James Macgregor

Novel genotypes of Trypanosoma binneyi from wild platypuses (Ornithorhynchus anatinus) and identification of a leech as a potential vector

Little is known about the prevalence and pathogenesis of trypanosomes in Australian monotremes, and few genetic characterisation studies have been conducted with these haemoparasites. During the present investigation, molecular and microscopic methods were used to screen peripheral blood (n=28) and ectoparasites (n=10 adult ticks; n=5 tick nymphs; n=1 leech; and n>500 tick eggs) collected from wild Tasmanian platypuses (Ornithorhynchus anatinus), for the presence of trypanosomatid-specific DNA and/or trypomastigotes. The genes for the small ribosomal subunit RNA (18S rDNA) and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) were amplified and sequenced, prior to conducting phylogenetic analyses. The detection rate of the parasite-specific 18S rDNA in platypus blood was 85.7% (n=24/28), and the leech was also positive at both loci. Microscopically, high parasitaemia and the presence of abundant trypomastigotes, morphologically consistent with Trypanosoma binneyi Mackerras (1959), were observed in the blood films. Phylogenetic analyses at the 18S locus revealed the existence of four trypanosomatid-like genotypes, with variable similarity to two previously-described genotypes of T. binneyi (range of genetic p-distance: 0.0-0.5%). For the gGAPDH locus, for which only one T. binneyi sequence is available in GenBank, three genotypes closely related T. binneyi were identified (range of genetic p-distance: 0.1-0.4%). The leech-derived trypanosome isolate was virtually identical (at the two loci studied) to the other parasites sequenced from infected platypuses; however, the molecular or morphological identification of the leech species was not possible. Although further studies are required, the molecular detection of trypanosomes in an aquatic leech removed from a platypus, suggests the possibility that these haematophagous hirudineans may be a vector for T. binneyi (and closely related genotypes).

Preliminary investigation into the prevalence of mucormycosis in the platypus (Ornithorhynchus anatinus) in three catchments in north-west Tasmania

OBJECTIVE To investigate the distribution and prevalence of mucormycosis in platypus (Ornithorhynchus anatinus) from the Inglis, Emu and Black-Detention catchment areas in north-west Tasmania. PROCEDURE A field study was performed and resulted in the examination of 44 wild platypuses; in addition, one dead platypus and two live platypuses were examined after they were independently submitted to a local veterinary clinic. RESULTS No cases of mucormycosis were conclusively diagnosed. One platypus with signs consistent with those previously described in cases of mucormycosis was captured in the Emu River catchment. However, laboratory tests did not provide a definitive diagnosis for the lesion. Two platypuses from the Inglis catchment area had signs very similar to those previously described in cases of mucormycosis, but laboratory tests found Corynebacterium ulcerans to be the likely cause of the cutaneous ulcers on one of these platypuses and an unidentified fungal agent to be the cause of a cutaneous nodule in the other. CONCLUSIONS These findings do not prove that mucormycosis is absent from the populations studied. However, they may indicate that the prevalence of disease is low. The possibility that Mucor amphibiorum is present in a subclinical form in platypuses, or infecting another reservoir, is not excluded. The findings also suggest that caution should be exercised when diagnosing mucormycosis based on clinical findings alone and raise the possibility that some cases may have been incorrectly diagnosed.

Investigation into the characteristics, triggers and mechanism of apnoea and bradycardia in the anaesthetized platypus (Ornithorhynchus anatinus)

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’.

Investigation into individual health and exposure to infectious agents of platypuses (Ornithorhynchus anatinus) in two river catchments in northwest Tasmania

Abstract 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.

Novel use of in-stream microchip readers to monitor wild platypuses

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.

Dermatophilus congolensis Infection in Platypus (Ornithorhynchus anatinus), Tasmania, Australia, 2015

Abstract We report disease due to Dermatophilus congolensis infection in three of 13 (23%) platypuses (Ornithorhynchus anatinus) from a catchment in Tasmania, Australia. This pathogen has not previously been reported in platypuses. Two of the three infected platypuses had extensive scab formations, but no substantial hair loss was apparent.

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)

Abstract 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.

Assessing body condition in the platypus (Ornithorhynchus anatinus): a comparison of new and old methods

Abstract. 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.

Corrigendum to: Novel use of in-stream microchip readers to monitor wild platypuses

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.