Growth and development of the Tasmanian devil (Sarcophilus harrisii) at Healesville Sanctuary,Victoria, Australia

A study of the growth and development of the Tasmanian devil (Sarcophilus harrisii) was undertaken on a litter born at Healesville Sanctuary in Healesville, Victoria, Australia. Measurements of crown–rump length, head length, and head width were recorded every week from birth. Body weight was recorded weekly once the joeys were detached from the teat at 142 days of age, until 218 days of age. The growth curves revealed an initial linear increase in crown–rump lengths until approximately 60 days of age, when the gradient increased, while head length and head width showed a largely linear increase with age. The appearances of various morphological characteristics were also recorded. Zoo Biol 22:497–505, 2003. © 2003 Wiley‐Liss, Inc.     

Primary productivity and trophic status of Lakes Sorell and Crescent,Tasmania

The primary productivity of two turbid, shallow lakes on the Tasmanian Central Plateau was determined by the C¹⁴ technique from half-light day incubations in situ. Graphical integration of depth-rate curves gave estimates of areal day rates of production and of annual rates.The 2 lakes are closely adjacent and very similar physically and chemically, but have very different phytoplankton populations. Lake Crescent has ten times the standing crop biomass of Lake Sorell but its greater turbidity restricts light penetration, and production per unit of surface per day and per year is only 2.6 times that of Sorell.With day rates of 25-(44)-93 mgCm^–2 and annual production of 16.9 gCm^–2 Lake Sorell could be regarded as oligotrophic. Consideration of standing crop biomass and morphometry however indicates oligo-mesotrophy. Lake Crescent with day rates of 35-(115)-250 mgCm^–2 and annual production of 45 gCm^–2 is moderately eutrophic.Incubations in constant light demonstrated considerable variation in production rates in different parts of Lake Crescent.     

Establishment of a monoclonal antibody PMab-233 for immunohistochemical analysis against Tasmanian devil podoplanin

Monoclonal antibodies (mAbs) against not only human, mouse, and rat but also rabbit, dog, cat, bovine, pig, and horse podoplanins (PDPNs) have been established in our previous studies. PDPN is used as a lymphatic endothelial cell marker in pathological diagnoses. However, mAbs against Tasmanian devil PDPN (tasPDPN), which are useful for immunohistochemical analysis, remain to be developed. Herein, mice were immunized with tasPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/tasPDPN) cells, and hybridomas producing mAbs against tasPDPN were screened using flow cytometry. One of the mAbs, PMab-233 (IgG₁, kappa), specifically detected CHO/tasPDPN cells by flow cytometry and recognized tasPDPN protein by western blotting. Furthermore, PMab-233 strongly detected CHO/tasPDPN cells by immunohistochemistry. These findings suggest that PMab-233 may be useful as a lymphatic endothelial cell marker of the Tasmanian devil.     

Hormonal control of birth behavior in the Tasmanian devil Sarcophilus harrisii

In a number of marsupial species, females exhibit characteristic, stereotyped parturient behavior that facilitates the passage of the neonates to the pouch. In macropodids, this parturient behavior can be induced in non-pregnant females and males by treatment with either prostaglandin F2alpha (PGF2alpha) or oxytocin (OT). This study investigated the effects of PGF2alpha and OT on behavior of Tasmanian devils. Animals tended to sit or lie down quickly, with little vocalization, after treatment with PGF2alpha or OT, while after saline, the animals remained alert, seldom sat, and frequently vocalized. Hormone treatment caused increased respiration. Urogenital and pouch grooming, a characteristic element of parturient behavior in macropodids, was seen in only one devil after hormone treatment. However, no pouch or urogenital grooming was seen in videotape of a devil giving birth, so this may not be a feature of parturient behavior in this species. Overall behavior of males and females was very similar suggesting that the behavioral effects observed may be due to direct neural action of PGF2alpha or OT, rather than an indirect response to uterine or vaginal contractions caused by the hormones. This study is the first to demonstrate that OT results in PGF2alpha secretion as PGFM levels rose after OT injection.     

Contagious cancer: lessons from the devil and the dog

Cancer is generally defined as uncontrollable growth of cells caused by genetic aberrations and/or environmental factors. Yet contagious cancers also occur. The recent emergence of a contagious cancer in Tasmanian devils has reignited interest in transmissible cancers. Two naturally occurring transmissible cancers are known: devil facial tumour disease and canine transmissible venereal tumour. Both cancers evolved once and have then been transmitted from one individual to another as clonal cell lines. The dog cancer is ancient; having evolved more than 6,000 years ago, while the devil disease was first seen in 1996. In this review I will compare and contrast the two diseases focusing on the life histories of the clonal cell lines, their evolutionary trajectories and the mechanisms by which they have achieved immune tolerance. A greater understanding of these contagious cancers will provide unique insights into the role of the immune system in shaping tumour evolution and may uncover novel approaches for treating human cancer.     

Characterization of major histocompatibility complex class I and class II genes from the Tasmanian devil (<Emphasis Type="Italic">Sarcophilus harrisii</Emphasis>)

The Tasmanian devil (Sarcophilus harrisii) is currently threatened by an emerging wildlife disease, devil facial tumour disease. The disease is decreasing devil numbers dramatically and may lead to the extinction of the species. At present, nothing is known about the immune genes or basic immunology of the devil. In this study, we report the construction of the first genetic library for the Tasmanian devil, a spleen cDNA library, and the isolation of full-length MHC Class I and Class II genes. We describe six unique Class II beta chain sequences from at least three loci, which belong to the marsupial Class II DA gene family. We have isolated 13 unique devil Class I sequences, representing at least seven Class I loci, two of which are most likely non-classical genes. The MHC Class I sequences from the devil have little heterogeneity, indicating recent divergence. The MHC genes described here are most likely involved in antigen presentation and are an important first step for studying MHC diversity and immune response in the devil.     

A numerical analysis of Tasmanian higher plant endemism

KIRKPATRICK, J. B. & BROWN, M. J., 1984. A numerical analysis of Tasmanian higher plant endemism. Tasmanian endemic plant taxa at the species level or below were placed in geographic elements according to the distribution of their genera. These elements are associated with different environments, the endemic and Antarctic elements being most prominent in rainforest and alpine communities; the cosmopolitan element in alpine communities, and the Australian element in the fire-prone lowland communities. A monothetic divisive classification and minimum spanning tree analysis of a matrix of the occurrence of 242 Tasmanian endemic species from 366 areas resulted in groups of species and lists whose distributions were closely related to precipitation, altitude and vegetation type. The proportions of endemic species in local Tasmanian floras were almost totally explained by altitude and precipitation in a stepwise multiple regression analysis. However, it is possible that many of the endemic species have not been able to occupy their potential range in Tasmania as a result of insufficient time having elapsed for them to fully expand from Glacial refugia.     

Increasing turbidity significantly alters the diet of brown trout: a multi‐year longitudinal study

The stomach contents of adult brown trout Salmo trutta from Lake Sorell, Tasmania, were analysed during 6 years of increasing turbidity to follow changes in the diet associated with dramatic changes in water clarity. Brown trout were sampled from 1996, when turbidity was 26 NTU, to 2001 when turbidity was 141 NTU. The mean relative volume of stomach contents decreased progressively to 2001, by which time it was only one sixth of that in 1996, and the mean diversity of prey in stomachs decreased from an average of more than six species per stomach in 1996 to one species in 2001. The species composition of stomach contents shifted from domination by the phreatoicid isopod Colubotelson sp., to the galaxiid fish Galaxias auratus and the amphipod Austrochiltonia australis , and then the cladoceran Daphnia carinata . To give an indication of diet changes over a typical yearly cycle in the current turbid state of the lake, a sample was taken from each season from December 2000 to September 2001. Two basic diets were found during the year; brown trout specialized on D. carinata in summer and autumn, and G. auratus in winter and spring. Mean diversity of prey was less than two species per stomach in all samples from 2000 to 2001, except for the sample from spring 2001 when it was 2·2 species per stomach, and the mean relative volume of stomach contents was more than three times greater in winter than any other season. The ways in which high turbidity may have influenced the changes in the brown trout diet observed since 1996 and the patterns evident during the seasons of 2000–2001 are discussed.     

Distribution and Impacts of Tasmanian Devil Facial Tumor Disease

The Tasmanian devil, Sarcophilus harrisii, is the largest extant marsupial carnivore. In 1996, a debilitating facial tumor was reported. It is now clear that this is an invariably lethal infectious cancer. The disease has now spread across the majority of the range of the species and is likely to occur across the entire range within 5 to 10 years. The disease has lead to continuing declines of up to 90% and virtual disappearance of older age classes. Mark-recapture analysis and a preliminary epidemiological model developed for the population with the best longitudinal data both project local extinction in that area over a timeframe of 10 to 15 years from disease emergence. However, the prediction of extinction from the model is sensitive to the estimate of the latent period, which is poorly known. As transmission appears to occur by biting, much of which happens during sexual encounters, the dynamics of the disease may be typical of sexually transmitted diseases. This means that transmission is likely to be frequency-dependent with no threshold density for disease maintenance. Extinction over the entire current range of the devil is therefore a real possibility and an unacceptable risk.     

MHC gene copy number variation in Tasmanian devils: implications for the spread of a contagious cancer

Tasmanian devils face extinction owing to the emergence of a contagious cancer. Devil facial tumour disease (DFTD) is a clonal cancer spread owing to a lack of major histocompatibility complex (MHC) barriers in Tasmanian devil populations. We present a comprehensive screen of MHC diversity in devils and identify 25 MHC types and 53 novel sequences, but conclude that overall levels of MHC diversity at the sequence level are low. The majority of MHC Class I variation can be explained by allelic copy number variation with two to seven sequence variants identified per individual. MHC sequences are divided into two distinct groups based on sequence similarity. DFTD cells and most devils have sequences from both groups. Twenty per cent of individuals have a restricted MHC repertoire and contain only group I or only group II sequences. Counterintuitively, we postulate that the immune system of individuals with a restricted MHC repertoire may recognize foreign MHC antigens on the surface of the DFTD cell. The implication of these results for management of DFTD and this endangered species are discussed.     

Extensive population decline in the Tasmanian devil predates European settlement and devil facial tumour disease

The Tasmanian devil (Sarcophilus harrisii) was widespread in Australia during the Late Pleistocene but is now endemic to the island of Tasmania. Low genetic diversity combined with the spread of devil facial tumour disease have raised concerns for the species’ long-term survival. Here, we investigate the origin of low genetic diversity by inferring the species'' demographic history using temporal sampling with summary statistics, full-likelihood and approximate Bayesian computation methods. Our results show extensive population declines across Tasmania correlating with environmental changes around the last glacial maximum and following unstable climate related to increased ‘El Niño–Southern Oscillation’ activity.     

Microsatellites for the Tasmanian devil (Sarcophilus laniarius)

The Tasmanian devil (Sarcophilus laniarius), a medium‐sized predator/scavenger, is the largest member of the short‐lived carnivorous marsupial Family Dasyuridae. Now restricted to Tasmania, populations are impacted by habitat clearance and anthropogenic mortality and genetic studies could be of value in informing levels of genetic diversity, mating system, dispersal and the effects of natural and anthropogenic landscape features on gene flow. Microsatellite markers were isolated from a partial, size‐selected genomic library that was enriched for microsatellite sequences. Primer pairs were developed for 11 polymorphic dinucleotide microsatellite loci that conform with Hardy–Weinberg equilibrium and reveal moderate genetic variability across the species range.     

Reproductive biology of the threatened golden galaxias Galaxias auratus Johnston and the influence of lake hydrology

Golden galaxias Galaxias auratus (31–235 mm fork length, L _F) were collected monthly from littoral habitats in Lakes Crescent and Sorell, Tasmania, Australia, between July 2000 and December 2002. Spawning habitats were identified and monitored in both lakes, and surveyed in Lake Crescent. Trends in gonado-somatic indices and reproductive stages of development indicated that gonad development in both sexes begins in midsummer and peaks in late autumn to early winter. Males mature at smaller sizes (50% at 52 mm L _F) than females (50% at 76 mm L _F), larger individuals are predominately females (95% of fish ≥138 mm L _F), and overall male to female ratios are female biased ( c . 1:2). Spawning occurs late autumn to early spring (water temperatures = 1·4–9·7° C) with peaks in spawning activity in winter (mean water temperatures <5° C). Demersal adhesive eggs ( c. 1·5 mm diameter) were found on cobble substrata ( c. 20–250 mm diameter) in littoral areas ( c. 0·2–0·6 m deep) and fecundity of fish 71–181 mm L _F ranged from 619 to 14 478 eggs. The rate of change in water level over the 20 days prior to monthly sampling was important in explaining the occurrence of spent fish and this accounted for temporal differences in spawning between the populations. Lake hydrology influences the reproductive cycle of G. auratus by possibly providing a stimulus for spawning and it controls the availability of spawning habitat in Lake Crescent. Seasonal hydrological cycles ( i.e. rises during late autumn to winter) and a minimum water level of 802·20 m Australian Height Datum in Lake Crescent during autumn (above which littoral areas of cobble substratum are inundated) are critical to G. auratus populations.     

Role of foliar chemistry versus leaf-tip morphology in egg-batch placement by Chrysophtharta bimaculata (Olivier) (Coleoptera: Chrysomelidae)

Abstract The Tasmanian eucalyptus leaf beetle, Chrysophtharta bimaculata (Olivier) (Chrysomelidae: Paropsini), deposits batches of eggs on the leaf surfaces of its Eucalyptus hosts. Using a primary host of the beetle, Eucalyptus regnans , the aim of the present study was to examine whether oviposition site discrimination was occurring at the level of the individual leaf and what, if any, chemical attributes were involved in site selection. From field surveys, it was found that 55−65% of egg batches were laid in the leaf-tip zone, despite this zone representing only 5.6% of the total leaf-surface area. Chemical analyses of expanding leaves showed that the concentrations of three essential oil components and three surface waxes varied significantly between the leaf tip and remainder of the leaf. However, on plastic mimic leaves with uniform, non-foliar chemistry, eggs were still preferentially laid near 'leaf' tips. Hence, leaf essential oils and surface waxes need not be involved in oviposition site choice. Observations of ovipositing beetles showed that they wrapped their tarsi around the leaf margin, suggesting that the ability to grasp the edges of a leaf may explain the oviposition pattern observed.     

The impact of disease on the survival and population growth rate of the Tasmanian devil

1. We investigated the impact of a recently emerged disease, Devil Facial Tumour Disease (DFTD), on the survival and population growth rate of a population of Tasmanian devils, Sarcophilus harrisii, on the Freycinet Peninsula in eastern Tasmania. 2. Cormack-Jolly-Seber and multistate mark-recapture models were employed to investigate the impact of DFTD on age- and sex-specific apparent survival and transition rates. Disease impact on population growth rate was investigated using reverse-time mark-recapture models. 3. The arrival of DFTD triggered an immediate and steady decline in apparent survival rates of adults and subadults, the rate of which was predicted well by the increase in disease prevalence in the population over time. 4. Transitions from healthy to diseased state increased with disease prevalence suggesting that the force of infection in the population is increasing and that the epidemic is not subsiding. 5. The arrival of DFTD coincided with a marked, ongoing decline in the population growth rate of the previously stable population, which to date has not been offset by population compensatory responses.     

Evidence that disease-induced population decline changes genetic structure and alters dispersal patterns in the Tasmanian devil

Infectious disease has been shown to be a major cause of population declines in wild animals. However, there remains little empirical evidence on the genetic consequences of disease-mediated population declines, or how such perturbations might affect demographic processes such as dispersal. Devil facial tumour disease (DFTD) has resulted in the rapid decline of the Tasmanian devil, Sarcophilus harrisii, and threatens to cause extinction. Using 10 microsatellite DNA markers, we compared genetic diversity and structure before and after DFTD outbreaks in three Tasmanian devil populations to assess the genetic consequences of disease-induced population decline. We also used both genetic and demographic data to investigate dispersal patterns in Tasmanian devils along the east coast of Tasmania. We observed a significant increase in inbreeding (F_IS pre/post-disease −0.030/0.012, P<0.05; relatedness pre/post-disease 0.011/0.038, P=0.06) in devil populations after just 2–3 generations of disease arrival, but no detectable change in genetic diversity. Furthermore, although there was no subdivision apparent among pre-disease populations (θ=0.005, 95% confidence interval (CI) −0.003 to 0.017), we found significant genetic differentiation among populations post-disease (θ=0.020, 0.010–0.027), apparently driven by a combination of selection and altered dispersal patterns of females in disease-affected populations. We also show that dispersal is male-biased in devils and that dispersal distances follow a typical leptokurtic distribution. Our results show that disease can result in genetic and demographic changes in host populations over few generations and short time scales. Ongoing management of Tasmanian devils must now attempt to maintain genetic variability in this species through actions designed to reverse the detrimental effects of inbreeding and subdivision in disease-affected populations.     

Technique for washing nectar from the flowers of Tasmanian leatherwood (Eucryphia lucida Eucryphiaceae)

The technique for extracting floral nectar using micropipettes is often unsuited to flowers where nectar is produced in very small volumes and/or where nectar is highly viscous. An alternative technique for washing the viscous nectar from the flowers of Tasmanian leatherwood, Eucryphia lucida (Labill.) is described. Here, two washes with a known volume of distilled water removed 95% of total floral sugar. Using such a washing technique on exposed and bagged E. lucida flowers provided information on nectar production in this species that could not have been obtained using a standard micropipette extraction method.     

Lack of genetic diversity across diverse immune genes in an endangered mammal,the Tasmanian devil (Sarcophilus harrisii)

The Tasmanian devil (Sarcophilus harrisii) is threatened with extinction due to the spread of devil facial tumour disease. Polymorphisms in immune genes can provide adaptive potential to resist diseases. Previous studies in diversity at immune loci in wild species have almost exclusively focused on genes of the major histocompatibility complex (MHC); however, these genes only account for a fraction of immune gene diversity. Devils lack diversity at functionally important immunity loci, including MHC and Toll‐like receptor genes. Whether there are polymorphisms at devil immune genes outside these two families is unknown. Here, we identify polymorphisms in a wide range of key immune genes, and develop assays to type single nucleotide polymorphisms (SNPs) within a subset of these genes. A total of 167 immune genes were examined, including cytokines, chemokines and natural killer cell receptors. Using genome‐level data from ten devils, SNPs within coding regions, introns and 10 kb flanking genes of interest were identified. We found low polymorphism across 167 immune genes examined bioinformatically using whole‐genome data. From this data, we developed long amplicon assays to target nine genes. These amplicons were sequenced in 29–220 devils and found to contain 78 SNPs, including eight SNPS within exons. Despite the extreme paucity of genetic diversity within these genes, signatures of balancing selection were exhibited by one chemokine gene, suggesting that remaining diversity may hold adaptive potential. The low functional diversity may leave devils highly vulnerable to infectious disease, and therefore, monitoring and preserving remaining diversity will be critical for the long‐term management of this species. Examining genetic variation in diverse immune genes should be a priority for threatened wildlife species. This study can act as a model for broad‐scale immunogenetic diversity analysis in threatened species.