Captive Devils Research Articles

Generation of Devil Facial Tumour Cells Co-Expressing MHC With CD80, CD86 or 41BBL to Enhance Tumour Immunogenicity.

The major histocompatibility complex (MHC) molecules play an integral role in the adaptive immune response to transmissible cancers through tumour antigen presentation and recognition of allogeneic MHC molecules. The transmissible devil facial tumours 1 and 2 (DFT1 and DFT2) modulate MHC-I antigen presentation to evade host immune responses and facilitate transmission of tumours cells to new Tasmanian devil (Sarcophilus harrisii) hosts. To enhance T-cell-driven tumour immunogenicity for vaccination and immunotherapy, DFT1 and DFT2 cells were co-transfected with (i) NLRC5 for MHC-I expression or CIITA for MHC-I and MHC-II expression, and (ii) a co-stimulatory molecule, either CD80, CD86 or 41BBL. The co-transfected DFT cells presented enhanced expression of MHC-I and/or MHC-II. As few devil-specific monoclonal antibodies exist, we used recombinant CTLA4 and 41BB fused to a fluorescent protein to confirm expression of cell surface CD80, CD86 and 41BBL. The capacity for these cells to induce T-cell responses including PD1 and IFNG expression was evaluated in in vitro co-culture assays with captive devil peripheral blood mononuclear cells (PBMCs). Although PBMC viability had increased, there was no evidence of enhanced T-cell activation. This system can be used to identify additional factors required to promote activation of naïve devil T-cells in vitro.

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.

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.

Post-release immune responses of Tasmanian devils vaccinated with an experimental devil facial tumour disease vaccine

Abstract Context Disease is increasingly becoming a driver of wildlife population declines and an extinction risk. Vaccines are 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. In 2016, 33 devils from a DFTD-free insurance population were given an experimental DFTD vaccination before their wild release on the Tasmanian northern coast. Aim To determine the efficacy of the vaccination protocol and the longevity of the induced responses. Method Six trapping trips took place over the 2.5 years following release, and both vaccinated and incumbent devils had blood samples and tumour biopsies collected. Key results In all, 8 of the 33 vaccinated devils were re-trapped, and six of those developed DFTD within the monitoring period. Despite the lack of protection provided by the vaccine, we observed signs of immune activation not usually found in unvaccinated devils. First, sera collected from the eight devils showed that anti-DFTD antibodies persisted for up to 2 years post-vaccination. Second, tumour-infiltrating lymphocytes were found in three of four biopsies collected from vaccinated devils, which contrasts with the ‘immune deserts’ typical of DFTs; only 1 of the 20 incumbent devils with DFTD had a tumour biopsy exhibiting immune-cell infiltrate. Third, immunohistochemical analysis of the vaccinated devils’ tumour biopsies identified the functional immune molecules associated with antigen-presenting cells (MHC-II) and T-cells (CD3), and the immune checkpoint molecule PD-1, all being associated with anti-tumour immunity in other species. Conclusions 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. Implications A protective DFTD vaccine would provide a valuable management approach for conservation of the Tasmanian devil.

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.

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.

MHC diversity and female age underpin reproductive success in an Australian icon; the Tasmanian Devil

Devil Facial Tumour Disease (DFTD), a highly contagious cancer, has decimated Tasmanian devil (Sarcophilus harrisii) numbers in the wild. To ensure its long-term survival, a captive breeding program was implemented but has not been as successful as envisaged at its launch in 2005. We therefore investigated the reproductive success of 65 captive devil pair combinations, of which 35 produced offspring (successful pairs) whereas the remaining 30 pairs, despite being observed mating, produced no offspring (unsuccessful pairs). The devils were screened at six MHC Class I-linked microsatellite loci. Our analyses revealed that younger females had a higher probability of being successful than older females. In the successful pairs we also observed a higher difference in total number of heterozygous loci, i.e. when one devil had a high total number of heterozygous loci, its partner had low numbers. Our results therefore suggest that devil reproductive success is subject to disruptive MHC selection, which to our knowledge has never been recorded in any vertebrate. In order to enhance the success of the captive breeding program the results from the present study show the importance of using young (2-year old) females as well as subjecting the devils to MHC genotyping.

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.

The Tasmanian devil microbiome-implications for conservation and management.

BackgroundThe Tasmanian devil, the world’s largest carnivorous marsupial, is at risk of extinction due to devil facial tumour disease (DFTD), a fatal contagious cancer. The Save the Tasmanian Devil Program has established an insurance population, which currently holds over 600 devils in captive facilities across Australia. Microbes are known to play a crucial role in the health and well-being of humans and other animals, and increasing evidence suggests that changes in the microbiota can influence various aspects of host physiology and development. To improve our understanding of devils and facilitate management and conservation of the species, we characterised the microbiome of wild devils and investigated differences in the composition of microbial community between captive and wild individuals.ResultsA total of 1,223,550 bacterial 16S ribosomal RNA (rRNA) sequences were generated via Roche 454 sequencing from 56 samples, including 17 gut, 15 skin, 18 pouch and 6 oral samples. The devil’s gut microbiome was dominated by Firmicutes and showed a high Firmicutes-to-Bacteroidetes ratio, which appears to be a common feature of many carnivorous mammals. Metabolisms of carbohydrates, amino acids, energy, cofactors and vitamins, nucleotides and lipids were predicted as the most prominent metabolic pathways that the devil's gut flora contributed to. The microbiota inside the female’s pouch outside lactation was highly similar to that of the skin, both co-dominated by Firmicutes and Proteobacteria. The oral microbiome had similar proportions of Proteobacteria, Bacteroidetes, Firmicutes and Fusobacteria.ConclusionsCompositional differences were observed in all four types of microbiota between devils from captive and wild populations. Certain captive devils had significantly lower levels of gut bacterial diversity than wild individuals, and the two groups differed in the proportion of gut bacteria accounting for the metabolism of glycan, amino acids and cofactors and vitamins. Further studies are underway to investigate whether alterations in the microbiome of captive devils can have impacts on their ability to adapt and survive following re-introduction to the wild.Electronic supplementary materialThe online version of this article (doi:10.1186/s40168-015-0143-0) contains supplementary material, which is available to authorized users.

A preliminary study assessing risk to Tasmanian devils from poisoning for red foxes

AbstractThe recent introduction of red foxes (Vulpes vulpes) to Australia's island state of Tasmania represents a major threat to native fauna. In response, the Tasmanian government has begun a fox eradication program using Foxoff®, a bait containing the poison sodium monofluoroacetate (commonly known as 1080). The bait is potentially attractive to native Tasmanian carnivores as well as to foxes. Of particular concern is the endangered Tasmanian devil (Sarcophilus harrisii), which is already at risk from an emergent infectious disease, devil facial tumor disease (DFTD). In both a captive and a field study using non‐toxic Foxoff bait, we assessed bait palatability and possible effects of demographics, hunger level, bait age, and bait burial method on the likelihood of bait uptake by Tasmanian devils. Captive devils showed varying interest in the bait, but wild devils appeared to find it uniformly palatable. In the captive study, males and younger, captive‐born animals were more likely to excavate and remove bait. Subterranean burial at 15 cm was the most effective deterrent to bait excavation; effectiveness decreased at shallower depths and with surface‐level bait buried beneath soil mounds. Our findings suggest that the current fox‐baiting campaign may negatively impact individual devils. More extensive study is necessary to assess potential risk at the population level. © 2011 The Wildlife Society.

The diet of the Tasmanian Devil, Sarcophilus harrisii, as determined from analysis of scat and stomach contents

Knowledge of the diets of carnivores is an essential precursor to understanding their role as predators in ecosystems. To date, understanding of the diet of Tasmanian Devils, Sarcophilus harrisii, is limited and based upon largely qualitative descriptions. We examined the diets of Tasmanian Devils at six sites by identifying undigested hair, bone and feathers found in their scats. These sites range across different habitat types in coastal and inland Tasmania, and encompass devil populations that are known as both free of the Devil Facial Tumour Disease (DFTD) and populations that are infected by the disease. Tasmanian Devil scats at coastal sites (n=27) contained ten species of mammal, as well as birds, fish and insects. Scats collected from inland sites (n= 17) were comprised of six mammalian species, birds and invertebrates. The most common food items were birds, Common Brushtail and Ringtail possums (Trichosurus vulpecula and Pseudocheirus peregrinus respectively), Tasmanian Pademelons (Thylogale billardierii) and Bennett's Wallabies (Macropus ruftgriseus). O fall the scats, 61% contained only one food group, 32% contained two groups, 4% contained three food items and only one scat (2%) contained four food groups. We supplement this information with stomach contents from road-killed devils, and compare our results with those of previous studies, with a view to furthering our understanding ofthe ecology ofthe threatened Tasmanian Devil. Such information will be important for the management of wild and captive devil populations, particularly in light of DFTD.

Habitat and Substrate Use in Reproduction of Captive Devils River Minnows

AbstractThe Devils River minnow Dionda diaboli is a threatened species once endemic to a portion of the Rio Grande drainage in western Texas and northern Mexico but now found only in the Devils River and San Felipe and Pinto creeks. Because little is known about the biology of this species, it is difficult to establish protocols for maintaining captive populations. We monitored the production of Devils River minnows in two laboratory culture systems containing four substrate types (rocks, gravel, sand, and Spawntex) within each of two habitat types (riffle and pool) that had two subhabitat types each (upper and lower riffle, covered and uncovered pool). A total of 38 adult fish (mean weight = 1.38 g; mean total length = 54 mm) were introduced into the two culture systems (19 fish each, approximately equal sex ratio) on 5 September 2001. From 18 September to 6 December, 2,269 young were removed. A significantly greater number (1,922; three‐factor nested analysis of variance [ANOVA], Fisher's least‐significant‐difference tests, P ≤ 0.05) were in gravel substrate (2–5 cm diameter). However, no differences were found in the counts of young with respect to habitat or subhabitat type. The indoor culture of Devils River minnows is possible using gravel as a spawning substrate and, since parental stock can be manipulated, the technique can be used for establishing refugium populations and for augmenting wild populations.