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Genetic diversity at loci concerned with fitness is an important part of the ability of a wild population to adapt to changes in its environment, including climatic events, disease and pollution. Research into the effects of genetic diversity on the impacts of disease on wildlife populations has focussed on genes of the major histocompatibility complex (MHC). This study investigated the genetic diversity at the MHC class II DZB gene, as well as the distribution of alleles of the same gene, for platypuses Ornithorhynchus anatinus in the Seabrook Creek Catchment in northwest Tasmania. This study detected 10 previously identified alleles and two previously unreported alleles at the MHC Class II DZB locus in 18 platypuses from the Seabrook Creek Catchment. An additional sequence isolated from two individuals was consistent with a pseudogene. Alleles were reasonably well distributed geographically through the catchment, but there was evidence of a degree of isolation at one site. Consistent with evidence that smaller wildlife populations have relatively low genetic diversity, and that there is relatively slow gene flow between river catchments, the observed genetic diversity at the MHC Class II locus was lower than those in larger previously studied river catchments but higher than those in two island populations. Consequently, this population of platypuses may have a limited capacity to respond to new infectious challenges, such as the fungal disease mucormycosis.

The platypus (Ornithorhynchus anatinus) is one of the world’s most evolutionarily distinct mammals, one of five extant species of egg-laying mammals, and the only living species within the family Ornithorhynchidae. Modern platypuses are endemic to eastern mainland Australia, Tasmania, and adjacent King Island, with a small introduced population on Kangaroo Island, South Australia, and are widely distributed in permanent river systems from tropical to alpine environments. Accumulating knowledge and technological advancements have provided insights into many aspects of its evolutionary history and biology but have also raised concern about significant knowledge gaps surrounding distribution, population sizes, and trends. The platypus’ distribution coincides with many of Australia’s major threatening processes, including highly regulated and disrupted rivers, intensive habitat destruction, and fragmentation, and they were extensively hunted for their fur until the early 20th century. Emerging evidence of local population declines and extinctions identifies that ecological thresholds have been crossed in some populations and, if threats are not addressed, the species will continue to decline. In 2016, the IUCN Red Listing for the platypus was elevated to “Near Threatened,” but the platypus remains unlisted on threatened species schedules of any Australian state, apart from South Australia, or nationally. In this synthesis, we review the evolutionary history, genetics, biology, and ecology of this extraordinary mammal and highlight prevailing threats. We also outline future research directions and challenges that need to be met to help conserve the species.

Consumption of poultry products contaminated with Salmonella is one of the major causes of foodborne diseases worldwide and therefore detection and differentiation of Salmonella spp. in poultry is important. In this study, oligonucleotide primers were designed from hemD gene and a PCR followed by high-resolution melt (HRM) curve analysis was developed for rapid differentiation of Salmonella isolates. Amplicons of 228 bp were generated from 16 different Salmonella reference strains and from 65 clinical field isolates mainly from poultry farms. HRM curve analysis of the amplicons differentiated Salmonella isolates and analysis of the nucleotide sequence of the amplicons from selected isolates revealed that each melting curve profile was related to a unique DNA sequence. The relationship between reference strains and tested specimens was also evaluated using a mathematical model without visual interpretation of HRM curves. In addition, the potential of the PCR-HRM curve analysis was evaluated for genotyping of additional Salmonella isolates from different avian species. The findings indicate that PCR followed by HRM curve analysis provides a rapid and robust technique for genotyping of Salmonella isolates to determine the serovar/serotype.

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.

Case report The clinical and laboratory findings in an orphaned juvenile female platypus (Ornithorhynchus anatinus) that presented with a severe anaemia and tick infestation are reported. The animal developed a terminal septicaemia and died. Antemortem clinical pathology, postmortem histopathology and 18S rDNA sequencing supported a diagnosis of extravascular haemolytic anaemia secondary to Theileria ornithorhynchi infection. Conclusion Although T. ornithorhynchi infection is common in the platypus, this is the first case in which it has been shown to cause a haemolytic anaemia in this species and molecular characterisation of the organism has been described. A review of the previous literature concerning T. ornithorhynchi and possible treatment options for future cases are discussed.

Chytridiomycosis in amphibians, and mucormycosis in the platypus Ornithorhynchus anatinus and amphibians, are serious fungal diseases affecting these aquatic taxa. In Tasmania, Australia, the fungi that cause these diseases overlap in range along with Phytophthora cinnamomi (Pc), an invasive fungal plant pathogen. To identify disinfectants that may be useful to reduce anthropogenic spread of these fungi to uninfected wilderness areas, for example by bush walkers and forestry or fire-fighting operations, we tested 3 disinfectants and a fire-fighting foam against Mucor amphibiorum (Ma) and tested 1 disinfectant and the foam against Batrachochytrium dendrobatidis (Bd). Combining the present study with previous work we found Bd was more susceptible to all 4 chemicals than Ma. Phytoclean, a disinfectant used at 2 to 10% for 30 s to control Pc, killed cultures of Bd at 0.075% and Ma at 5%, when also applied for 30 s. The disinfectant F10sc was not effective against Ma at standard exposures, but previous work shows Bd is killed at 0.03% with a 1 min exposure. Path-X is effective against Bd at 0.001% with a 30 s exposure and killed Ma at 1% with a 5 min exposure. Forexpan S, a foam added to water at 0.1 to 1% to control forest fires, killed Bd but not Ma when used at 1% for 2 min. Therefore, Phytoclean and Path-X have broader efficacy, although Path-X has not been trialled against Pc. Interestingly a positive mating strain of Ma (from a platypus) was more resistant to disinfectants than a negative strain (from a frog). Current protocols against Pc that involve high concentrations (10%) of Phytoclean are likely to reduce spread of pathogenic wildlife fungi, which is important for protecting biodiversity.

SUMMARY Campylobacter infection is a common cause of bacterial gastroenteritis in humans and remains a significant global public health issue. The capability of two multiplex PCR (mPCR)–high-resolution melt (HRM) curve analysis methods (i.e., mPCR1-HRM and mPCR2-HRM) to detect and differentiate 24 poultry isolates and three reference strains of Campylobacter jejuni and Campylobacter coli was investigated. Campylobacter jejuni and C. coli were successfully differentiated in both assays, but the differentiation power of mPCR2-HRM targeting the cadF gene was found superior to that of mPCR1-HRM targeting the gpsA gene or a hypothetical protein gene. However, higher intraspecies variation within C. coli and C. jejuni isolates was detected in mPCR1-HRM when compared with mPCR2-HRM. Both assays were rapid and required minimum interpretation skills for discrimination between and within Campylobacter species when using HRM curve analysis software.

To examine the infective dose, incubation period and disease progression of an isolate of Salmonella enterica serotype Typhimurium definitive type 160 (DT160) originating from a naturally-infected house sparrow (Passer domesticus) during an outbreak of the disease in New Zealand.Thirty-six house sparrows captured from the wild and free of Salmonella spp were divided into six groups of six birds, housed individually, and inoculated orally with phosphate buffered saline (PBS) or 10(1), 10(2), 10(3), 10(5), 2 x 10(8) colony forming units (cfu) of the outbreak strain of S. Typhimurium DT160. The birds were observed for 10 days for clinical signs and/or mortality, and faecal samples were collected to determine excretion of S. Typhimurium. The birds were euthanised 11 days post-inoculation (p.i.) and a wide range of tissue samples were collected for histopathological examination, and culture and typing of Salmonella spp. Macro-restriction profiling by pulsed-field gel electrophoresis (PFGE) using XbaI was performed for the epidemiological typing of S. Typhimurium DT160 isolates.Mortality in house sparrows inoculated with S. Typhimurium DT160 was dose-dependent, and 2/6 birds inoculated with 10(5) cfu and all six birds inoculated with 2 x 10(8) cfu died during the study. Infected sparrows displayed few clinical signs, apart from diarrhoea and/or polyuria, fluffed plumage, and sitting on the floor of the cage. Faecal excretion of DT160 occurred briefly in two birds inoculated with 10(2) cfu and four birds inoculated with 10(3) cfu, on most days in five birds inoculated with 10(5) cfu, and continuously in six birds inoculated with 2 x 10(8) cfu. DT160 was isolated from the livers of three birds which received 10(3) cfu, five birds dosed with 10(5) cfu, and all six birds given 2 x 10(8) cfu. Following necropsy, histopathological lesions similar to those seen in the natural disease were observed in the liver or spleen of three birds which received 10(3) cfu, and all birds dosed withor =10(5) cfu.The results indicate that an isolate of S. Typhimurium DT60 originating from house sparrows in New Zealand is pathogenic to these birds and that the response is dose dependent. The persistence and excretion of the pathogen may last for at least 10 days. This confirms that sparrows infected with DT160 could be a source of infection to humans and other in-contact animals.

Cryptococcus neoformans var. gattii has been shown to have a strong association with eucalypts frequently used by koalas and, not surprisingly, it has been shown to colonize the nasal cavities of koalas. The progression from nasal colonization to tissue invasion is critical to understanding the pathogenesis of cryptococcosis in this species and provides a model for pathogenesis of cryptococcosis in other species. Cryptococcal antigenaemia was detected in twenty-eight healthy koalas from three different regions. This was interpreted as representing limited subclinical disease. One koala developed cryptococcal pneumonia 6 months after leaving the study, whereas another developed cryptococcal meningoencephalitis during the course of the study. Opportunistic necropsies on ten antigen-positive koalas resulted in discovery of small cryptococcal lesions in two (paranasal sinus and lung, respectively). Our data suggest that cryptococcal antigenaemia occurs commonly in koalas, especially in areas with a high environmental presence of C n. var. gattii. Subclinical disease appears most likely to manifest as a small focal lesion in the respiratory tract. Possible outcomes include elimination by an effective immune response, quiescence with possibility of later re-activation or direct progression to overt disease. Symptomatic and subclinical cases showed differences in levels of antigenaemia. The data presented have significant implications for koalas in captivity.

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.

By relating respiratory and heart rate changes observed in some platypuses under isoflurane anaesthesia to those produced by various stimuli to the trigeminal nerve we propose that in platypuses the changes that can result from isoflurane gas in the nasal cavity are most appropriately described by the term ‘nasopharyngeal response’., Health and conservation research on platypuses (Ornithorhynchus anatinus) may require anaesthesia to reduce stress and the risk of injury to both the animal and the researcher, as well as to facilitate examination and sample collection. Platypus anaesthesia can be difficult to manage, with reports of periods of apnoea and bradycardia described. This study investigated the conditions around sudden-onset apnoea and bradycardia in 163 field-anaesthetized platypuses as part of a health study. Anaesthesia was induced and maintained using isoflurane delivered in oxygen by face mask. Sudden-onset apnoea and bradycardia was observed in 19% of platypuses, occurring either at induction of anaesthesia, during recovery, or both. At induction, occurrence was more often recorded for adults (P = 0.19) and was correlated with low body temperature (P

Nine male and five female adult free-living platypuses, obtained in a prospective capture-release study from northern Tasmania, exhibited gross features of cutaneous mycosis caused by Mucor amphibiorum. The lesions were present on the hind limbs (six cases), front limbs (four), tail (five), dorsal trunk (three) and ventral trunk (one). They varied in size, and ranged from raised red nodules or plaques, which sometimes exuded purulent material, to ulcerated lesions with central cavitation, red exuding centres and raised epidermal margins. Older lesions were covered either partly or fully by thickened and irregular epidermis. Histological examination of skin biopsies revealed discrete, poorly encapsulated granulomas, or more commonly a diffuse granulomatous or pyogranulomatous inflammation. Inflammatory cells consisted of neutrophils or eosinophils, sparse plasma cells and lymphocytes, many macrophages and occasional multinucleated giant cells. Fibrovascular tissue was diffusely and irregularly scattered in the granulomatous regions. Sphaerules characteristic of M. amphibiorum infection were observed in all lesions. The cutaneous distribution of the lesions and the natural history of the platypus indicated that entry of M. amphibiorum may have been via superficial skin wounds. T cells were the predominant infiltrating lymphoid cells in the diffuse lesions, indicating the importance of the cell-mediated response to infection.

Objective To determine whether there are haematological, serum biochemical and serological differences between platypuses (Ornithorhynchus anatinus) with and without granulomatous dermatitis due to Mucor amphibiorum infection. An additional objective was to establish reference haematological and serum biochemical ranges for the species in Tasmania. Design A clinicopathological and serological study. Animals A total of 37 free-living adult platypuses captured from streams and dams in Northern Tasmania were used in the clinicopathological study. Twenty-seven were clinically normal and 10 had mycotic granulomatous dermatitis. A total of 22 platypuses (20 adult and 2 juvenile) were used for the serosurvey. Eighteen were captured from streams in Northern Tasmania, and four were submitted for necropsy. Results Platypuses with mycotic ulcerative dermatitis had significantly smaller packed red cell volumes, haemoglobin concentrations, lymphocyte counts, serum cholesterol and calcium concentrations, and higher serum globulin and potassium concentrations than clinically normal animals. The lymphopenia and hyperkalaemia were thought to be clinically significant. Numbers of Trypanosoma binneyi in blood smears were similar between the two groups. Diseased platypuses had higher concentrations of serum antibody against Mucor amphibiorum as determined by ELISA compared to clinically normal platypuses. Conclusion Platypuses affected by mycotic granulomatous dermatitis showed haematological and serum biochemical changes when compared to clinically normal animals from the same Tasmanian sites. A serological survey may be a useful method for detecting the prevalence of exposure to Mucor amphibiorum and humoral immunity in platypus populations both in Tasmania and the mainland of Australia.

The gross and histological appearance and the distribution of T and B lymphocytes and plasma cells are described for lymphoid tissues obtained from 15 platypuses. The spleen was bilobed and surrounded by a thick capsule of collagen, elastic fibres and little smooth muscle. White pulp was prominent and included germinal centres and periarterial lymphoid sheaths. Red pulp contained haematopoietic tissue. A thin lobulated thymus was located within the mediastinum overlying the heart. The cortex of lobules consisted of dense aggregates of small and medium lymphocytes, scattered macrophages and few reticular epithelial cells. In the medulla, Hassall's corpuscles were numerous, lymphocytes were small and less abundant, and reticular cells were more abundant than in the cortex. Lymphoid nodules scattered throughout loose connective tissue in cervical, pharyngeal, thoracic, mesenteric and pelvic sites measured 790 +/- 370 microm (mean +/- S.D., n = 39) in diameter, the larger of which could be observed macroscopically. These consisted of single primary or secondary follicles supported by a framework of reticular fibres. Macrophages were common in the germinal centres. The platypus had a full range of gut-associated lymphoid tissue. No tonsils were observed macroscopically but histologically they consisted of submucosal follicles and intraepithelial lymphocytes. Peyer's patches were not observed macroscopically but histologically they consisted of several prominent submucosal secondary follicles in the antimesenteric wall of the intestine. Caecal lymphoid tissue consisted of numerous secondary follicles in the submucosa and densely packed lymphocytes in the lamina propria. Bronchus-associated lymphoid tissue was not observed macroscopically but was identified in 7 of 11 platypus lungs assessed histologically. Lymphoid cells were present as primary follicles associated with bronchi, as aggregates adjacent to blood vessels and as intraepithelial lymphocytes. The distribution of T lymphocytes, identified with antihuman CD3 and CD5, and B lymphocytes and plasma cells, identified with antihuman CD79a and CD79b and antiplatypus immunoglobulin, within lymphoid tissues in the platypus was similar to that described in therian mammals except for an apparent relative paucity of B lymphocytes. This study establishes that the platypus has a well-developed lymphoid system which is comparable in histological structure to that in therian mammals. It also confirms the distinctiveness of its peripheral lymphoid tissue, namely lymphoid nodules. Platypus lymphoid tissue has all the essential cell types, namely T and B lymphocytes and plasma cells, to mount an effective immune response against foreign antigens.

Objective Correlation of immunophenotype with history, anatomical and morphological features of lymphoid neoplasia in the koala. Methods Routine necropsies were performed on 51 koalas with suspected lymphoid neoplasia between 1986 and 1997 in New South Wales and Queensland. Immuno-phenotyping was by an immunoperoxidase method utilising species cross-reactive antibodies raised against human lymphocytes and an antibody raised against koala IgG. Cases were classified according to organs and tissues affected and the morphological features of neoplastic cells. Results Twenty-six (51%) of the cases were of the T cell immunophenotype, 12 (24%) were of B cell immunopheno-type and 13 (25%) did not stain. The age and sex of koalas did not correlate with immunophenotype (P = 0.686 and P = 1.000, respectively). Thirty-two cases were leukaemic and 36 had multiple organ involvement, probably reflecting presenta tion of koalas at advanced stages of disease. Abdominal tissue involvement was most common (44 cases), followed by nodal (32), atypical (21) and cervicomediastinal (14). The T cell immunophenotype was over-represented among the leukaemic cases (P = 0.013). Generally, the T cell immunophenotype predominated except for many affected atypical tissues. Neoplastic cells were mostly of medium nuclear size with round to oval nuclei. No correlations were found for cell morphology, mitotic index and immunopheno-type. Conclusion The prognostic value of an immunopheno-typic, anatomical and morphological basis for the classifica tion of lymphoid neoplasia in the koala currently is limited by the need to detect these neoplasms at an early age, the requirement for freshly fixed tissues and the restricted range of available cross-reacting antibodies.

Body condition is an important aspect of the health of any animal. The current standard method of body condition assessment in the platypus is the tail volume index (TVI). Although the tail is the largest repository of fat in the platypus, the reliability of TVI has not been adequately demonstrated. The aims of this study were, first to assess performance of the TVI, and second, to develop and evaluate performance of new techniques for routine field assessment of platypus body condition. Morphometric data were collected under anaesthesia from 137 adult wild platypuses (74 males, 63 females) captured in north-west Tasmania; ultrasound images of tail fat were also collected from 100 of these individuals (54 males, 46 females). Three new indices for platypus body condition were identified. An objective tail fat index (Relative Tail Fat Volume: RTFVTBL) was developed, based on cross-sectional area measurements taken from detailed ultrasound images compared with total body length (TBL). Two body condition indices intended for routine field use were developed – one based on body mass (mb) and bill width (BW) (Body Condition Index; BCIBW), and the other based on a single linear ultrasonographic measurement of tail fat depth and BW (Relative Fat Depth; RFDBW). Results indicated that RFDBW outperforms TVI as an index of platypus tail fat. Further work, however, is needed to determine the relationship between tail fat and total body fat in the platypus before conclusions can be drawn about the effectiveness of BCIBW as a body condition index.

Little is known of the current status of platypus (Ornithorhynchus anatinus) populations in the Murrumbidgee catchment and other west-flowing rivers in the Murray–Darling Basin. Platypus distribution in the Murrumbidgee catchment was determined from sightings, reports by government departments, the literature and a capture–release study. The platypus was found to be widespread in the catchment, including all subcatchment and elevation categories, but most reports were from the middle and upper subcatchments. Sixty-five captures of 55 individual platypuses were made during 61 trap-nights to 31 study sites in the Murrumbidgee catchment during 2009–10. Only three juveniles (all females) were captured and recapture rates were low, with only nine platypuses captured more than once. Adult males had body weights of 980–2180 g and body lengths of 42–60 cm (n = 32). Adult females weighed 795–1480 g and were 38–52 cm long (n = 20). Tail fat index was significantly affected by season and platypus age, while in comparison, season, platypus length and weight had a significant effect on tail volume index. This study established a baseline that provides a starting point from which the impacts of environmental disturbances or diseases occurring over time can be measured and investigated.

The kakapo (Strigops habroptilus) is a critically endangered New Zealand parrot. The presence of house sparrows (Passer domesticus) infected with a potentially pathogenic phage type of Salmonella enterica on the 2 islands that contain the remaining kakapo caused concern that this phage type might infect the kakapo and threaten their survival. This study sought to determine the presence or absence of Salmonella and Campylobacter species in the kakapo population of Codfish Island, New Zealand. Fecal and cloacal swab samples from kakapo were collected during an annual health examination and cultured for Salmonella and Campylobacter species. None of the cultures from the 39 birds sampled were positive for either organism. Results of this study suggest that kakapo on Codfish Island are not currently infected with these organisms. However, management changes have been made to reduce the risk of disease transmission from feral birds, and surveillance for these organisms in the kakapo will be continued.

Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates.

A variety of techniques have been used to monitor platypus populations to assess the impacts of the threats they face, but each technique has limitations. In this study we investigated the novel use of in-stream microchip readers, to remotely monitor the movements of microchipped wild platypuses. Over 13 months, we recorded movements of 18 microchipped individuals past nine fixed locations in the Inglis Catchment in northwest Tasmania, using three units of which all were capable of detecting Trovan® unique microchips and two were additionally capable of detecting ISO microchips. Each site was monitored one or two times, for durations of 8–39 days. We undertook direction of movement investigations during two monitoring periods, by placing the antennas from two systems in the same creek within 3 m of each other. In a total of 264 days of monitoring, 528 platypus observations were made from 18 individual platypuses, consisting of 13 of 18 (72%) platypuses captured at the monitoring sites within 16 months prior to monitoring, two platypuses captured at other sites in the same time period, and three of seven (43%) individuals microchipped 3–5 years previously. This number of platypus observations, in combination with the stable number of platypuses observed per day, the range of movement behaviours recorded and the results of the direction of movement investigations, indicates that at appropriate sites, in-stream microchip readers are an effective method of monitoring the movements and survivorship of microchipped wild platypuses.

Mucormycosis in the platypus and the anuran (frogs and toads) is a serious fungal disease affecting these aquatic taxa. Mucor amphibiorum infection causes significantmorbidity and mortality in free-living platypuses in Tasmania. Infection has also been reported in free-ranging cane toads and frogs from mainland Australia, but not confirmed in platypuses from themainland. This paper reviewsmucormycosis in the platypus and anuran, including consideration of the clinical, epidemiological, pathological and diagnostic features.

This study reports an outbreak of salmonellosis due to S. Typhimurium DT160 which caused extensive mortality in wild birds and enteric disease in humans in New Zealand during the winter and spring months of the year 2000.Necropsies were performed and microbiological examinations undertaken on wild birds from populations in which mass mortality was reported, and on captive indigenous birds which died suddenly during the winter and spring of 2000. Affected tissues were examined histologically and isolates of S. Typhimurium were phage typed and examined using pulsedfield gel electrophoresis (PFGE). Isolates of S. Typhimurium obtained from cases of human enteric disease which occurred during these months were phage typed, examined using PFGE and compared with the bird isolates.Central and northern areas of the South Island and the southern North Island were worst affected with die-offs of several hundreds of sparrows and other birds reported in rural areas. Mortalities reached a peak in winter (July-August) 2000 and decreased to small numbers during the spring and early summer. The birds usually died of an acute septicaemia with multifocal necrotising lesions in the liver and spleen. Human cases throughout the country increased gradually over the same period. Isolates from birds, livestock and humans examined using PFGE were indistinguishable from one another.This strain of Salmonellahas emerged as a major cause of septicaemia in wild birds in New Zealand. Because of the close association between house sparrows (Passer domesticus) and humans, the organism also poses a serious zoonotic risk. The possibility that the infection may spread to involve indigenous species needs investigation.

To investigate the role of free-living animals such as sparrows, rodents and flies as potential reservoirs of Campylobacter spp on a dairy farm, and to assess the genetic diversity among Campylobacter isolates from the farm and an urban source.A total of 290 samples (bovine, passerine and rodent faeces, and whole flies) were collected from a large commercial dairy farm in the Manawatu district in New Zealand, and from faeces from urban sparrows in a nearby city. Other samples collected from the dairy farm included five from silage, two from aprons worn by workers during milking, two from workers' boots and two from water in troughs in a paddock. Isolates of thermophilic Campylobacter spp were identified morphologically and phenotypically and further characterised molecularly using pulsed-field gel electrophoresis (PFGE) and the restriction enzyme SmaI.Campylobacter jejuni was the only Campylobacter species isolated from all samples. The highest prevalence was found in faeces from dairy cows (54%), followed by faeces from sparrows from the urban area (40%) and the farm (38%), and from rodents (11%) and whole flies (9%). Other samples from the farm environment such as silage, trough water, and workers' aprons and boots were also positive for C. jejuni. Of the 22 restriction patterns obtained, seven were common to more than one source.Cattle, sparrows, rodents and flies are potential reservoirs of C. jejuni on dairy farms. Identical clones of C. jejuni carried by cattle, sparrows, flies and rodents probably indicate a common source of infection. The high level of asymptomatic carriage of C. jejuni by healthy dairy cows could be sufficient to maintain infections within the dairy farm surroundings via environmental contamination.

A variety of techniques have been used to monitor platypus populations to assess the impacts of the threats they face, but each technique has limitations. In this study we investigated the novel use of in-stream microchip readers, to remotely monitor the movements of microchipped wild platypuses. Over 13 months, we recorded movements of 18 microchipped individuals past nine fixed locations in the Inglis Catchment in northwest Tasmania, using three units of which all were capable of detecting Trovan® unique microchips and two were additionally capable of detecting ISO microchips. Each site was monitored one or two times, for durations of 8–39 days. We undertook direction of movement investigations during two monitoring periods, by placing the antennas from two systems in the same creek within 3 m of each other. In a total of 264 days of monitoring, 528 platypus observations were made from 18 individual platypuses, consisting of 13 of 18 (72%) platypuses captured at the monitoring sites within 16 months prior to monitoring, two platypuses captured at other sites in the same time period, and three of seven (43%) individuals microchipped 3–5 years previously. This number of platypus observations, in combination with the stable number of platypuses observed per day, the range of movement behaviours recorded and the results of the direction of movement investigations, indicates that at appropriate sites, in-stream microchip readers are an effective method of monitoring the movements and survivorship of microchipped wild platypuses.

Over a 22-month period, sequential nasal and skin swabs were obtained from 52 healthy captive koalas (Phascolarctos cinereus) from the Sydney region. Cryptococcus neoformans was isolated in 17 koalas from 64 of 262 (24%) nasal swabs and from nine of 262 (3%) skin swabs. Prevalence of nasal colonization varied seasonally from 12% (3/25) to 38% (10/26). Cryptococcus neoformans var. gattii alone was cultured from 37, var. neoformans alone from 22 and both varieties from five nasal swabs. Of 33 koalas sampled on three or more occasions, organisms were isolated persistently from six, occasionally from eight and never from 19. Two koalas were persistently and heavily (>/=100 colonies/plate) colonized by C. neoformans var. gattii and two with var. neoformans. Isolation of C. neoformans var. gattii from the skin was low grade and sporadic. No koalas from which C. neoformans was persistently isolated showed clinical signs of cryptococcosis and all except one had a negative latex cryptococcal antigen test, therefore the nasal cavity was presumed to be colonized by, rather than infected with, C. neoformans. Preliminary observations of koalas from Coffs Harbour indicated a much higher prevalence of colonization by C. neoformans, suggesting that environmental factors influenced the extent of carriage by C. neoformans.

Objective To describe a case of diabetes mellitus in a koala (Phascolarctos cinereus). Design A case report with controls. Procedures We describe clinical and laboratory findings in a 6-year-old, free-living, female koala presented with traumatic injury and subsequently found to have polydipsia, hyperglycaemia and glucosuria. Over a 5 week period, serum biochemical analyses, haematological examinations, urinal-yses, measurement of serum insulin and fructosamine concentrations, necropsy, histopathological examination of a range of tissues and immunohistochemical examination of the pancreas for insulin-containing cells were done. For reference purposes, serum insulin and fructosamine concentrations were determined in four and two healthy koalas, respectively, and three healthy koalas pancreases were examined histo-logically and immunohistochemically. Results The koala had persistent hyperglycaemia, hyperlipidaemia, hyponatraemia, hypochloraemia and glucosuria. Serum insulin concentration of the diabetic koala was only marginally smaller than that of healthy koalas, but all concentrations were smaller than reference concentrations in dogs and people. Fructosamine concentration did not allow the diabetic koala to be distinguished from healthy koalas and concentrations of all koala analytes were greater than expected for healthy dogs and people. Histopathological examination revealed extensive degeneration of pancreatic islet cells and fatty infiltration of hepatocytes. Immunoperoxidase staining revealed decreased or absent insulin in the b cells of the affected koala. Conclusion Clinical signs, clinicopathological results and histopathological changes were consistent with diabetes mellitus. The pathogenesis of the condition could not be determined but may have been related to the administration of a parenteral corticosteroid preparation, the stress of capture or tissue damage and inflammation.

Many infectious agents and parasites have been reported from the platypus (Ornithorhynchus anatinus), but most do not cause serious disease. The fungus, Mucor amphibiorum, is the only disease agent known to cause significant morbidity and mortality in the free-living platypus in Tasmania. Infection has also been reported in free-ranging cane toads and green tree frogs from mainland Australia, but not confirmed in platypuses from the mainland. This paper reviews mucormycosis in the platypus and includes the epidemiology, clinical features, mycology, pathology as well as possible surveillance, treatment and/or control modalities. The emergence and geographical spread of mucormycosis as a disease entity in Tasmanian platypuses from 1982 till 2005 are discussed. The host, agent and environment factors of the traditional epidemiological paradigm are discussed as they contribute to the conditions that lead to the selection or emergence of Mucor amphibiorum as a pathogen in a population of platypuses.

Over a twelve month period, 25 platypuses (Ornithorhynchus anatinus) were presented for necropsy. The most common causes of mortality were dog predation (40%), road trauma (28%), starvation and/or exposure (16%) and Mucor-mycosis (8%). Mucor amphibiorum is the only disease agent known to cause significant morbidity and mortality in free-living O. anatinus in Tasmania. This fungus causes a severe, granulomatous, commonly ulcerative, skin condition in this species. A study was conducted to define the nature and extent of the granulomatous dermatitis caused by M. amphibiorum. The mycotic granulomatous dermatitis of O. anatinus appears to be confined to streams in the north and midlands. The prevalence of the disease at two infected sites was 36% (n=36) and 66% (n=3). Lesions included abscesses, ulcers, granulation tissue and nodules. Of the 17 diseased animals captured, II were adult male, 5 were adult female and 1 was a juvenile female. Thirteen isolates of M. amphibiorum were cultured from skin lesions, all were of the positive mating type. Mucor amphibiorum was not isolated from 40 faecal or 8 skin samples from O. anatinus. No M. amphibiorum was isolated from 14 environmental samples. Mucor circinelloides and M. saturninus were isolated from soil, and M. circinelloides and M. hiemalis were isolated from faecal samples from O. anatinus.

Mucor amphibiorum is the only pathogen known to cause significant morbidity and mortality in the free-living platypus (Ornithorhynchus anatinus) in Tasmania. Infection has also been reported in free-ranging cane toads (Bufo marinus) and green tree frogs (Litoria caerulea) from mainland Australia but has not been confirmed in platypuses from the mainland. To date, there has been little genotyping specifically conducted on M. amphibiorum. A collection of 21 Mucor isolates representing isolates from the platypus, frogs and toads, and environmental samples were obtained for genotypic analysis. Internal transcribed spacer (ITS) region sequencing and GenBank comparison confirmed the identity of most of the isolates. Representative isolates from infected platypuses formed a clade containing the reference isolates of M. amphibiorum from the Centraal Bureau voor Schimmelcultures repository. The M. amphibiorum isolates showed a close sequence identity with Mucor indicus and consisted of two haplotypes, differentiated by single nucleotide polymorphisms within the ITS1 and ITS2 regions. With the exception of isolate 96-4049, all isolates from platypuses were in one haplotype. Multilocus fingerprinting via the use of intersimple sequence repeats polymerase chain reaction identified 19 genotypes. Two major clusters were evident: 1) M. amphibiorum and Mucor racemosus; and 2) Mucor circinelloides, Mucor ramosissimus, and Mucor fragilis. Seven M. amphibiorum isolates from platypuses were present in two subclusters, with isolate 96-4053 appearing genetically distinct from all other isolates. Isolates classified as M. circinelloides by sequence analysis formed a separate subcluster, distinct from other Mucor spp. The combination of sequencing and multilocus fingerprinting has the potential to provide the tools for rapid identification of M. amphibiorum. Data presented on the diversity of the pathogen and further work in linking genetic diversity to functional diversity will provide critical information for its management in Tasmanian river systems.

A study was conducted to determine the geographic distribution, mean length and weight of the platypus (Ornithorhynchus anatinus) in Tasmania. The species was found to be common and widespread on the mainland of Tasmania and present on King Island. Mean total lengths of adult males and females were 57. 1±3.5 cm (n=44) and 47.9±2. 1 cm (n=29) respectively. Mean weights of adult males and females were 23 13±37 1 g (n=42) and 1337±206 g (n=29) respectively. Tasmanian platypuses were generally larger than those from the mainland.

The thymuses of 44 koalas, ranging from less than 30 d to more than 14 y of age, were examined histologically and immunohistologically. The thymuses from 17 of these koalas dying acutely through trauma were regarded as not being significantly affected by disease and formed the basis for study of the normal thymus. Most other koalas had chronic illness and, consequently, disease affected (involuted) thymuses. Histologically, thymuses showed obvious corticomedullary differentiation with small Hassall's corpuscles visible in koalas more than 8 mo of age. Most cortical and medullary lymphocytes stained for CD3 and CD5 (T lymphocyte markers) while some cells (predominantly medullary) stained for CD79b (B lymphocytes and plasma cells), IgG (plasma cells) or MHC class II (reticular epithelium, macrophages and possibly lymphocytes). Adults of up to 5-6 y of age which had died through trauma had little evidence of involution and had prominent Hassall's corpuscles and medullary epithelial thymocytes. Thymic eosinopoiesis was an inconsistent finding. In traumatised animals over this age, involution was obvious with fibrous replacement of lobules, loss of Hassall's corpuscles and the development of dilated ducts lined by nonciliated epithelium. However, loss of lymphocytes was gradual and pockets of lymphocytes, centrally located in lobules, were still present in the oldest koala examined. In these involuted thymuses, remaining lymphocytes stained for CD3 and lesser numbers of CD5 and CD79b. Plasma cells were common and often stained both for IgG and MHC class II. Thymuses of chronically diseased koalas showed accelerated involution when age matched with thymuses from traumatised koalas. Chronically ill koalas as young as 18-24 mo showed advanced involution, but the morphological and immunohistological characteristics of involuted thymus from diseased koalas could not be distinguished from those of involuted thymuses derived from traumatised koalas. It was concluded that development of the koala thymus is completed at 8 mo of age and that for normal koalas involution is a gradual process which starts not at but after sexual maturity. Immunohistological characterisation of the thymus was comparable to that reported for a variety of eutherian mammals.