CLINICAL PATHOLOGY OF CAPTIVE ENDANGERED MAHOGANY GLIDERS (PETAURUS GRACILIS) IN RESPONSE TO ENVIRONMENTAL CHANGE.

The impetus of this study was the imperative to establish blood biomarker values for clinically healthy mahogany gliders (Petaurus gracilis) in order to monitor the health status of eight captive individuals during their movement to a new facility. The study established ranges for 18 hematologic and 21 biochemical blood biomarkers for healthy individuals in a captive environment. The reported values are consistent with those published for other Australian glider and possum species. No statistically significant differences were found between the sexes, but significant age effects were observed. Specifically, subadult animals reported significantly higher total white cell counts, lymphocyte counts, alkaline phosphatase, creatine kinase, and glucose and chloride levels, compared to adult animals. Although there were no clinically significant changes in blood biomarkers associated with the relocation, many of the hematologic and biochemical biomarkers demonstrated the expected changes associated with the physiological stress of relocation. Specifically, triglycerides, glucose, globulins, creatinine kinase, aspartate transferase (AST), total protein, urea, phosphorous, potassium, sodium, chloride, neutrophils, and hematocrit showed changes with the large environmental change. The majority of the blood biomarkers returned to baseline levels 5 wk postrelocation, with all but one aged animal showing no signs of chronic health derangements following the relocation. The abnormal blood biomarker profiles of two geriatric individuals, one male diagnosed with pericloacal and adrenal gland tumors at the beginning of the study, and one female diagnosed with chronic urinary tract infections and suspected bone marrow disease following the relocation, are presented. The findings of this study inform the health monitoring of native gliders in captivity, rehabilitation, and clinical research scenarios. These findings also provide useful baseline data to aid in the health assessment of captive-bred individuals during their reintroduction into free-living populations.

SERUM BIOCHEMISTRY OF FREE-RANGING SOUTHERN HAIRY-NOSED WOMBATS (LASIORHINUS LATIFRONS).

Listed as near-threatened by the International Union for Conservation of Nature (IUCN), the southern hairy-nosed wombat (SHNW, Lasiorhinus latifrons) faces threats such as drought, habitat degradation and loss, disease, and persecution because of competition with agriculture. To assist with evaluation of wombat health, this study reports serum biochemical reference intervals (RIs) for wild-caught SHNW from South Australia established from 126 apparently healthy SHNW using a Beckman Coulter AU480® Automated Chemistry Analyzer using RefVal Advisor. Partitioning of RIs for male and female wombats and for the two methods of sampling was performed as appropriate, and additional significant differences (P < 0.05) in biochemical profiles were identified across age class and season examined. A number of differences were observed between male and female wombats, most notably higher creatinine, urea, and sodium in females. Subadult and juvenile wombats had significantly lower total protein, globulin, and ALT activity, and significantly higher ALP activity than adults. Wombats sampled in winter and spring had significantly higher total protein, albumin, potassium, bicarbonate, and enzyme activities (ALP, ALT, AST, GGT, GLDH, lipase), and significantly lower glucose and creatinine when compared to individuals sampled in summer and autumn. Differences in CK activity and anion gap observed between the two methods of sampling likely reflect delay and handling of animals between capture and blood collection. The serum biochemical RIs documented here are considered representative of a population of healthy SHNW, providing a tool for health assessment and monitoring of SHNW health in South Australia and elsewhere.

In vitro binding of cefovecin to plasma proteins in Australian marsupials and plasma concentrations of cefovecin following single subcutaneous administration to koalas (Phascolarctos cinereus).

BACKGROUND: Cefovecin has a long duration of antibiotic activity in cats and dogs, somewhat attributable to its high plasma protein binding. AIMS: To determine the cefovecin binding to plasma proteins in vitro in selected Australian marsupials and to quantify the change in cetovecin concentration over time following subcutaneous injection in koalas. METHODS AND RESULTS: Various cefovecin concentrations were incubated with plasma and quantified using HPLC. The median (range) bound percentages when 10 µg/mL of cefovecin was incubated with plasma were 11.1 (4.1-20.4) in the plasma of the Tasmanian devil, 12.7 (5.8-17.3) in the koala, 18.9 (14.6-38.0) in the eastern grey kangaroo, 16.9 (15.7-30.2) in the common brush-tailed possum, 37.6 (25.3-42.3) in the eastern ring-tailed possum and 36.4 (35.0-38.3) in the red kangaroo, suggesting that cefovecin may have a shorter duration of action in these species than in cats and dogs. Cefovecin binding to plasma proteins in thawed, frozen equine plasma was also undertaken for assay quality control and the median (range) plasma protein binding (at 10 µg/mL) was 95.6% (94.9-96.6%). Cefovecin was also administered to six koalas at 8 mg/kg subcutaneously and serial blood samples were collected at 3, 6, 24, 48, 72, 96 h thereafter. Cefovecin plasma concentrations were not quantifiable in four koalas and in the other two, the mean plasma concentration at t = 3 h was 1.04 ± 0.01 µg/mL. CONCLUSION: Because of the limited pharmacokinetic data generated, no further pharmacokinetic analysis was performed; however, a single injected bolus of cefovecin is likely to have a short duration of action in koalas (hours, rather than days).

Hematologic and serum biochemical reference intervals for wild eastern quolls (Dasyurus viverrinus): Variation by age, sex, and season.

BACKGROUND: The eastern quoll (Dasyurus viverrinus) is an endangered carnivorous marsupial that has recently suffered significant population declines. Several small captive breeding populations have been established, with plans to translocate wild and captive individuals to areas of their former distribution. Accordingly, hematologic and serum biochemical reference intervals (RIs) established from wild eastern quoll populations are essential for monitoring the health and disease status of both captive and wild populations, and to evaluate the health of individuals before, during, and after translocation. OBJECTIVES: We aimed to establish hematologic and serum biochemical RIs for wild eastern quolls, and examine the effects of age, sex, and season. METHODS: We collected a total of 202 hematologic samples, 309 packed cell volume samples, and 335 serum biochemical samples from 168 individual quolls between May 2011 and November 2013. Species-level RIs were established, as well as RIs of groups separated by age (juvenile, adult) and sex (adult male, adult female) using nonparametric, robust, and parametric methods. Seasonal variation in age- and sex-specific reference values was also assessed. RESULTS: Strong age and seasonal variation were evident in many hematologic and serum biochemical analytes, with significant variation observed in serum biochemical analytes between the sexes. CONCLUSIONS: The observed age, sex, and seasonal variation reflect differences in the timing of growth and reproductive stressors, which interact with seasonal energetic demands. Our findings highlight the importance of using age-, sex-, and season-specific RIs for clinical evaluation of eastern quolls, as species-level RIs will inadvertently smooth and mask important seasonal fluctuations that reflect reproductive status at different times.

High blood lead concentrations in captive Tasmanian devils (Sarcophilus harrisii): a threat to the conservation of the species?

BACKGROUND: The Tasmanian devil (Sarcophilus harrisii) is the world's largest extant marsupial carnivore. Since the emergence of devil facial tumour disease in 1996, the species has undergone a severe population decline. The insurance population (IP) was established in 2006 to build a disease-free captive population to maintain 95% of the wild Tasmanian devil genetic diversity for 50 years. Captive and semi-wild Tasmanian devils are fed with possum and wallaby meat provided by local hunters, who use lead ammunition. Lead ingestion can cause acute toxicity, including ataxia, coma and death, or chronic subclinical deleterious effects including decreased fertility. METHODS: We determined blood lead concentrations in 26 captive and 133 wild Tasmanian devils from various sites across Tasmania. RESULTS: Captive Tasmanian devils showed significantly higher blood lead concentrations than their conspecifics in the wild. In captivity, older animals had higher blood lead concentrations than young animals, which suggested regular exposure, as lead can accumulate in a living organism in the blood, soft tissues and bones. After a response measure was implemented by removing the heads and wounds containing lead from the diet, blood concentrations significantly decreased in animals at one of the captive study sites, supporting the suspicion of food as the source of lead. CONCLUSION: This study highlights the need to ensure meat fed to captive carnivores is not contaminated by lead, especially in the context of a conservation program breeding individuals in captivity, as for Tasmanian devils.

ERBB3: A potential serum biomarker for early detection and therapeutic target for devil facial tumour 1 (DFT1).

Devil Facial Tumour 1 (DFT1) is one of two transmissible neoplasms of Tasmanian devils (Sarcophilus harrisii) predominantly affecting their facial regions. DFT1's cellular origin is that of Schwann cell lineage where lesions are evident macroscopically late in the disease. Conversely, the pre-clinical timeframe from cellular transmission to appearance of DFT1 remains uncertain demonstrating the importance of an effective pre-clinical biomarker. We show that ERBB3, a marker expressed normally by the developing neural crest and Schwann cells, is immunohistohemically expressed by DFT1, therefore the potential of ERBB3 as a biomarker was explored. Under the hypothesis that serum ERBB3 levels may increase as DFT1 invades local and distant tissues our pilot study determined serum ERBB3 levels in normal Tasmanian devils and Tasmanian devils with DFT1. Compared to the baseline serum ERBB3 levels in unaffected Tasmanian devils, Tasmanian devils with DFT1 showed significant elevation of serum ERBB3 levels. Interestingly Tasmanian devils with cutaneous lymphoma (CL) also showed elevation of serum ERBB3 levels when compared to the baseline serum levels of Tasmanian devils without DFT1. Thus, elevated serum ERBB3 levels in otherwise healthy looking devils could predict possible DFT1 or CL in captive or wild devil populations and would have implications on the management, welfare and survival of Tasmanian devils. ERBB3 is also a therapeutic target and therefore the potential exists to consider modes of administration that may eradicate DFT1 from the wild.

Leptin levels, seasonality and thermal acclimation in the Microbiotherid marsupial Dromiciops gliroides: Does photoperiod play a role?

Mammals of the Neotropics are characterized by a marked annual cycle of activity, which is accompanied by several physiological changes at the levels of the whole organism, organs and tissues. The physiological characterization of these cycles is important, as it gives insight on the mechanisms by which animals adjust adaptively to seasonality. Here we studied the seasonal changes in blood biochemical parameters in the relict South American marsupial Dromiciops gliroides ("monito del monte" or "little mountain monkey"), under semi-natural conditions. We manipulated thermal conditions in order to characterize the effects of temperature and season on a battery of biochemical parameters, body mass and adiposity. Our results indicate that monitos experience an annual cycle in body mass and adiposity (measured as leptin levels), reaching a maximum in winter and a minimum in summer. Blood biochemistry confirms that the nutritional condition of animals is reduced in summer instead of winter (as generally reported). This was coincident with a reduction of several biochemical parameters in summer, such as betahydroxybutyrate, cholesterol, total protein concentration and globulins. Monitos seem to initiate winter preparation during autumn and reach maximum body reserves in winter. Hibernation lasts until spring, at which time they use fat reserves and become reproductively active. Sexual maturation during summer would be the strongest energetic bottleneck, which explains the reductions in body mass and other parameters in this season. Overall, this study suggests that monitos anticipate the cold season by a complex interaction of photoperiodic and thermal cues.

Hematologic and serum biochemical changes associated with Devil Facial Tumor Disease in Tasmanian Devils.

BACKGROUND: Devil Facial Tumor Disease (DFTD) is an infectious tumor causing significant population declines in wild Tasmanian Devils. While clinical assessment and pathology have been well reported for DFTD, there is a lack of information on hematologic and biochemical alterations associated with DFTD. OBJECTIVES: The purpose of the study was to determine hematologic and serum biochemical variation in healthy, wounded, and DFTD-affected Tasmanian Devils. METHODS: Blood samples were collected from wild Tasmanian Devils at 5 sites in Tasmania. Hematology and clinical biochemistry variables were compared between clinically healthy, wounded, and DFTD-affected devils. Differences were also analyzed among stages of DFTD, including individuals pre- and postclinical signs developing, and between ulcerated and nonulcerated DFTD tumors. RESULTS: Statistically significantly increased counts in WBC, neutrophils, and platelets, and concentration of fibrinogen, as well as decreased counts in lymphocytes, erythrocytes, and HGB concentration were observed in DFTD-affected devils compared to healthy devils. Activities of ALP, ALT, and GLDH, concentrations of sodium, potassium and albumin, and sodium-to-potassium ratio and albumin-to-globulin ratio were significantly lower, and AST activity was significantly higher in animals with DFTD when compared to clinically healthy animals. No significant differences were found among stages of DFTD or ulcerated and nonulcerated tumors. CONCLUSIONS: The differences in hematology and clinical chemistry variables in devils with DFTD compared to healthy devils are nonspecific and reflective of acute phase response and inflammation, and anemia of chronic disease. Similar changes are observed with wounds but to a lesser extent.

HEMATOLOGICAL AND SERUM BIOCHEMICAL VALUES IN ANESTHETIZED CAPTIVE TASMANIAN DEVILS (SARCOPHILUS HARRISII).

The Tasmanian devil (Sarcophilus harrisii) population has decreased by estimates of 80% in the past 20 yr due to the effects of devil facial tumor disease (DFTD). In the process of creating a DFTD-free insurance population, the captive population and the number of institutions housing devils worldwide has increased tremendously. In order to provide the best husbandry and veterinary care for these captive animals, it is essential to know normal hematology and biochemistry values for the species. Baseline reference intervals (RIs) were determined for hematology and biochemistry variables for 170 healthy anesthetized captive Tasmanian devils and significant sex and age differences were determined. Higher relative neutrophil counts, hemoglobin, mean corpuscular hemoglobin (MCH), creatinine, creatine phosphokinase, and cholesterol were seen in males compared to females, whereas higher white cell counts (WBC) and lymphocyte counts (absolute and relative) were seen in females. Subadults have higher red blood cell counts, WBC, lymphocytes (absolute and relative), calcium and phosphorus, alkaline phosphatase, glutamate dehydrogenase, glucose, and albumin than adults; whereas, adults have higher relative neutrophils, relative eosinophils, mean corpuscular volume, MCH, platelets, total solids, total plasma proteins, globulins, and chloride than subadults. This study provides a comprehensive report of hematology and serum biochemistry RIs for healthy captive anesthetized Tasmanian devils and offers invaluable diagnostic information to care for the growing captive population of this endangered marsupial.

Immunoglubolin dynamics and cancer prevalence in Tasmanian devils (Sarcophilus harrisii).

Immunoglobulins such as IgG and IgM have been shown to induce anti-tumour cytotoxic activity. In the present study we therefore explore total serum IgG and IgM expression dynamics in 23 known-aged Tasmanian devils (Sarcophilus harrisii) of which 9 where affected by Devil Facial Tumour Disease (DFTD). DFTD is clonally transmissible cancer that has caused massive declines in devil numbers. Our analyses revealed that IgM and IgG expression levels as well as IgM/IgG ratios decreased with increasing devil age. Neither age, sex, IgM nor IgG expression levels affected devil DFTD status in our analyses. However, devils with increased IgM relative to IgG expression levels had significantly lower DFTD prevalence. Our results therefore suggest that IgM/IgG ratios may play an important role in determining devil susceptibility to DFTD. We consequently propose that our findings warrant further studies to elucidate the underpinning(s) of devil IgM/IgG ratios and DFTD status.

Hematologic and serum biochemical reference intervals for wild Tasmanian devils (Sarcophilus harrisii).

BACKGROUND: The Tasmanian devil (Sarcophilus harrisii) is a carnivorous marsupial threatened with extinction by a fatally infectious cancer known as devil facial tumor disease (DFTD). Conservation efforts including captive breeding and island translocations are underway to address this threat. OBJECTIVES: The objectives of this study were to determine hematologic and serum biochemical reference intervals (RI) to aid in health assessment of Tasmanian devils, and to examine seasonal, sex, reproductive status and age variations. METHODS: We collected jugular blood samples from individual wild Tasmanian devils at 2 different locations over a 2-year period to determine hematologic and serum biochemical RI by nonparametric methods using the central 0.95 fraction. RESULTS: A total of 307 blood samples were collected from 187 devils. Significant age differences were found for ALP, CK, cholesterol, calcium, phosphate, albumin, globulins, albumin: globulin ratio, and glucose. Significant differences between sexes were observed for AST, creatinine, and potassium. Significant seasonal or reproductive status variation in adult males or breeding females were observed for PCV, HGB, RBC, MCHC, MCH, MCV, neutrophils and lymphocytes, fibrinogen, total plasma protein, AST, ALP, ALT, GLDH, bilirubin, urea, calcium, chloride, total protein, albumin, A:G, and glucose. CONCLUSIONS: Many of the differences observed between subgroups can be explained by growth requirements, reproductive demands, and seasonal effects on activity. This study has determined comprehensive RI for the Tasmanian devil, which will be used to assess animals targeted for captive breeding and translocations, or affected by DFTD.

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.

Effects of overnight captivity on antioxidant capacity and clinical chemistry of wild southern hairy-nosed wombats (Lasiorhinus latifrons).

An animal's antioxidant capacity is measured by its ability to quench reactive oxygen species (ROS). During everyday metabolism, antioxidants and ROS are in equilibrium with one another. In times of stress, an animal produces more ROS and therefore uses its antioxidant capacity more readily in order to maintain this equilibrium. When the production of ROS exceeds the antioxidant capacity, an animal will experience extensive oxidative stress, which can ultimately affect that animal's health. During experimental study of wild animals, it is often necessary to capture them for a short period of time. In order to obtain a measurement of the effects of short-term captivity on oxidative capacity in wild animals, a population of southern hairy-nosed wombats (Lasiorhinus latifrons) in Swan Reach, South Australia (34.57 degrees S, 139.60 degrees E), was studied. To assess the variation in antioxidant capacity, two assays, the ferric reducing ability of plasma and the trolox equivalent antioxidant capacity, were performed. A third assay, thiobarbituric acid reactive substances, was used to measure the effects of ROS. Measurements of the specific antioxidants uric acid, ascorbic acid, retinol, alpha-tocopherol, and superoxide dismutase were also performed. The biochemical parameters albumin, total protein, cholinesterase, creatinine, and urea were measured as indicators for health. Results showed a significant reduction in antioxidant capacity during the overnight period of captivity.

Boldness towards novelty and translocation success in captive-raised, orphaned Tasmanian devils.

Translocation of endangered animals is common, but success is often variable and/or poor. Despite its intuitive appeal, little is known with regards to how individual differences amongst translocated animals influence their post-release survival, growth, and reproduction. We measured consistent pre-release responses to novelty in a familiar environment (boldness; repeatability=0.55) and cortisol response in a group of captive-reared Tasmanian devils, currently listed as "Endangered" by the IUCN. The devils were then released at either a hard- or soft-release site within their mothers' population of origin, and individual growth, movement, reproduction (females only), and survival across 2-8 months post-release was measured. Sex, release method, cohort, behavior, and cortisol response did not affect post-release growth, nor did these factors influence the home range size of orphan devils. Final linear distances moved from the release site were impacted heavily by the release cohort, but translocated devils' movement overall was not different from that in the same-age wild devils. All orphan females of reproductive age were subsequently captured with offspring. Overall survival rates in translocated devils were moderate (∼42%), and were not affected by devil sex, release method, cohort, release weight, or pre-release cortisol response. Devils that survived during the study period were, however, 3.5 times more bold than those that did not (effect size r=0.76). Our results suggest that conservation managers may need to provide developmental conditions in captivity that promote a wide range of behaviors across individuals slated for wild release.

Further investigation of the blood characteristics of Australian Quoll (Dasyurus spp.) species.

BACKGROUND: The Eastern and Spotted-Tailed Quolls are "near threatened" Australian dasyurid marsupials that have undergone significant reduction in their geographic ranges in the past 200 years. Captive breeding and research colonies now exist, allowing further efforts to more fully understand the physiology of these carnivorous species. OBJECTIVES: The aims of the study were to provide a more detailed study of blood chemistry and differential WBC counts for Eastern and Spotted-Tailed Quolls, and to determine the influence by 3 biologic factors. METHODS: Blood samples were taken from conscious, captive, healthy Eastern Quolls. A small number of samples from Spotted-Tailed Quolls were also available and were included in the study for comparison. Blood chemistry and differential WBC counts were compared to determine season-, age-, and sex-related differences. RESULTS: For many of the analytes, blood chemistry results were comparable to other marsupial ranges, and no significant differences between sexes were detected (P > .05). Seasonal differences were determined for total bilirubin, glucose, creatinine, and potassium concentrations in the Eastern Quoll. Generally, higher concentrations of these analytes were observed in the summer; however, amylase activity was significantly higher in autumn (southern hemisphere). Eastern Quolls one year of age and younger had significantly (P < .05) higher ALP activities than older animals. CONCLUSIONS: The normal ranges determined in this study can be used to assess clinical health of quolls and will assist with captive management and future reintroduction programs to the wild.

Effects and treatment of sarcoptic mange in southern hairy-nosed wombats (Lasiorhinus latifrons).

We examined the clinical and cellular effects of sarcoptic mange on southern hairy-nosed wombats (SHNW, Lasiorhinus latifrons) and the effectiveness of a single dose of ivermectin as a treatment for captive and wild animals. Wambats were caught at three sites in South Australia between April and August 2005 and blood and skin samples were collected. Hematology, biochemistry, and protein electrophoresis reference intervals were determined for healthy and diseased SHNW. Diseased SHNW had significantly higher white blood cell counts, neutrophils, lymphocytes, and total protein but lower red blood cell counts, hemoglobin, hematocrit, and creatinine. Microscopic investigation indicated substantial hyperplasia, hyperkeratosis, and fluid infiltration into the dermis and epidermis of diseased animals. Conclusions on the efficacy of a single dose of ivermectin were limited by low sample size (n=5, two captive and three wild SHNW) and are preliminary. However, ivermectin effectively treated mild, but not severe, mange in wild SHNW and severe mange in captive animals. This study has implications for the conservation and management of SHNW and the broader Vombatidae family.

Hematologic characteristics of the woylie (Bettongia penicillata ogilbyi).

An accurate assessment of animal health is fundamental to disease investigation in wildlife. Blood samples (n = 609) from several populations of the endangered woylie or brush-tailed bettong (Bettongia penicillata ogilbyi), collected between March 2006 and April 2010 in Western Australia and South Australia, were used to establish hematologic reference ranges. Differences between populations, sexes, and seasons were also investigated. Significant sex differences in hematocrit, red blood cell, total white blood cell, neutrophil, lymphocyte, and eosinophil counts were evident in at least one population. Generally, males had higher hematocrit and blood cell concentrations than did females. A positive association of the erythron parameters with rainfall was also detected. The hematologic characteristics of woylie populations described in this study greatly increase knowledge of the health status in these populations. The data also represent a baseline to enable monitoring and detection of changes in the health status in these populations as well as representing a valid dataset for comparison with hematologic investigations in other macropods and marsupials.

Hematologic and serum biochemical reference ranges and an assessment of exposure to infectious diseases prior to translocation of the threatened western ringtail possum (Pseudocheirus occidentalis).

Health screening of animals before translocation is important to minimize the risk of pathogen transmission between sites and species. Reintroduction has been incorporated into management of the endangered western ringtail possum (Pseudocheirus occidentalis) to mitigate for habitat loss within the species' core range in southwestern Australia. Between November 2005 and March 2008 we screened 47 wild and 24 captive P. occidentalis and 68 sympatric common brushtail possums (Trichosurus vulpecula hypoleucus) for infectious diseases that might compromise possum survival or fecundity at translocation sites. We found no evidence that infectious disease limits translocation success, and neither possum species showed evidence of infection with Salmonella spp., Toxoplasma gondii, Leptospira spp., or Chlamydophila spp. Antigen of Cryptococcus gattii was detected in one T. v. hypoleucus but was not of pathologic significance. Hematologic and serum biochemical reference ranges were determined for 81 wild and 24 captive P. occidentalis. Site differences were identified for red blood cell count, hemoglobin, albumin, urea, and globulin, suggesting that habitat quality or nutrient intake may vary among sites. Differences between wild and captive values were found for several parameters. These data are useful for health evaluations of injured P. occidentalis and the future monitoring of wild populations.

Development and evaluation of electroejaculation techniques in the Tasmanian devil (Sarcophilus harrisii).

Electroejaculation (EEJ) has been used successfully to collect samples suitable for genome resource banking from a variety of endangered wildlife species. Ejaculates can also be used to evaluate the reproductive potential of individuals and provide information on seminal characteristics to aid in the development of sperm cryopreservation techniques. Electroejaculation techniques used for marsupial and eutherian species were tested on Tasmanian devils (n=35). Spermic ejaculates were collected in 54% (19/35) of EEJ attempts. Spermic ejaculates were low in volume (3.9±6.5×10(2) µL, range 10-3000 µL) and contained low numbers of spermatozoa (3.3±7.8×10(3) spermatozoa per ejaculate, range 6-33000). The osmolality and pH of presumptive urine-free ejaculates were 389±130 mOsm kg(-1) (range 102-566) and 7.0±0.9 (range 6.0-8.0), respectively. Prostatic bodies were observed in 79% (26/33) of ejaculates. Episodic fluctuations in serum testosterone concentrations were not detected during the EEJ procedure (P>0.05). Increases observed in serum cortisol concentrations during EEJ were less (P<0.05) than those observed after an adrenalcorticotropic hormone challenge and diurnal variation suggested that cortisol concentrations are greater during the day than at night (P<0.05). This information can be used to provide range values for the future examination of basic endocrine responses and the adrenal-pituitary axis of this species. This study also demonstrated that spermatozoa-rich devil electroejaculates are more difficult to obtain and poorer in quality than those of other marsupials.