Wild Devils Research Articles

Lead exposure and source attribution for a mammalian scavenger before and after a culling program

Lead-based ammunition is a significant source of environmental lead and threatens species that scavenge lead-shot carcasses, particularly in areas with intensive shooting. With the impacts of lead on avian scavengers well established, there is increasing focus on the effects of lead on mammalian scavengers. We investigated lead exposure in a morphologically specialized mammalian scavenger, the Tasmanian devil (Sarcophilus harrisii), by analyzing their blood lead levels (BLLs) before and after a marsupial culling program using linear mixed effects models. We compared lead isotope signatures in devil blood to those in the culling ammunition to inform potential source attributions. We sampled 23 devils before culling and 15 after culling, finding no significant difference in mean BLLs pre and post-culling. However, devils captured closer to forestry coupes where culling had occurred had higher BLLs, and a greater proportion of devils displayed elevated BLLs post-culling (33 % compared to 18 % pre-culling). The highest BLL (7.93 μg/dL) was found in a devil post-culling and this individual had lead isotope signatures that matched the ammunition samples analyzed, suggesting the individual was exposed to lead from scavenging on culled carcasses. While 18 % of the devil blood lead samples had isotope signatures consistent with the ammunition samples, most were measurably different, indicating other sources of lead in the landscape. BLLs in our study landscape were similar to published BLLs for wild devils across Tasmania. That said, lead isotope signatures in the blood of individual devils sampled both before and after culling shifted closer to those of ammunition samples post-culling. Our results indicate that while some individual devils may have been exposed to lead from culling, most devils in the landscape did not show evidence of recent exposure. However, even low lead levels can adversely impact wildlife health and immunity, a particular concern for devils, a species endangered by disease.

Investigating scent communication and latrine use to inform translocation of the Tasmanian devil

Translocation programs for endangered mammalian carnivores face numerous challenges, including the potential for lethal interactions between releasees and resident conspecifics. Here we evaluate whether familiarizing residents with the scent of releasees might be an effective strategy for reducing aggression following Tasmanian devil (Sarcophilus harrisii) translocation. Tasmanian devils live in loose social networks in which residents have overlapping home ranges and use communal latrines. In a controlled ex situ experiment, we first found that devils readily distinguished between the feces of familiar and unfamiliar conspecifics. Next, we swapped feces between devils slated for release and wild resident devils, seeding a random half of communal latrines in the wild with the feces of releasee prior to their release, and used motion-activated video cameras to record devil visits. Wild resident devil visitation frequency and olfactory communication behaviors initially increased at experimental latrine sites where feces were deposited but not at control sites. After release, the translocated devils used both types of latrine sites and spent more time investigating experimental sites, but investigatory and scent-marking behaviors of the wild resident devils decreased, suggesting that they had become familiar with the releasees through scent. We further show that Tasmanian devil latrine sites are strongly associated with ecotones and the presence of shrubs. These findings contribute to our understanding of Tasmanian devil scent ecology and suggest that devil translocations conducted into existing populations would benefit from familiarization through latrine scent manipulation. More broadly, our results suggest an important role for scent ecology in carnivore translocation programs.Significance statementA conservation management action that is being trialed for endangered Tasmanian devils is to breed devils in human care and reinforce dwindling small resident populations through translocation. However, with limited social interactions, unfamiliar devils may interact aggressively, increasing injury and reducing translocation success. Scent communication is nearly ubiquitous in mammals and may be a prerequisite for management of social aggression. Almost nothing is known about devil scent ecology. Our discovery that devils become familiarized with conspecifics through feces, combined with findings that wild devils that are exposed to translocated devil scent in advance of release show reduced interest in scent-seeded latrines, even after new devils are released, suggests that devils familiarized through scent in advance of relocation may exhibit reduced aggression. These results provide an avenue for improving the success of translocations for devils and other at-risk solitary mammalian carnivores.

Faecal glucocorticoid metabolite levels of intensive captive, free‐range captive and wild Tasmanian devils

AbstractCaptivity can alter the stress physiology and behaviour of an animal in both the short‐ and long‐term through repetitive exposure to novel stressors and, subsequently, may reduce the success of conservation efforts such as translocation and reintroduction. The Tasmanian devil (Sarcophilus harrisii) is threatened with extinction from a fatal facial tumour disease which has led to the establishment of an insurance meta‐population designed for future reintroductions of disease‐free devils. The meta‐population is comprised of intensive captive and free‐range captive environments; however, no study has yet examined the long‐term physiological implications of captivity on devils. We used non‐invasive faecal glucocorticoid metabolite (FGM) monitoring to determine if there were any differences in adrenal activity among intensive captive, free‐range captive and wild devils. FGM levels were not age or sex dependent, and we found that all population types had similar intra‐population variability and mean FGMs. In conclusion, both types of captive environment appear to maintain stress profiles similar to wild devils.

Restoring faith in conservation action: Maintaining wild genetic diversity through the Tasmanian devil insurance program

SummaryConservation breeding programs aim to maintain 90% wild genetic diversity, but rarely assess functional diversity. Here, we compare both genome-wide and functional diversity (in over 500 genes) of Tasmanian devils (Sarcophilus harrisii) within the insurance metapopulation and across the species’ range (64,519 km2). Populations have declined by 80% since 1996 due to a contagious cancer, devil facial tumor disease (DFTD). However, predicted local extinctions have not occurred. Recent suggestions of selection for “resistance” alleles in the wild precipitated concerns that insurance population devils may be unsuitable for translocations. Using 830 wild samples collected at 31 locations between 2012 and 2021, and 553 insurance metapopulation devils, we show that the insurance metapopulation is representative of current wild genetic diversity. Allele frequencies at DFTD-associated loci were not substantially different between captive and wild devils. Methods presented here are valuable for others investigating evolutionary potential in threatened species, particularly ones under significant selective pressures.

Evaluation of oral baits and distribution methods for Tasmanian devils (Sarcophilus harrisii)†

Context Diseases are increasingly contributing to wildlife population declines. Tasmanian devil (Sarcophilus harrisii) populations have locally declined by 82%, largely owing to the morbidity and mortality associated with two independent transmissible devil facial tumours (DFT1 and DFT2). Toxic baits are often used as a management tool for controlling vertebrate pest populations in Australia, but in other areas of the world, oral baits are also used to deliver vaccines or pharmaceuticals to wildlife. Aim Our goal was to evaluate the potential use of edible baits as vehicles for vaccine delivery to Tasmanian devils. Method We first tested bait palatability with captive devils. Bait interactions were recorded, and consumption and bait interaction behaviours were quantified. We next trialled baits containing inert capsules as potential vaccine containers in captivity. After confirming bait palatability in captivity, ground baiting was trialled at six field sites and monitored using camera traps. Finally, an automated bait dispenser was trialled at field sites to attempt to limit bait consumption by non-target species. Key results Captive devils consumed all types of placebo baits, but consumed a higher percentage of ruminant- and fish-based baits than cereal-based baits. Captive devils also consumed inert capsules inserted into placebo baits. Ground-baiting trials in the field showed that 53% of baits were removed from bait stations, with 76% of the removals occurring on the first night. Devils were suspected or confirmed to remove about 7% of baits compared with 93% by non-target species. We also evaluated an automated bait dispenser, which reduced bait removal by non-target species and resulted in over 50% of the baits being removed by devils. Conclusions This study demonstrated that captive and wild devils will accept and consume placebo versions of commercial baits. Bait dispensers or modified baits or baiting strategies are needed to increase bait uptake by devils. Implications Bait dispensers can be used at a regional scale to deliver baits to devils. These could potentially be used as vaccine-delivery vehicles to mitigate the impacts of disease on devil populations.

Torn limb from limb: the ethology of prey-processing in Tasmanian devils (

The success of carnivorous mammals is determined not only by their ability to locate and kill prey, but also their efficiency at consuming it. Breaking large prey into small pieces is challenging due to the strong and tough materials that make up a carcass (e.g. hide, muscle, and bone). Carnivores therefore require a diverse suite of prey-processing behaviours to utilise their catch. Tasmanian devils are Australia’s only large marsupial scavengers and have the ability to consume almost all of a carcass. To determine how they do this we analysed 5.5 hours of footage from 21 captive and wild devils feeding at carcasses. We documented 6320 bouts of 12 distinct prey-processing behaviours, performed at frequencies that varied throughout feeds and between groups. The time point in the feed influenced the types of behaviours used. This is likely due to changing prey size, as different techniques appear better suited to handling whole carcasses or large pieces (pulling and pinning) or smaller pieces (holding and manipulating). Group size impacted the frequency of social pulling behaviours, which increased with the number of animals. Our findings highlight the range of prey-processing behaviours performed by scavenging devils when handling, breaking down, and consuming a carcass. The devils’ repertoire shares similarities with large carnivores that handle and consume whole carcasses as well as small carnivores that are adept in grasping and handling smaller prey.

Wearing the devil down: Rate of tooth wear varies between wild and captive Tasmanian devils.

Mammalian carnivores rely on their sharp teeth to effectively kill and consume prey. However, over time this causes wear and breakage that alters tooth shape, reducing their effectiveness. Extreme tooth wear and damage is especially prevalent in species that scavenge carcasses, like the Tasmanian devil (Sarcophilus harrisii), which are well known for their voracious appetites and ability to consume almost all of a carcass, including bone. In this study, we comprehensively describe tooth wear in captive and wild devils to look for differences in the patterns and rate of wear between these environments. To do this we surveyed tooth condition in skulls from 182 wild and 114 captive devils for which age was estimated using canine over-eruption. We found the types of tooth wear documented were the same in captive and wild devils, but captive animals have less severe wear than wild devils of the same estimated age. There was no difference in the proportion of captive or wild individuals with broken canine or molar teeth; however, breakage occurred at a younger age in wild devils. Although not considered anomalous or harmful, this indicates a difference in the way teeth are being used and/or the foods consumed between captive and wild devils. We hypothesize how these results relate to differences in diet or behavior that may stem from their various feeding environments, for example, higher quality food (fresh, whole, and yet to be scavenged carcasses) provided to captive devils likely causes less wear. Further, we support management options that closely replicate wild diet items and behaviors suitable for a long-term insurance population.

Infectious disease and sickness behaviour: tumour progression affects interaction patterns and social network structure in wild Tasmanian devils.

Infectious diseases, including transmissible cancers, can have a broad range of impacts on host behaviour, particularly in the latter stages of disease progression. However, the difficulty of early diagnoses makes the study of behavioural influences of disease in wild animals a challenging task. Tasmanian devils (Sarcophilus harrisii) are affected by a transmissible cancer, devil facial tumour disease (DFTD), in which tumours are externally visible as they progress. Using telemetry and mark-recapture datasets, we quantify the impacts of cancer progression on the behaviour of wild devils by assessing how interaction patterns within the social network of a population change with increasing tumour load. The progression of DFTD negatively influences devils' likelihood of interaction within their network. Infected devils were more active within their network late in the mating season, a pattern with repercussions for DFTD transmission. Our study provides a rare opportunity to quantify and understand the behavioural feedbacks of disease in wildlife and how they may affect transmission and population dynamics in general.

Inbreeding depression in one of the last DFTD-free wild populations of Tasmanian devils.

BackgroundVulnerable species experiencing inbreeding depression are prone to localised extinctions because of their reduced fitness. For Tasmanian devils, the rapid spread of devil facial tumour disease (DFTD) has led to population declines and fragmentation across the species’ range. Here we show that one of the few remaining DFTD-free populations of Tasmanian devils is experiencing inbreeding depression. Moreover, this population has experienced a significant reduction in reproductive success over recent years.MethodsWe used 32 microsatellite loci to examine changes in genetic diversity and inbreeding in the wild population at Woolnorth, alongside field data on breeding success from females to test for inbreeding depression.ResultsWefound that maternal internal relatedness has a negative impact on litter sizes. The results of this study imply that this population may be entering an extinction vortex and that to protect the population genetic rescue should be considered. This study provides conservation managers with useful information for managing wild devils and provides support for the “Wild Devil Recovery Program”, which is currently augmenting small, isolated populations.

Salmonella in captive and wild Tasmanian devils (Sarcophilus harrisii) in Tasmania.

Translocation of Tasmanian devils (Sarcophilus harrisii) is a common strategy for recovery of the species as carried out by the Save the Tasmanian Devil Program. Dasyurids including the endangered Tasmanian devil are well known to asymptomatically harbour the zoonotic bacteria Salmonella enterica in their intestinal tracts. Testing for Salmonella is a routine component of pretranslocation health testing, so a statewide microbiological survey of captive and wild devils was implemented in order to understand prevalence and common Salmonella serotypes, and inform decision-making when positive cultures are identified. This preliminary study identified a significantly higher proportion of Salmonella isolations in wild compared with captive devils. Mississippi and Typhimurium were the most common serotypes, followed by Lexington, Bovismorbificans, Kottbus and Amsterdam. Given the common finding of Salmonella in wild devils and the range of serotypes involved, in addition to numerous isolations in domestic species and humans, it is unlikely that the release of small numbers of captive devils to the wild in Tasmania poses a significant risk to the destination ecosystem. Ongoing monitoring of devils is required as the stress of acclimatisation could predispose devils to clinical disease. Appropriate personal protective attire is pertinent to protect personnel handling animals from this zoonotic infection.

Looking like the locals - gut microbiome changes post-release in an endangered species

BackgroundCaptivity presents extreme lifestyle changes relative to the wild, and evidence of microbiome dysbiosis in captive animals is growing. The gut microbiome plays a crucial role in host health. Whilst captive breeding and subsequent reintroduction to the wild is important for conservation, such efforts often have limited success. Post-release monitoring is essential for assessing translocation success, but changes to the microbiome of released individuals are poorly understood. The Tasmanian devil was previously shown to exhibit loss of microbiome diversity as a result of intense captive management. This current study examines changes in the devil gut microbiome in response to translocation and aims to determine if perturbations from captivity are permanent or reversible.MethodsUsing 16S rRNA amplicon sequencing, we conducted temporal monitoring of the gut microbiome of released devils during two translocation events, captive-to-wild and wild-to-wild. To investigate whether the microbiome of the released devils changed following translocation, we characterized their microbiome at multiple time points during the translocation process over the course of 6–12 months and compared them to the microbiome of wild incumbent devils (resident wild-born devils at the respective release sites).ResultsWe showed that the pre-release microbiome was significantly different to the microbiome of wild incumbent animals, but that the microbiomes of animals post-release (as early as 3 to 4 weeks post-release) were similar to wild incumbents. The gut microbiome of released animals showed significant compositional shifts toward the wild incumbent microbiome of both translocation events.ConclusionOur results suggest that the devil gut microbiome is dynamic and that loss of microbiome diversity in captivity can be restored following release to the wild. We recommend the broader application of microbiome monitoring in wildlife translocation programs to assess the impacts of translocation on animal microbiomes.

MHC-associated mate choice under competitive conditions in captive versus wild Tasmanian devils

AbstractMate choice contributes to driving evolutionary processes when animals choose breeding partners that confer genetic advantages to offspring, such as increased immunocompetence. The major histocompatibility complex (MHC) is an important group of immunological molecules, as MHC antigens bind and present foreign peptides to T-cells. Recent studies suggest that mates may be selected based on their MHC profile, leading to an association between an individual’s MHC diversity and their breeding success. In conservation, it may be important to consider mate choice in captive breeding programs, as this mechanism may improve reproductive rates. We investigated the reproductive success of Tasmanian devils in a group housing facility to determine whether increased MHC-based heterozygosity led individuals to secure more mating partners and produce more offspring. We also compared the breeding success of captive females to a wild devil population. MHC diversity was quantified using 12 MHC-linked microsatellite markers, including 11 previously characterized markers and one newly identified marker. Our analyses revealed that there was no relationship between MHC-linked heterozygosity and reproductive success either in captivity or the wild. The results of this study suggest that, for Tasmanian devils, MHC-based heterozygosity does not produce greater breeding success and that no specific changes to current captive management strategies are required with respect to preserving MHC diversity.

Fecal Viral Diversity of Captive and Wild Tasmanian Devils Characterized Using Virion-Enriched Metagenomics and Metatranscriptomics.

The Tasmanian devil is an endangered carnivorous marsupial threatened by devil facial tumor disease (DFTD). While research on DFTD has been extensive, little is known about viruses in devils and whether any are of potential conservation relevance for this endangered species. Using both metagenomics based on virion enrichment and sequence-independent amplification (virion-enriched metagenomics) and metatranscriptomics based on bulk RNA sequencing, we characterized and compared the fecal viromes of captive and wild devils. A total of 54 fecal samples collected from two captive and four wild populations were processed for virome characterization using both approaches. In total, 24 novel marsupial-related viruses, comprising a sapelovirus, astroviruses, rotaviruses, picobirnaviruses, parvoviruses, papillomaviruses, polyomaviruses, and a gammaherpesvirus, were identified, as well as known mammalian pathogens such as rabbit hemorrhagic disease virus 2. Captive devils showed significantly lower viral diversity than wild devils. Comparison of the two virus discovery approaches revealed substantial differences in the number and types of viruses detected, with metatranscriptomics better suited for RNA viruses and virion-enriched metagenomics largely identifying more DNA viruses. Thus, the viral communities revealed by virion-enriched metagenomics and metatranscriptomics were not interchangeable and neither approach was able to detect all viruses present. An integrated approach using both virion-enriched metagenomics and metatranscriptomics constitutes a powerful tool for obtaining a complete overview of both the taxonomic and functional profiles of viral communities within a sample.IMPORTANCE The Tasmanian devil is an iconic Australian marsupial that has suffered an 80% population decline due to a contagious cancer, devil facial tumor disease, along with other threats. Until now, viral discovery in this species has been confined to one gammaherpesvirus (dasyurid herpesvirus 2 [DaHV-2]), for which captivity was identified as a significant risk factor. Our discovery of 24 novel marsupial-associated RNA and DNA viruses, and that viral diversity is lower in captive than in wild devils, has greatly expanded our knowledge of gut-associated viruses in devils and provides important baseline information that will contribute to the conservation and captive management of this endangered species. Our results also revealed that a combination of virion-enriched metagenomics and metatranscriptomics may be a more comprehensive approach for virome characterization than either method alone. Our results thus provide a springboard for continuous improvements in the way we study complex viral communities.

Conserving adaptive potential: lessons from Tasmanian devils and their transmissible cancer.

Maintenance of adaptive genetic variation has long been a goal of management of natural populations, but only recently have genomic tools allowed identification of specific loci associated with fitness-related traits in species of conservation concern. This raises the possibility of managing for genetic variation directly relevant to specific threats, such as those due to climate change or emerging infectious disease. Tasmanian devils (Sarcophilus harrisii) face the threat of a transmissible cancer, devil facial tumor disease (DFTD), that has decimated wild populations and led to intensive management efforts. Recent discoveries from genomic and modeling studies reveal how natural devil populations are responding to DFTD, and can inform management of both captive and wild devil populations. Notably, recent studies have documented genetic variation for disease-related traits and rapid evolution in response to DFTD, as well as potential mechanisms for disease resistance such as immune response and tumor regression in wild devils. Recent models predict dynamic persistence of devils with or without DFTD under a variety of modeling scenarios, although at much lower population densities than before DFTD emerged, contrary to previous predictions of extinction. As a result, current management that focuses on captive breeding and release for maintaining genome-wide genetic diversity or demographic supplementation of populations could have negative consequences. Translocations of captive devils into wild populations evolving with DFTD can cause outbreeding depression and/or increases in the force of infection and thereby the severity of the epidemic, and we argue that these risks outweigh any benefits of demographic supplementation in wild populations. We also argue that genetic variation at loci associated with DFTD should be monitored in both captive and wild populations, and that as our understanding of DFTD-related genetic variation improves, considering genetic management approaches to target this variation is warranted in developing conservation strategies for Tasmanian devils.

Inducible IFN-γ Expression for MHC-I Upregulation in Devil Facial Tumor Cells.

The Tasmanian devil facial tumor (DFT) disease has led to an 80% reduction in the wild Tasmanian devil (Sarcophilus harrisii) population since 1996. The limited genetic diversity of wild devils and the lack of MHC-I expression on DFT cells have been implicated in the lack of immunity against the original DFT clonal cell line (DFT1). Recently, a second transmissible tumor of independent origin (DFT2) was discovered. Surprisingly, DFT2 cells do express MHC-I, but DFT2 cells appear to be on a trajectory for reduced MHC-I expression in vivo. Thus, much of the ongoing vaccine-development efforts and conservation plans have focused on MHC-I. A major limitation in conservation efforts is the lack of species-specific tools to understand Tasmanian devil gene function and immunology. To help fill this gap, we developed an all-in-one Tet-Off vector system to regulate expression of IFN-γ in DFT cells (DFT1.Tet/IFN-γ). IFN-γ can have negative effects on cell proliferation and viability; thus, doxycycline was used to suppress IFN-γ production whilst DFT1.Tet/IFN-γ cells were expanded in cell culture. Induction of IFN-γ following removal of doxycycline led to upregulation of MHC-I but also the inhibitory checkpoint molecule PD-L1. Additionally, DFT1.Tet/IFN-γ cells were capable of stimulating MHC-I upregulation on bystander wild type DFT cells in co-culture assays in vitro. This system represents a major step forward in DFT disease immunotherapy and vaccine development efforts, and ability to understand gene function in devils. Importantly, the techniques are readily transferable for testing gene function in DFT2 cells and other non-traditional species.

Two Decades of the Impact of Tasmanian Devil Facial Tumor Disease.

The Tasmanian devil, a marsupial carnivore, has been restricted to the island state of Tasmania since its extinction on the Australian mainland about 3000years ago. In the past two decades, this species has experienced severe population decline due to the emergence of devil facial tumor disease (DFTD), a transmissible cancer. During these 20years, scientists have puzzled over the immunological and evolutionary responses by the Tasmanian devil to this transmissible cancer. Targeted strategies in population management and disease control have been developed as well as comparative processes to identify variation in tumor and host genetics. A multi-disciplinary approach with multi-institutional teams has produced considerable advances over the last decade. This has led to a greater understanding of the molecular pathogenesis and genomic classification of this cancer. New and promising developments in the Tasmanian devil's story include evidence that most immunized, and some wild devils, can produce an immune response to DFTD. Furthermore, epidemiology combined with genomic studies suggest a rapid evolution to the disease and that DFTD will become an endemic disease. Since 1998 there have been more than 350 publications, distributed over 37 Web of Science categories. A unique endemic island species has become an international curiosity that is in the spotlight of integrative and comparative biology research.

Molecular characterization of Cryptosporidium and Giardia from the Tasmanian devil (Sarcophilus harrisii).

The Tasmanian devil (Sarcophilus harrisii) is a carnivorous marsupial found only in the wild in Tasmania, Australia. Tasmanian devils are classified as endangered and are currently threatened by devil facial tumour disease, a lethal transmissible cancer that has decimated the wild population in Tasmania. To prevent extinction of Tasmanian devils, conservation management was implemented in 2003 under the Save the Tasmanian Devil Program. This study aimed to assess if conservation management was altering the interactions between Tasmanian devils and their parasites. Molecular tools were used to investigate the prevalence and diversity of two protozoan parasites, Cryptosporidium and Giardia, in Tasmanian devils. A comparison of parasite prevalence between wild and captive Tasmanian devils showed that both Cryptosporidium and Giardia were significantly more prevalent in wild devils (p < 0.05); Cryptosporidium was identified in 37.9% of wild devils but only 10.7% of captive devils, while Giardia was identified in 24.1% of wild devils but only 0.82% of captive devils. Molecular analysis identified the presence of novel genotypes of both Cryptosporidium and Giardia. The novel Cryptosporidium genotype was 98.1% similar at the 18S rDNA to Cryptosporidium varanii (syn. C. saurophilum) with additional samples identified as C. fayeri, C. muris, and C. galli. Two novel Giardia genotypes, TD genotype 1 and TD genotype 2, were similar to G. duodenalis from dogs (94.4%) and a Giardia assemblage A isolate from humans (86.9%). Giardia duodenalis BIV, a zoonotic genotype of Giardia, was also identified in a single captive Tasmanian devil. These findings suggest that conservation management may be altering host-parasite interactions in the Tasmanian devil, and the presence of G. duodenalis BIV in a captive devil points to possible human-devil parasite transmission.

Seasonality and breeding success of captive and wild Tasmanian devils (Sarcophilus harrisii)

The synchrony and timing of reproductive events are crucially important factors to maximize individual and offspring survival, especially in seasonal environments. To increase our understanding of the physiological basis of seasonality and the influence of associated environmental factors (maximum temperature, day length and rate of day length change associated with different latitudes) on reproduction in Tasmanian devils, we reviewed records and research data from captive facilities throughout Australia in comparison to those from a wild population study (1974–1987). Overall, breeding activity began 2 weeks earlier in the captive than the wild population (week 5.7 ± 0.6 versus week 7.7 ± 0.5 for devils entering into estrus during the first two week phase; n = 24 and n = 23 respectively). If the timing of reproductive activity is considered against absolute day length rather than date, both the captive and wild populations displayed similar distributions (12.9 ± 0.7 h versus 13.0 ± 0.7 h respectively; P < 0.01) confirming day length as a proximal cue involved in eliciting a physiological response to trigger seasonal reproductive activity regardless of location. Wild devils had a higher breeding success (75%; n = 169 versus 43%; n = 115) and larger litter size (3.4 ± 0.9 versus 2.8 ± 1.1 joeys per litter) than captive devils (P < 0.05). Mean maximum temperature at the onset of reproductive activity (P < 0.05) was higher for the captive than the wild population (28.1 ± 4.0 °C versus 22.3 ± 2.7 °C respectively). The drivers for reproductive success in captive Tasmanian devils are likely multifactorial, but our results suggest that elevated temperatures associated with shifts in breeding activity and geographical location should be examined further.

PD-L1 Is Not Constitutively Expressed on Tasmanian Devil Facial Tumor Cells but Is Strongly Upregulated in Response to IFN-γ and Can Be Expressed in the Tumor Microenvironment.

The devil facial tumor disease (DFTD) is caused by clonal transmissible cancers that have led to a catastrophic decline in the wild Tasmanian devil (Sarcophilus harrisii) population. The first transmissible tumor, now termed devil facial tumor 1 (DFT1), was first discovered in 1996 and has been continually transmitted to new hosts for at least 20 years. In 2015, a second transmissible cancer [devil facial tumor 2 (DFT2)] was discovered in wild devils, and the DFT2 is genetically distinct and independent from the DFT1. Despite the estimated 136,559 base pair substitutions and 14,647 insertions/deletions in the DFT1 genome as compared to two normal devil reference genomes, the allograft tumors are not rejected by the host immune system. Additionally, genome sequencing of two sub-strains of DFT1 detected greater than 15,000 single-base substitutions that were found in only one of the DFT1 sub-strains, demonstrating the transmissible tumors are evolving and that generation of neoantigens is likely ongoing. Recent evidence in human clinical trials suggests that blocking PD-1:PD-L1 interactions promotes antitumor immune responses and is most effective in cancers with a high number of mutations. We hypothesized that DFTD cells could exploit the PD-1:PD-L1 inhibitory pathway to evade antitumor immune responses. We developed recombinant proteins and monoclonal antibodies (mAbs) to provide the first demonstration that PD-1 binds to both PD-L1 and PD-L2 in a non-placental mammal and show that PD-L1 is upregulated in DFTD cells in response to IFN-γ. Immunohistochemistry showed that PD-L1 is rarely expressed in primary tumor masses, but low numbers of PD-L1+ non-tumor cells were detected in the microenvironment of several metastatic tumors. Importantly, in vitro testing suggests that PD-1 binding to PD-L1 and PD-L2 can be blocked by mAbs, which could be critical to understanding how the DFT allografts evade the immune system.

A review of parasites in the Tasmanian devil (Sarcophilus harrisii)

Threatened by devil facial tumour disease, the Tasmanian devil (Sarcophilus harrisii), a carnivorous marsupial confined to Tasmania, Australia, is the subject of conservation management under the Save the Tasmanian Devil Program. Conservation actions such as captive breeding and translocation may impact upon parasite ecology, presenting risk of increased disease through stress and impaired immunity, and by exposing hosts to parasites to which they are immunologically naive. Given the importance of parasites to ecosystem function, it has been argued from a biodiversity perspective that parasites should be conserved in their own right. In this review we describe current knowledge, and limitations in our knowledge, of Tasmanian devil parasites. We then discuss the potential for changes in host–parasite interactions as a result of host-population decline and conservation management, both generally and with examples from the Tasmanian devil. The review closes with a recommendation for a systematic evaluation of parasites in captive and wild devils to aid conservation of this host–parasite system in its entirety.