Infectious Disease Surveillance in the Woylie (Bettongia penicillata).

Wild populations of the critically endangered woylie (Bettongia penicillata) recently declined by 90% in southwest Western Australia. Increased predation is the leading hypothesis for decline, but disease may be playing a role increasing susceptibility to predation. To explore this possibility, we surveyed woylie populations in the wild, in captivity and in a predator-free sanctuary for exposure to, and infection with, four known pathogens of macropods: herpesviruses, Wallal and Warrego orbiviruses, and Toxoplasma gondii. Our study found two of 68 individuals positive for neutralizing antibodies against known macropodid alphaherpesviruses. Three of 45 individuals were PCR positive for a herpesvirus that was shown to be a novel gammaherpesvirus or a new strain/variant of Potoroid Herpesvirus 1. Further sequence information is required to definitively determine its correct classification. There was no evidence of antibodies to orbivirus Wallal and Warrego serogroups, and all serological samples tested for T. gondii were negative. This is the first report of PCR and serological detection of herpesviruses in the woylie. Positive individuals did not demonstrate clinical signs of herpesviral diseases; therefore, the clinical significance of herpesviruses to wild woylie populations remains unclear. Further monitoring for herpesvirus infections will be important to inform disease risk analysis for this virus and determine temporal trends in herpesvirus activity that may relate to population health and conservation outcomes.

A NEED FOR DYNAMIC HEMATOLOGY AND SERUM BIOCHEMISTRY REFERENCE TOOLS: NOVEL USE OF SINE WAVE FUNCTIONS TO PRODUCE SEASONALLY VARYING REFERENCE CURVES IN PLATYPUSES (ORNITHORHYNCHUS ANATINUS).

Seasonal changes in hematology and serum biochemistry results, described by separate reference intervals for different seasons, have been reported in many animals. We developed a novel method to investigate seasonal variation in values and a reference tool (the reference curve) based on sine wave functions that, for suitable variables, represents data more appropriately than a fixed reference interval. We applied these techniques to values observed in blood samples from 126 adult wild platypuses ( Ornithorhynchus anatinus ; 58 females and 68 males). Samples were collected under isoflurane anesthesia from animals captured in the Inglis Catchment in northwest Tasmania. In general, packed cell volume (PCV), red cell count (RCC), and hemoglobin (Hb) values appeared to be lower than those in two studies that previously reported platypus hematology reference intervals. This likely resulted from reduced stress-related splenic contraction or isoflurane-associated splenic sequestration of red blood cells in our study. Reference curves were described for five variables (PCV, RCC, Hb, albumin, and magnesium). We found evidence that this seasonal variation may result from metabolic changes associated with seasonal variations in environmental temperature. These observations suggest that it is important for researchers reporting platypus hematology and serum biochemistry to look for seasonal changes in their data to ensure it is appropriately interpreted.

INVESTIGATION INTO INDIVIDUAL HEALTH AND EXPOSURE TO INFECTIOUS AGENTS OF PLATYPUSES (ORNITHORHYNCHUS ANATINUS) IN TWO RIVER CATCHMENTS IN NORTHWEST TASMANIA.

Changes in the health of individuals within wildlife populations can be a cause or effect of population declines in wildlife species. Aspects of individual platypus ( Ornithorhynchus anatinus ) health have been reported. However, holistic studies investigating potential synergistic effects of both pathogens and environmental factors are needed to expand understanding of platypus individual health. We collected baseline data on the health of platypuses in two Tasmanian river catchments (including evidence of the potentially fatal fungal disease mucormycosis) and on individual, demographic, and geographic patterns associated with health data results. We examined 130 wild platypuses from the Inglis River Catchment and 24 platypuses from the Seabrook Creek Catchment in northwest Tasmania between 29 August 2011 and 31 August 2013. More than 90% of captured platypuses were infected with ticks, Theileria spp., and trypanosomes. Evidence of exposure to other infections, including Salmonella spp., Leptospira spp., and intestinal parasites, was low (<10%). Three platypuses had single fungal granulomas in the webbing of a forefoot, but no evidence of mucormycosis was found in any of the study animals. Possible subclinical hepatopathies or cholangiohepatopathies were found in six platypuses. Exposure to infectious agents did not cluster geographically, demographically, or in individuals, and there was minimal evidence of morbidity resulting from infection. This study has provided important baseline data for monitoring the effects of threatening processes, including mucormycosis, on the health of infected populations.

Molecular characterisation and genetic variation of Elephant Endotheliotropic Herpesvirus infection in captive young Asian elephants in Thailand.

Elephant Endotheliotropic Herpesvirus (EEHV) is emerging as a new threat for elephant conservation, since being identified as the cause of severe, often fatal, haemorrhagic disease in young Asian elephants. To describe positive cases and the molecular relatedness of virus detected in elephants in Thailand, we re-examined all available of EEHV samples occurring in young elephants in Thailand between 2006 and 2014 (n=24). Results indicated 75% (18/24) of suspected cases were positive for EEHV by semi-nested PCR. Further gene analysis identified these positive cases as EEHV1A (72%, 13/18 cases), EEHV1B (11%, 2/18) and EEHV4 (17%, 3/18). This study is the first to phylogenetically analyse and provide an overview of most of the known EEHV cases that have occurred in Thailand. Positive individuals ranged in age from one to nine years, with no sex association detected, and occurred across geographical locations throughout the country. All individuals, except one, were captive-born. No history of direct contact among the cases was recorded, and this together with the fact that various subtype clusters of virus were found, implied that none of the positive cases were epidemiologically related. These results concur with the hypothesis that EEHV1 is likely to be an ancient endogenous pathogen in Asian elephants. It is recommended that active surveillance and routine monitoring for EEHV should be undertaken in all elephant range countries, to gain a better understanding of the epidemiology, transmission and prevention of this disease.

Isotopic Evidence of a Wide Spectrum of Feeding Strategies in Southern Hemisphere Humpback Whale Baleen Records.

Our current understanding of Southern hemisphere humpback whale (Megaptera novaeangliae) ecology assumes high-fidelity feeding on Antarctic krill in Antarctic waters during summer, followed by fasting during their annual migration to and from equatorial breeding grounds. An increase in the number of reported departures from this feeding/fasting model suggests that the current model may be oversimplified or, alternatively, undergoing contemporary change. Information about the feeding and fasting cycles of the two Australian breeding populations of humpback whales were obtained through stable isotope analysis of baleen plates from stranded adult individuals. Comparison of isotope profiles showed that individuals from the West Australian breeding population strongly adhered to the classical feeding model. By contrast, East Australian population individuals demonstrated greater heterogeneity in their feeding. On a spectrum from exclusive Antarctic feeding to exclusive feeding in temperate waters, three different strategies were assigned and discussed: classical feeders, supplemental feeders, and temperate zone feeders. Diversity in the inter-annual feeding strategies of humpback whales demonstrates the feeding plasticity of the species, but could also be indicative of changing dynamics within the Antarctic sea-ice ecosystem. This study presents the first investigation of trophodynamics in Southern hemisphere humpback whales derived from baleen plates, and further provides the first estimates of baleen plate elongation rates in the species.

Hematologic and Serum Biochemical Reference Intervals for the Bilby (Macrotis lagotis).

Hematologic and serum biochemistry blood reference intervals were established for the endangered marsupial, the bilby (Macrotis lagotis). Blood samples were collected from 135 bilbies that were housed in a captive breeding colony or were free ranging in a 10-ha exclosure. Statistical analyses were undertaken, and significant differences in parameters were found depending on age, sex, and captive or free-ranging status of animals. These reference intervals will assist ongoing health and conservation management of this endangered marsupial species.

Emerging infectious disease or evidence of endemicity? A multi-season study of beak and feather disease virus in wild red-crowned parakeets (Cyanoramphus novaezelandiae).

Beak and feather disease virus (BFDV) is a single-stranded DNA virus that is the etiological agent of beak and feather disease in both wild and captive parrots. Given that BFDV is globally recognized as a conservation threat for wild parrots, between 2011-2013, red-crowned parakeets (Cyanoramphus novaezelandiae, n = 229), which are endemic to New Zealand, were captured in mist nets on Tiritiri Matangi Island and Hauturu-o-Toi/Little Barrier Island (LBI), New Zealand, for disease surveillance. Blood and feathers from all birds were tested by PCR for BFDV, and full genomes were recovered and sequenced. A subset of blood samples (n = 96) were tested for antibodies to BFDV by the haemagglutination inhibition (HI) test. A further 238 feather samples were obtained from red-crowned parakeets from three sites in the Wellington region of the North Island, and these were screened for BFDV. The DNA-based prevalence of BFDV infection determined on Tiritiri Matangi Island was 1.09% (CI 95 %, 0.1-3.9%); on Hauturu-o-Toi/LBI, 4.4% (95% CI, 0.5%-15.1%); on Kapiti Island, 3.4% (CI 95%, 1.1-7.8%); at the ZEALANDIA-Karori sanctuary, 1.6% (95% CI, 0-8.4%); and on Matiu-Somes Island, 0% (CI 95%, 0-12.3%). Seroprevalence for BFDV, indicating prior or current exposure, in the Tiritiri Matangi Island population, it was 2% (CI 95%, 0-10.1%), and in the Hauturu-o-Toi/LBI population was 14% (CI 95%, 5.3-27.9%). BFDV-positive birds showed no signs of clinical disease, with the exception of an individual bird obtained opportunistically from Shakespear Regional Park during the study period, which had classical signs of feather loss. Phylogenetic analysis of the 11 full genome sequences recovered from BFDV-positive red-crowned parakeets revealed evidence of ongoing viral flow between red-crowned parakeets and eastern rosellas (Platycercus eximius) in the Hauraki Gulf/Auckland region, with separate but closely related strains from the Wellington region of the North Island. This is the first study to report HI results for a New Zealand endemic parrot species, and the first epidemiological analysis of serial cross-sectional surveys in a BFDV-infected population of red-crowned parakeets in New Zealand. We postulate that although BFDV remains a threat to small, isolated or naïve populations of parrots globally, the low viral prevalence in this and other studies suggests that native parakeets in New Zealand may act as dead-end or spillover hosts.

KNEMIDOKOPTINID (EPIDERMOPTIDAE: KNEMIDOKOPTINAE) MITE INFESTATION IN WILD RED-CROWNED PARAKEETS (CYANORAMPHUS NOVAEZELANDIAE): CORRELATIONS BETWEEN MACROSCOPIC AND MICROSCOPIC FINDINGS.

During a study on health and disease in Red-crowned Parakeets (Cyanoramphus novaezelandiae) on Tiritiri Matangi Island and Little Barrier Island (Hauturu-o-Toi) in New Zealand between 2011 and 2013, an outbreak of feather loss prompted the collection of skin biopsies (n = 135) under anesthesia from the head of captured birds. A subset of samples (n = 7) was frozen to obtain whole specimens for identification of ectoparasites. Mites (range 1-11) were observed in 79/135 (58.5%) skin biopsies, whereas feather loss was only found in 47/142 (33.1%) birds captured during the sampling period. Compact orthokeratotic hyperkeratosis and acanthosis were found in association with mites. Procnemidocoptes janssensi (Acari: Epidermoptidae, Knemidokoptinae) was identified from whole mites obtained from skin biopsies. We describe the presence, pathology, and stages of infestation for knemidokoptinid mange in a wild parrot population in New Zealand. Given the clinical and pathologic changes observed and poor knowledge of the parasite's New Zealand host and geographic distribution, further work is recommended for this and sympatric parrots, to understand relationships between the host, parasite, environment, and expression of disease. Results from this study reinforce the value of including biopsy samples for the investigation of skin disease in wild birds, particularly to link etiologic agents with pathologic changes.

Presentation and prognostic indicators for free-living black cockatoos (Calyptorhynchus spp.) admitted to an Australian zoo veterinary hospital over 10 years.

The veterinary records of three species of free-living, endangered black cockatoos (Calyptorhynchus spp.; n = 565) admitted to the Perth Zoo Veterinary Hospital in Western Australia during a 10-yr period (2000-09) were analyzed to determine the effect of clinical presentation and treatment on survival to release. The most-common reason for admission was trauma (at least 76.7% of cases), and trauma was also the most-frequent finding on necropsy examination (80.1% of cases). Anemia and paralysis-paresis were significant factors determining the decreased likelihood of survival of cockatoos undergoing rehabilitation. Human activities, in particular vehicle strike, were significant causes of morbidity and mortality in free-living black cockatoo populations.

Cetacean morbillivirus in coastal Indo-Pacific bottlenose dolphins, Western Australia.

Cetacean morbillivirus (CeMV) has caused several epizootics in multiple species of cetaceans globally and is an emerging disease among cetaceans in Australia. We detected CeMV in 2 stranded coastal Indo-Pacific bottlenose dolphins (Tursiops aduncus) in Western Australia. Preliminary phylogenetic data suggest that this virus variant is divergent from known strains.

Hematologic and plasma biochemical reference values for three species of black cockatoos (Calyptorhynchus species).

Black cockatoos in southwest Western Australia face population declines as a result of habitat loss and fragmentation, poaching, competition with other species, vehicle strikes, and shootings. In this study, hematologic and plasma biochemical reference values were determined for 3 endemic Western Australian black cockatoo species: Carnaby's cockatoo (Calyptorhynchus latirostris) (n = 34), Baudin's cockatoo (Calyptorhynchus baudinii) (n = 22), and forest red-tailed black cockatoo (Calyptorhynchus banksii naso) (n = 22). Significant sex-related differences were detected in Carnaby's cockatoos, and differences were also found among the 3 species. The forest red-tailed black cockatoos showed a markedly greater eosinophil count than did the 2 white-tailed species. Blood values from samples obtained by different collection methods differed significantly for Carnaby's cockatoos, which indicates that anesthesia may affect analyte values and that collection methods should remain consistent. The results of this study will be useful for clinically evaluating wild black cockatoos that are undergoing rehabilitation for release and will provide baseline data for future investigations of free-living black cockatoo health.