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 statement A 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.

ABSTRACTDisease is increasingly becoming a driver of wildlife population declines and extinction risk. Vaccines have been one of the most successful health interventions in human history, but few have been tested for mitigating wildlife disease. The transmissible cancer, devil facial tumour disease (DFTD), triggered the Tasmanian devil’s (Sarcophilus harrisii) inclusion on the international endangered species list. Development of a protective DFTD vaccine would provide a valuable management approach for conservation of the species. In 2016, 33 devils from a DFTD-free insurance population were given an experimental DFTD vaccination prior to their release on the north coast of Tasmania. The release site was already home to an incumbent population of devils, including some individuals with DFTD. To determine the efficacy of the vaccination protocol and the longevity of the response it induced, six trapping trips took place at the site over the 2.5 years following release. Eight of the 33 vaccinated devils were re-trapped, and six of those developed DFTD within the monitoring period. Despite the apparent lack of protection provided by the vaccine for the re-trapped devils, we observed several signs of immune activation not usually found in unvaccinated devils. Firstly, sera collected from the eight devils showed that anti-DFTD antibodies persisted for up to two years post vaccination. Secondly, tumour infiltrating lymphocytes were found in three out of four biopsies collected from vaccinated devils which contrasts with the “immune deserts” typical of DFT’s; only one out of twenty incumbent devils with DFTD trapped during the same period had a tumour biopsy exhibiting immune cell infiltrate. Thirdly, immunohistochemical analysis of tumour biopsies from the vaccinated devils identified the functional immune molecules associated with antigen presenting cells (MHC-II) and T cells (CD3), and the immune checkpoint molecule PD-1, all associated with anti-tumour immunity in other species. These results correlate with our previous study on captive devils in which a prophylactic vaccine primed the devil immune system and, following DFTD challenge and tumour growth, immunotherapy induced complete tumour regressions. The field trial results presented here provide further evidence that the devil immune system can be primed to recognise DFTD cells, but additional immune manipulation could be needed for complete protection or induction of tumour regressions.

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.

Background.Vulnerable 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.Methods.We 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.Results.We found 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.

Introduction: The Tasmanian devil (Sarcophilus harrisii) is the largest extant carnivorous marsupial. Since 1996, its population has declined by 77% primarily due to a clonal transmissible tumor, known as devil facial tumor (DFT1) disease. In 2014, a second transmissible devil facial tumor (DFT2) was discovered. DFT1 and DFT2 are nearly 100% fatal.Areas covered: We review DFT control approaches and propose a rabies-style oral bait vaccine (OBV) platform for DFTs. This approach has an extensive safety record and was a primary tool in large-scale rabies virus elimination from wild carnivores across diverse landscapes. Like rabies virus, DFTs are transmitted by oral contact, so immunizing the oral cavity and stimulating resident memory cells could be advantageous. Additionally, exposing infected devils that already have tumors to OBVs could serve as an oncolytic virus immunotherapy. The primary challenges may be identifying appropriate DFT-specific antigens and optimization of field delivery methods.Expert opinion: DFT2 is currently found on a peninsula in southern Tasmania, so an OBV that could eliminate DFT2 should be the priority for this vaccine approach. Translation of an OBV approach to control DFTs will be challenging, but the approach is feasible for combatting ongoing and future disease threats.

Globally, conservation translocations are common often using island, or 'virtual island' populations as a source site. We investigated demographic and genetic consequences of using a single source island population for multiple translocations. Our harvest selection protocol preserves the source population whilst providing individuals that are representative of the source population. Our study species, Tasmanian devils, are endangered due to an infectious cancer with an 80 % reduction in population size across their range. In 2012, an island population was established to protect the species. The island population was harvested three times with devils translocated to wild location across Tasmania (2016, N = 16; 2017, N = 33; 2018, N = 30). Efforts to harvest individuals demographically representative of the source population was achievable in 2017 and 2018, with a large male bias occurring in 2016 due to logistic constraints. Using internal relatedness (IR; proxy for heterozygosity), we showed that harvesting animals in equal-sized groups of high, medium and low IR, succeeded in maintaining genetic profiles of the source population over repeated harvests. There was little within-year variation in observed heterozygosity pre- and post-harvest compared to the harvest group, and only a small reduction (0.001) in heterozygosity across years. We used simulations to show that using a random selection would likely have eroded diversity relative to our structured approach (86.6% of iterations showed greater loss). Our method maintained a viable source population over multiple harvests, providing conservation managers with a tool for conserving single source populations used in threatened species translocations.

Devil facial tumour 1 (DFT1) is a transmissible cancer clone endangering the Tasmanian devil. The expansion of DFT1 across Tasmania has been documented, but little is known of its evolutionary history. We analysed genomes of 648 DFT1 tumours collected throughout the disease range between 2003 and 2018. DFT1 diverged early into five clades, three spreading widely and two failing to persist. One clade has replaced others at several sites, and rates of DFT1 coinfection are high. DFT1 gradually accumulates copy number variants (CNVs), and its telomere lengths are short but constant. Recurrent CNVs reveal genes under positive selection, sites of genome instability, and repeated loss of a small derived chromosome. Cultured DFT1 cell lines have increased CNV frequency and undergo highly reproducible convergent evolution. Overall, DFT1 is a remarkably stable lineage whose genome illustrates how cancer cells adapt to diverse environments and persist in a parasitic niche.

Background.Vulnerable 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.Methods.We 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.Results.We found 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.

Background. Vulnerable 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. Methods. We 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. Results. We found 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.

The Tasmanian devil is threatened by Devil Facial Tumour Disease (DFTD), a transmissible form of cancer that has reduced the population by over 80%. Persecution, extreme climate events, vehicle collision and habitat destruction also put pressure on this endangered species. The recovery effort to save the Tasmanian devil commenced over 15 years ago as a collaborative initiative between the Tasmanian government, the Australian government, the Zoo and Aquarium Association Australasia, and many research institutions. Saving the Tasmanian Devil documents the journey taken by partner organisations in discovering what DFTD is, the effect it has on wild devil populations, and the outcomes achieved through research and management actions. Chapters describe all aspects of devil conservation, including the captive devil populations, applied pathology, immunology and genetic research findings, adaptive management, and the importance of advocacy and partnerships. This book will provide management practitioners and conservation scientists with insight into the complexities of undertaking a program of this scale, and will also be of value to researchers, students and others interested in conservation.

Emerging infectious diseases are rising globally and understanding host-pathogen interactions during the initial stages of disease emergence is essential for assessing potential evolutionary dynamics and designing novel management strategies. Tasmanian devils (Sarcophilus harrisii) are endangered due to a transmissible cancer-devil facial tumour disease (DFTD)-that since its emergence in the 1990s, has affected most populations throughout Tasmania. Recent studies suggest that devils are adapting to the DFTD epidemic and that disease-induced extinction is unlikely. However, in 2014, a second and independently evolved transmissible cancer-devil facial tumour 2 (DFT2)-was discovered at the d'Entrecasteaux peninsula, in south-east Tasmania, suggesting that the species is prone to transmissible cancers. To date, there is little information about the distribution, epidemiology and effects of DFT2 and its interaction with DFTD. Here, we use data from monitoring surveys and roadkills found within and adjacent to the d'Entrecasteaux peninsula to determine the distribution of both cancers and to compare their epidemiological patterns. Since 2012, a total of 51 DFTD tumours have been confirmed among 26 individuals inside the peninsula and its surroundings, while 40 DFT2 tumours have been confirmed among 23 individuals, and two individuals co-infected with both tumours. All devils with DFT2 were found within the d'Entrecasteaux peninsula, suggesting that this new transmissible cancer is geographically confined to this area. We found significant differences in tumour bodily location in DFTD and DFT2, with non-facial tumours more commonly found in DFT2. There was a significant sex bias in DFT2, with most cases reported in males, suggesting that since DFT2 originated from a male host, females might be less susceptible to this cancer. We discuss the implications of our results for understanding the epidemiological and evolutionary interactions of these two contemporary transmissible cancers and evaluating the effectiveness of potential management strategies.

BACKGROUND:Devil facial tumour disease (DFTD) is a contagious cancer causing marked population declines in wild Tasmanian devils. In response to this threat, a captive insurance population has been established. This study investigated causes of death in captive Tasmanian devils.METHODS:Clinical and laboratory records of captive Tasmanian devils held in seven Tasmanian captive facilities were analysed for cause of death or severe morbidity requiring euthanasia.RESULTS:Neoplasia was found to be the most common cause of mortality/severe morbidity, accounting for 27/63 of deaths. Cutaneous lymphoma was the most frequently observed tumour (10/27), at a higher incidence than previously reported. The most common cause of severe morbidity, following neoplasia, was leucoencephalomyelopathy, which caused severe, progressive hindlimb paresis and ataxia.CONCLUSION:Neoplasia, specifically cutaneous lymphoma, and degenerative neurological conditions are the most frequent causes of death in captive Tasmanian devils in Tasmania. Further work to determine the aetiologies of these conditions, as well as effective treatments, would be valuable.

The New Holland mouse (Pseudomys novaehollandiae) was recognised to be at threat of extinction at an International, National, and Tasmanian State level in 2008, 2010, and 1995, respectively. The species inhabits the floristically diverse dry heathlands and associated vegetation communities that typically arise from mosaic burns. In this regard the New Holland mouse could be considered a flagship species for healthy heathlands. We present 18 years of search and capture records collected since 1998 from Tasmania to show that there has been a contraction over the last nine years from 16 to 2 in the number of sites from which the species has been detected, and more than a 99% reduction in capture-per-unit-effort. These results raise concern as to the viability of the species in Tasmania. Strength of inference regarding the magnitude and causes of decline in distribution and abundance would be improved by regular and systematic monitoring. The considerable overlap between the potential causes of the New Holland mouse's decline in Tasmania and regional declines in mammal assemblages nationally, suggest that recovery efforts for the New Holland mouse could have positive outcomes for all species inhabiting lowland dry heathlands in Australia.

In order to project and plan for the future of threatened species, conservationists require good quality estimates of basic population parameters, such as population sex ratios and breeding rates. These data can be difficult to obtain in many threatened species where pervasive threats perturb population dynamics. For Tasmanian devils, previous studies at several sites across Tasmania revealed demographic conse-quences of the fatal and transmissible devil facial tumour disease (DFTD). In the current analysis, we take advantage of broad-scale survey data across the state, since 2003, to examine the differences between DFTD-present and DFTD-absent populations at the landscape level. Consistent with expectations based on previous studies, we found that devils trapped in DFTD-present sites were significantly younger (based on tooth-wear age estimates) than those in DFTD-absent sites. Interestingly, we also found that females in DFTD-present sites were more likely to show evidence of breeding than females in DFTD-absent sites. This difference could not be attributed to differences in age or body condition. Devil populations are declining, so increased female breeding rate is unlikely to be successfully compensating for decreased lifespan as a result of DFTD at the population level. These data can be used to inform stochastic and evolu-tionary models of population dynamics for devils, and inform the assessment of conservation strategies for the species in the presence of disease.

Devil facial tumor disease (DFTD) is renowned for its successful evasion of the host immune system. Down regulation of the major histocompatabilty complex class I molecule (MHC-I) on the DFTD cells is a primary mechanism of immune escape. Immunization trials on captive Tasmanian devils have previously demonstrated that an immune response against DFTD can be induced, and that immune-mediated tumor regression can occur. However, these trials were limited by their small sample sizes. Here, we describe the results of two DFTD immunization trials on cohorts of devils prior to their wild release as part of the Tasmanian Government's Wild Devil Recovery project. 95% of the devils developed anti-DFTD antibody responses. Given the relatively large sample sizes of the trials (N = 19 and N = 33), these responses are likely to reflect those of the general devil population. DFTD cells manipulated to express MHC-I were used as the antigenic basis of the immunizations in both trials. Although the adjuvant composition and number of immunizations differed between trials, similar anti-DFTD antibody levels were obtained. The first trial comprised DFTD cells and the adjuvant combination of ISCOMATRIX™, polyIC, and CpG with up to four immunizations given at monthly intervals. This compared to the second trial whereby two immunizations comprising DFTD cells and the adjuvant combination ISCOMATRIX™, polyICLC (Hiltonol®) and imiquimod were given a month apart, providing a shorter and, therefore, more practical protocol. Both trials incorporated a booster immunization given up to 5 months after the primary course. A key finding was that devils in the second trial responded more quickly and maintained their antibody levels for longer compared to devils in the first trial. The different adjuvant combination incorporating the RNAase resistant polyICLC and imiquimod used in the second trial is likely to be responsible. The seroconversion in the majority of devils in these anti-DFTD immunization trials was remarkable, especially as DFTD is hallmarked by its immune evasion mechanisms. Microsatellite analyzes of MHC revealed that some MHC-I microsatellites correlated to stronger immune responses. These trials signify the first step in the long-term objective of releasing devils with immunity to DFTD into the wild.

Tasmanian devils have spawned two transmissible cancer clones, known as devil facial tumour 1 (DFT1) and devil facial tumour 2 (DFT2). DFT1 and DFT2 are transmitted between animals by the transfer of allogeneic contagious cancer cells by biting, and both cause facial tumours. DFT1 and DFT2 tumours are grossly indistinguishable, but can be differentiated using histopathology, cytogenetics or genotyping of polymorphic markers. However, standard diagnostic methods require specialist skills and equipment and entail long processing times. Here, we describe Tasman-PCR: a simple polymerase chain reaction (PCR)-based diagnostic assay that identifies and distinguishes DFT1 and DFT2 by amplification of DNA spanning tumour-specific interchromosomal translocations. We demonstrate the high sensitivity and specificity of this assay by testing DNA from 546 tumours and 804 normal devils. A temporal–spatial screen confirmed the reported geographic ranges of DFT1 and DFT2 and did not provide evidence of additional DFT clones. DFT2 affects disproportionately more males than females, and devils can be co-infected with DFT1 and DFT2. Overall, we present a PCR-based assay that delivers rapid, accurate and high-throughput diagnosis of DFT1 and DFT2. This tool provides an additional resource for devil disease management and may assist with ongoing conservation efforts.

Context Conservation management relies on baseline demographic data of natural populations. For Tasmanian devils (Sarcophilus harrisii), threatened in the wild by two fatal and transmissible cancers (devil facial tumour disease DFTD: DFT1 and DFT2), understanding the characteristics of healthy populations is crucial for developing adaptive management strategies to bolster populations in the wild. Aims Our analysis aims to evaluate contemporary reproductive rates for wild, DFTD-free Tasmanian devil populations, and to provide a baseline with which to compare the outcome of current translocation activities. Methods We analysed 8 years of field-trapping data, including demographics and reproductive rates, across 2004–16, from the largest known DFTD-free remnant population at Woolnorth, Tasmania. Key results Surprisingly, we found a dramatic and statistically significant decline in female breeding rate when comparing data collected from 2004–2009 with data from 2014–2016. Unfortunately we do not have any data from the intermediate years. This decline in breeding rate was accompanied by a subtle but statistically significant decline in litter sizes. These changes were not associated with a change in body condition over the same period. Furthermore, we could not attribute the decline in breeding to a change in population size or sex ratio. Preliminary analysis suggested a possible association between annual breeding rate and coarse measures of environmental variation (Southern Oscillation Index), but any mechanistic associations are yet to be determined. Conclusions The decline in breeding rates was unexpected, so further monitoring and investigation into potential environmental and/or biological reasons for the decline in breeding rate are recommended before the arrival of DFTD at Woolnorth. Implications Our results provide valuable data to support the conservation management of Tasmanian devils in their native range. They also highlight the importance of continued monitoring of ‘safe’ populations, in the face of significant threats elsewhere.

Wildlife roadkill is a worldwide issue being tackled using several mitigation methods that have variable efficacy. Tasmania has a significant roadkill issue, and several endemic species that are vulnerable to roadkill. Virtual fence devices are manufactured in Austria and were originally designed to mitigate against the impact of vehicles hitting large game species such as deer and boar. Here, we trial the devices over three years at a single site in Tasmania to determine whether they are effective in reducing roadkill of Tasmanian mammal species. While there was no spatial replication in this trial, a reduction in total roadkill rate, and in the most commonly affected species, by 50% suggests that these devices have enormous potential to substantially reduce roadkill rates. Many unique mammal species no longer found in the wild on mainland Australia are still found in Tasmania, making mitigating roadkill hotspots an important conservation tool to help maintain presence of these species in their last remaining stronghold. Road managers, including councils, in other Australian States that suffer from high rates of wildlife roadkill may benefit from the knowledge of the results of this trial.

The Tasmanian devil, a marsupial carnivore, has been restricted to the island state of Tasmania since its extinction on the Australian mainland about 3000 years 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 20 years, 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 multiinstitutional 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.

1. Monitoring the response of wild mammal populations to threatening processes is fundamental to effective conservation management. This is especially true for infectious diseases, which may have dynamic and therefore unpredictable interactions with their host. 2. We investigate the long-term impact of a transmissible cancer, devil facial tumour disease (DFTD), on the endemic Tasmanian devil. We analyse trends in devil spot-light counts and density across the area impacted by the disease. We investigate the demographic parameters which might be driving these trends, and use spatial capture-recapture models to examine whether DFTD has affected home range size. 3. We found that devils have declined by an average of 77% in areas affected by DFTD, and that there is a congruent trend of ongoing small decline in spotlight counts and density estimates. Despite this, devils have persisted to date within each of nine monitoring sites. One site is showing as yet unexplained small increases in density 8-10 years after the emergence of DFTD. 4. We also found the prevalence of DFTD has not abated despite large declines in density and that diseased sites continue to be dominated by young devils. The long-term impact of the disease has been partially offset by increased fecundity in the form of precocial breeding in 1-year-old females, and more pouch young per female in diseased sites. The lower densities resulting from DFTD did not affect home range size. 5. Synthesis and applications. Transmission of devil facial tumour disease continues despite large declines in devil density over multiple generations. Plasticity in life history traits has ameliorated the impact of devil facial tumour disease, however broad-scale trends in density show ongoing decline. In light of this, devil facial tumour disease and the impact of stochastic events on the reduced densities wrought by the disease, continue to threaten devils. In the absence of methods to manage disease in wild populations, we advocate managing the low population densities resulting from disease rather than disease per se.