainland sanctuaries, with ongoing conservation efforts reliant on re-introductions and translocations between these sites. These assisted movements, which may include periods of captive management, introduce specific biosecurity risks and potential for artificial spread of pathogens and parasites. Historically these activities took place in the absence of extensive or strategic disease screening, or prior to our current understanding of, or capacity to detect, key diseases. Results from this study will feed into captive and wild management of kakariki, specifically identifying new risks for translocations and re-introduction programs. The study also highlights the importance of epidemiological approaches to studying disease syndromes in wild populations.

Cetacean Morbillivirus (CMV) has caused several epizootics in cetaceans globally. An unusual mortality event among Tursiops aduncus in Perth, Western Australia (WA) coincided with increased strandings statewide in 2009. Necropsy of two dolphins identified mycotic encephalitis and pneumonia as the cause of death, respectively. Morbilliviral antigen was detected in multiple tissues using immunohistochemistry and confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) using primers for morbilliviral N and P genes. Sequencing of amplified RT-PCR products (approximately 238 bp in length from the N gene & 425 bp from the P gene) confirmed the presence of CMV in both dolphins and facilitated phylogenetic analysis. The N and P genes from the WA dolphins had respectively, 79% - 83% and 75% - 79% nucleotide identity to the highly conserved N and P genes of various strains of CMV. There was only 79% - 83% identity between the N and P nucleotide sequences from the WA dolphins and a variant from a Tursiops truncatus that stranded in Queensland (QLD), Australia in 2010. This is the first report of CMV-related mortality for T. aduncus in Australian waters and for the Indian Ocean. Preliminary phylogenetic data suggest that the WA variant is distinct from all other morbilliviruses, and is closely related to, but divergent from, other cetacean morbilliviruses. It is distinct from the QLD CMV variant, which is more closely related to the CMV isolates from the Northern Hemisphere. Furthermore, the WA variant appears to be the most closely related marine mammal morbillivirus to the terrestrial genus members identified to date. Evidence of a divergent CMV has implications for understanding the evolutionary history of CMV and supports ongoing surveillance and archival sampling to elucidate the role of CMV and other pathogens in future mortalities.

Piroplasms belonging to genera Theileria and Babesia (Phylum: Apicomplexa) are vector-borne protozoan haemoparasites with similar phenotype that infect erythrocytes of domesticated mammals and wildlife including birds. In addition, piroplasms of Theileria have an exoerythrocytic life cycle stage within the host’s white blood cells. Observations of intraerythrocytic piroplasms have been made sporadically in peripheral blood films of Australian native mammals since the first report was made nearly a Century ago and the advent of PCR has stimulated renewed research interest and descriptions of the molecular phylogeny of these organisms in a variety of marsupial hosts. A piroplasm (Theileria ornithorhynchi) has been described in previous studies of the platypus (Ornithorhynchus anatinus). As part of a study into the health and ecology of this iconic monotreme, blood samples were collected for haematological and biochemical analysis from wild-trapped platypuses in Tasmania. A subset of these samples, together with their ectoparasites, was evaluated for piroplasm infections by blood film microscopy and molecular analysis of the 18S ribosomal RNA gene, and phylogenetic analysis was performed with NJ trees using BIONJ. Estimates of evolutionary divergence between sequences were calculated by MEGA 5. A total of 27 blood samples and eight ectoparasites (seven ticks, one leech) were examined from 27 platypuses. All ticks were identified as Ixodes ornithorhynchi. Piroplasm infections were highly prevalent in the population studied (27/27; 100% infected); organisms were pleomorphic with infrequent tetrads and intra-leukocytic forms, thought to be Theileria schizonts, were occasionally observed. Anaemia was not detected. Many platypuses were co-infected with trypanosomes. All blood samples and three ectoparasites were positive for piroplasm DNA and phylogenetic analysis suggested the presence of more than one piroplasm species. This study emphasises the importance of combining traditional parasitological techniques (e.g. microscopy and tick examination) with molecular techniques if the life cycle of organism is to be understood.

Reptiles are commonly the subjects of biological or ecological research projects, and veterinarians or wildlife researchers may be required to euthanase a reptile if it sustains a severe injury associated with the research. When conducting euthanasia of any animal it is critical to confirm death. Whilst in mammals and birds euthanasia and confirmation of death can easily be accomplished, in reptiles these are not straight forward processes due to reptilian poikilothermic biology and physiology. Many traditional methods of reptile euthanasia are controversial and recommended methods of acceptable euthanasia vary amongst the different reptilian orders. Physical methods of euthanasia involving hypothermia or decapitation alone are considered inhumane and are not acceptable methods of euthanasia. Injectable pentobarbitone sodium is considered an acceptable method of euthanasia for all reptiles, except large crocodiles and other large reptile species where carcass removal in the wild may be problematic e.g. sea turtles. However, pentobarbitone sodium is a Scheduled 4 drug with requirements for storage in a locked environment and users other than registered veterinarians must apply for authorisation to administer scheduled drugs. Stunning and destruction of the brain is considered acceptable with reservations in some species of snakes and lizards. Humane euthanasia in reptiles is not easily accomplished and, whilst recognising limitations in accessing veterinary anaesthetic and euthanasia drugs, it can best be assured by using a two-stage euthanasia process – whereby the reptile is initially anaesthetised, and then euthanased by administration of pentobarbitone sodium or decapitation and brain destruction following anaesthesia.

Traditionally veterinary schools in Australasia have concentrated undergraduate training on the health and medicine of domesticated animals with an increasing emphasis on small animal medicine. However, veterinary practices are now called upon to treat and examine exotic pets including reptiles, amphibians, caged birds, native mammals, and rabbits and other small exotic pets. Historically, there has been little undergraduate training in these areas and veterinarians have had to learn from the literature, attend conferences and develop their interest and confidence to tackle these types of cases. Veterinary practices also increasingly see many injured or sick wildlife cases brought in by members of the public or by carers. Each practice in Australia treated on average 3.48 wildlife cases each week- at a time when there were 1792 practices in Australia. In 2003, across Australia each week practices were treating 6236 animals. Ten years later it is likely that this has increased considerably. Concurrently, there has been a marked increase in interest in emerging infectious diseases linked in many instances to wildlife such as SARS, Hendra virus, Nipah virus and this has led to new terms that have arisen such as ecohealth, ecosystem health and one health. In addition there have been diseases that have affected wildlife biodiversity eg chytriodiomycosis and beak and feather disease, and some of which often spillover in to domestic animals such as tuberculosis - these areas have led to the new term – Conservation medicine. Much has changed over the last 20 years but has undergraduate veterinary education adapted to these changes? Some veterinary schools in Australasia are starting to provide more training to support these emerging needs, ranging from individual sick and injured wildlife cases, to pet reptiles and birds, to diseases of wildlife populations which can affect biodiversity and the balance of ecosystems. What are the drivers of these changes? Should we be teaching veterinary students about the drivers of the changes in the health of ecosystems and biodiversity? This presentation provides a snapshot of the undergraduate training that is being provided in wildlife health in veterinary schools within Australasia by the analysis of data collected from a questionnaire sent to all schools. lt discusses how we can adapt the training in the future to ensure veterinary students are conversant with the challenges that lie ahead and can seek employment in these areas.

The IUCN Red List categorizes the Asian elephant as endangered, citing a drastic decline in the global population. Once widespread in Nepal, wild elephants are now limited to a few protected areas, and conservation of the species is of paramount concern. Decline in numbers predisposes this species to the impacts of such disease, through loss of genetic diversity, population fragmentation and increased interactions between humans, captive and wild populations. Of particular concern is the morbidity and mortality caused by tuberculosis (resulting from infection with Mycobacterium tuberculosis complex (MTBC) species) and endotheliotrophic elephant herpes vims (EEHV), both of which present a growing threat to the health and viability of Asian elephant populations worldwide. Tuberculosis is well documented in Nepal, and previous research has laid foundations for control and management of the disease. However, further research and recommendations are required, particularly to improve diagnostic techniques and identify and minimize disease risk factors. The Nepal Asian Elephant (Elephas maximus) Infectious Disease Mitigation and Prevention Initiative began in 2012 as a collaborative post-graduate research project, expanding into an ongoing initiative based at the Center for Molecular Dynamics Nepal (CMDN). Research and management is focused on Chitwan and Bardia National Parks, where the majority of Nepal's captive and wild populations reside. The initiative is endorsed by the Department of National Parks and Wildlife Conservation (Nepal), with technical support from the National Trust for Nature Conservation (NTNC) and Elephant Care International (ECI), and with academic expertise from Murdoch University, Tufts University, Johns Hopkins University, Columbia University and the Smithsonian Institute. The initiative aims to identify and minimize infectious disease risk factors at the captive-wild and captive-human interface, with particular focus on tuberculosis and EEHV in Nepal's captive and wild elephant populations. By controlling disease at the human-elephant and captive-wild interface, we seek to prevent transmission of disease to wildlife populations, and in the case of tuberculosis, to humans. Research aims to build upon existing studies through adopting a One Health approach to fill important knowledge gaps in disease prevalence, diagnosis and epidemiology, to identify management improvements, and to empower stakeholders through conservation education, community outreach and in-country capacity building. The purpose of this presentation is to update the research community and review the initiative's achievements over the past year, and to make recommendations for future directions to mitigate and prevent infectious diseases and facilitate a holistic conservation management plan for the preservation of the Asian elephant. It is hoped that findings may be applicable to other elephant range countries, and to zoological collections.

This poster presents preliminary findings from a proof of concept trial for satellite tracking of Baudin's cockatoos. The study follows the work by Christine Groom, which is successfully demonstrating that Carnaby's cockatoos can be tracked using satellite transmitters, and a trial which involved attaching transmitter devices to captive black cockatoos of all three species (Le Souef et al. 2013). Two rehabilitated adult female Baudin's cockatoos were anaesthetised and fitted with tail-mounted satellite trackers and released in Kelmscott in September 2012. Prior to release, both birds were health checked and flight tested to demonstrate fitness for release. The movements of the cockatoos were monitored according to Argos satellite transmissions, as well as ground truthing using flock sightings and reception of VHF signals from the satellite units using a radio telemetry antenna. According to their transmissions, the birds initially stayed in the Kelmscott area in close proximity to one another and other groups of Baudin's cockatoos in the area. However, after several days, one of the cockatoos flew south and joined a flock of Baudin's cockatoos migrating further south. Interestingly, during this migration, this bird returned to the Serpentine area from which she was originally found injured. This bird currently remains in the Beela area, 140km southeast of Perth, with other Baudin's cockatoos. The second cockatoo remained in the release area for several weeks before also moving south to Cardup, 33km southeast of Perth, where her transmitter was found two months later, still attached to the tail feathers which had possibly moulted out.