A Prototype Recombinant-Protein Based Chlamydia pecorum Vaccine Results in Reduced Chlamydial Burden and Less Clinical Disease in Free-Ranging Koalas (Phascolarctos cinereus)

Diseases associated with Chlamydia pecorum infection are a major cause of decline in koala populations in Australia. While koalas in care can generally be treated, a vaccine is considered the only option to effectively reduce the threat of infection and disease at the population level. In the current study, we vaccinated 30 free-ranging koalas with a prototype Chlamydia pecorum vaccine consisting of a recombinant chlamydial MOMP adjuvanted with an immune stimulating complex. An additional cohort of 30 animals did not receive any vaccine and acted as comparison controls. Animals accepted into this study were either uninfected (Chlamydia PCR negative) at time of initial vaccination, or infected (C. pecorum positive) at either urogenital (UGT) and/or ocular sites (Oc), but with no clinical signs of chlamydial disease. All koalas were vaccinated / sampled and then re-released into their natural habitat before re-capturing and re-sampling at 6 and 12 months. All vaccinated koalas produced a strong immune response to the vaccine, as indicated by high titres of specific plasma antibodies. The incidence of new infections in vaccinated koalas over the 12-month period post-vaccination was slightly less than koalas in the control group, however, this was not statistically significant. Importantly though, the vaccine was able to significantly reduce the infectious load in animals that were Chlamydia positive at the time of vaccination. This effect was evident at both the Oc and UGT sites and was stronger at 6 months than at 12 months post-vaccination. Finally, the vaccine was also able to reduce the number of animals that progressed to disease during the 12-month period. While the sample sizes were small (statistically speaking), results were nonetheless striking. This study highlights the potential for successful development of a Chlamydia vaccine for koalas in a wild setting.     

Inbreeding and disease avoidance in a free‐ranging koala population

Habitat destruction and fragmentation are increasing globally, forcing surviving species into small, isolated populations. Isolated populations typically experience heightened inbreeding risk and associated inbreeding depression and population decline; although individuals in these populations may mitigate these risks through inbreeding avoidance strategies. For koalas, as dietary specialists already under threat in the northern parts of their range, increased habitat fragmentation and associated inbreeding costs are of great conservation concern. Koalas are known to display passive inbreeding avoidance through sex‐biased dispersal, although population isolation will reduce dispersal pathways. We tested whether free‐ranging koalas display active inbreeding avoidance behaviours. We used VHF tracking data, parentage reconstruction, and veterinary examination results to test whether free‐ranging female koalas avoid mating with (a) more closely related males; and (b) males infected with sexually transmitted Chlamydia pecorum. We found no evidence that female koalas avoid mating with relatively more related available mates. In fact, as the relatedness of potential mates increases, so did inbreeding events. We also found no evidence that female koalas can avoid mating with males infected with C. pecorum. The absence of active inbreeding avoidance mechanisms in koalas is concerning from a conservation perspective, as small, isolated populations may be at even higher risk of inbreeding depression than expected. At risk koala populations may require urgent conservation interventions to augment gene flow and reduce inbreeding risks. Similarly, if koalas are not avoiding mating with individuals with chlamydial disease, populations may be at higher risk from disease than anticipated, further impacting population viability.     

New technologies in the mix: Assessing N‐mixture models for abundance estimation using automated detection data from drone surveys

1. Reliable estimates of abundance are critical in effectively managing threatened species, but the feasibility of integrating data from wildlife surveys completed using advanced technologies such as remotely piloted aircraft systems (RPAS) and machine learning into abundance estimation methods such as N‐mixture modeling is largely unknown due to the unique sources of detection errors associated with these technologies.
2. We evaluated two modeling approaches for estimating the abundance of koalas detected automatically in RPAS imagery: (a) a generalized N‐mixture model and (b) a modified Horvitz–Thompson (H‐T) estimator method combining generalized linear models and generalized additive models for overall probability of detection, false detection, and duplicate detection. The final estimates from each model were compared to the true number of koalas present as determined by telemetry‐assisted ground surveys.
3. The modified H‐T estimator approach performed best, with the true count of koalas captured within the 95% confidence intervals around the abundance estimates in all 4 surveys in the testing dataset (n = 138 detected objects), a particularly strong result given the difficulty in attaining accuracy found with previous methods.
4. The results suggested that N‐mixture models in their current form may not be the most appropriate approach to estimating the abundance of wildlife detected in RPAS surveys with automated detection, and accurate estimates could be made with approaches that account for spurious detections.

     

A comparison and critique of different scat‐analysis methods for determining carnivore diet

• 1 For terrestrial carnivores, scat analysis is the technique most often used to determine diets. Various methods of interpreting scat‐analysis data exist; however, little is known about how the choice of method affects the results.
• 2 We reviewed 50 scat‐analysis papers to assess the range of methods currently used. Furthermore, we used a large data set from cape fox Vulpes chama and black‐backed jackal Canis mesomelas scats to compare 11 scat‐analysis methods. Techniques tested included five biomass calculation methods, four frequency of occurrence methods, one method that estimated volume in scats, and another that estimated mass of food items in scats.
• 3 Frequency of occurrence methods were used in 94% of reviewed papers, and in 50% of papers they were the sole methods used. However, we conclude that frequency of occurrence has the least ecological significance and results can be misleading. Although biomass calculations probably provide the best approximation to true diets, only 23% of reviewed papers used suitable biomass calculation methods when models were available for the study species.
• 4 Analysis of fox and jackal scats showed that there were significant differences among methods when calculating percent diet composition and niche breadth. Additionally, dietary overlap between species differed considerably among the methods (range of R₀ = 0.29–0.79). We conclude that the choice of method can have a significant impact on the results of dietary analysis, and can lead to very different conclusions about a species' ecology.
• 5 The best approximation of the true diet can be obtained by using a biomass calculation model that was developed for the same species, or for a closely related species with a similar food spectrum. When no such model is available, either the volume or mass of diet components in the scats should be used. To document rare food items, frequency of occurrence data could also be given.

     

Disappearance rate of chimpanzee scats: Implications for census work on Pan troglodytes

Scat (faeces) decay rate estimates are used to calculate animal species abundance and density. For African great apes, this has been measured only for Gorilla; chimpanzee scats are assumed to decay at a faster rate due to lower fibre content. We provide the first systematic measure of scat decay rate duration for Pan troglodytes schweinfurthii, in Kanyawara, Kibale National Park, Uganda. We used two methods: (1) multiple visits to obtain prospective decay rates (PDR) (N = 96 scats) and (2) a novel approach of time‐lapse photography (TLP) (N = 17 scats). Most of the visited scats (67%) decayed in ≤24 hr, and median decay rate duration from photographic documentation was 18 hr. Using regression analyses, we tested 11 covariables to determine predictors for decay rate duration. Greater volume of scat and reduced levels of diurnal dung beetle activity were positively associated with longer decay rate duration. Given a high prevalence of dung beetle activity (88% of scats), particularly within 3 hr post‐defaecation, we suggest the use of the alternative term, disappearance rate of scats. With a rapid disappearance rate, scat count surveys of unhabituated chimpanzees are challenging; further work is then needed for Pan spp. to determine spatial and temporal differences at intra‐ and inter‐species level.     

Diets of sympatric native and introduced carnivores in the Barrington Tops,eastern Australia

Invasive predators have severe impacts on global biodiversity, and their effects in Australia have been more extreme than on any other continent. The spotted‐tailed quoll (Dasyurus maculatus), an endangered marsupial carnivore, coexists with three eutherian carnivores, the red fox (Vulpes vulpes), feral cat (Felis catus) and wild dog (Canis lupus ssp.) with which it did not coevolve. No previous study has investigated dietary overlap between quolls and the suite of three eutherian carnivores. By analysing scats, we aimed to quantify dietary overlap within this carnivore assemblage in eastern Australia, and to detect any differences that may facilitate coexistence. We also sought evidence of intraguild predation. Dietary overlap between predators was extensive, with the greatest similarity occurring between foxes and cats. However, some differences were apparent. For example, cats mainly consumed smaller prey, and wild dogs larger prey. Quolls showed greater dietary overlap with foxes and cats than with dogs. Intraguild predation was evident, with fox remains occurring in 3% of wild dog scats. Our results suggest wild dogs competitively dominate invasive foxes, which in turn are likely to compete with the endangered quoll.     

Using Scent-Matching Dogs to Identify Individual Amur Tigers from Scats

Abstract: Scent-matching dogs have previously been used to identify caged individual Amur tigers (Panthera tigris altaica) by fecal material (scat), but this technique has not been tested in the wild. We tested the hypothesis that trained dogs can identify individual tigers by unique characteristics present in scat. To conduct this work, we used 5 dogs and 58 scats from 25 known individual tigers in independent trials. Dogs correctly selected matched test scats at an average rate of 87% (SE ± 1.4%, n = 521 trials). The average accuracy rates for 4 dogs increased to 98% (SE ± 1.6%, n = 86 sets of repeated-trials) using repeated-trial tests. Each of 5 dogs made correct choices better than expected by chance (dog 1 χ²₁ = 507.9, P ≤ 0.001; dog 2 χ²₁ = 882.1, P ≤ 0.001; dog 3 χ²₁ = 374.1, P ≤ 0.001; dog 4 χ²₁ = 379.2, P ≤ 0.001; and dog 5 χ²₁ = 103.9, P ≤ 0.001). Four dogs were able to match 11 scats deposited over a 4-year period from one tiger with an accuracy of 100% (n = 40 trails). This method may be a useful alternative to genetic analyses that are used in conjunction with scat-sampling schemes in studies for which DNA genotyping is impractical or ineffective. Used with mark—recapture surveys to estimate species abundance, scent-matching dogs have the potential of being important tools in the study of wild Amur tigers, as well as other wildlife species.     

The accuracy of scat identification in distribution surveys: American mink, Neovison vison, in the northern highlands of Scotland

Distribution data for elusive species are often based on detection of field signs rather than of the animal itself. However, identifying field signs can be problematic. We present here the results of a survey for American mink, Neovison vison, in the northern highlands of Scotland to demonstrate the importance of verifying field sign identification. Three experienced surveyors located scats, which they identified as mink scats, at seven of 147 sites surveyed and “possible” mink scats at a further 50 sites. Mitochondrial DNA was successfully extracted from 45 of 75 (60%) scats, collected from 31 of the 57 “positive” sites; sequencing of amplified DNA fragments showed that none of these scats was actually of mink origin. We consider the implications of erroneous survey data and the potential waste of resources and misdirection of conservation/management actions. We discuss potential methods that may be useful to verify field sign identification, including the use of DNA analysis, and stress that verification is crucial to ensure rigorous and reliable survey data.     

Upper respiratory tract disease in captive orangutans (Pongo sp.): prevalence in 20 European zoos and predisposing factors

Background Upper respiratory tract disease (URTD) is a significant cause of morbidity in captive orangutans (Pongo abelii, Pongo pygmaeus), and the pathogenesis is often unknown. Methods The prevalence of respiratory disease in captive European orangutans (201 animals; 20 zoos) and possible predisposing factors were investigated. Results Bornean orangutans (P. pygmaeus) showed chronic respiratory signs significantly more often (13.8%) than Sumatran (P. abelii; 3.6%), and males (15.8%) were more often afflicted than females (3.9%). Hand‐reared animals (21%) developed air sacculitis more often than parent‐reared animals (5%). Diseased animals were more often genetically related to animals with respiratory diseases (93%) than to healthy animals (54%). None of the environmental conditions investigated had a significant effect on disease prevalence. Conclusion Results suggest a higher importance of individual factors for the development of URTD than environmental conditions. Bornean, male and hand‐reared orangutans and animals related to diseased animals need increased medical surveillance for early detection of respiratory disease.     

Fresh is best: Accurate SNP genotyping from koala scats

Maintaining genetic diversity is a crucial component in conserving threatened species. For the iconic Australian koala, there is little genetic information on wild populations that is not either skewed by biased sampling methods (e.g., sampling effort skewed toward urban areas) or of limited usefulness due to low numbers of microsatellites used. The ability to genotype DNA extracted from koala scats using next‐generation sequencing technology will not only help resolve location sample bias but also improve the accuracy and scope of genetic analyses (e.g., neutral vs. adaptive genetic diversity, inbreeding, and effective population size). Here, we present the successful SNP genotyping (1272 SNP loci) of koala DNA extracted from scat, using a proprietary DArTseq^? protocol. We compare genotype results from two‐day‐old scat DNA and 14‐day‐old scat DNA to a blood DNA template, to test accuracy of scat genotyping. We find that DNA from fresher scat results in fewer loci with missing information than DNA from older scat; however, 14‐day‐old scat can still provide useful genetic information, depending on the research question. We also find that a subset of 209 conserved loci can accurately identify individual koalas, even from older scat samples. In addition, we find that DNA sequences identified from scat samples through the DArTseq^? process can provide genetic identification of koala diet species, bacterial and viral pathogens, and parasitic organisms.     

Aspects of the epidemiology of Chlamydia psittaci infection in a population of koalas (Phascolarctos cinereus) in southeastern Queensland, Australia

A population of free-ranging koalas in southeastern Queensland was examined to determine the prevalence of Chlamydia psittaci infections. Although C. psittaci was isolated from 46 of 65 (71%) koalas studied, only six (9%) of these had clinical signs of disease. Most adult females (82%) had back or pouch young present even though 67% of them were infected. There were no significant correlations between age, sex or site of sampling (urogenital versus conjunctival tissues) and the isolation of C. psittaci. No other important bacterial or fungal pathogens were isolated. The complement fixation test had a sensitivity of 7% and a specificity of 94% in detecting chlamydial infections, suggesting that it is unsuitable for use as a screening test. Chlamydia psittaci infection within this population appeared to represent a generally well-balanced host-parasite relationship and few animals had clinical signs of disease. Only four of 27 (15%) healthy koalas infected with C. psittaci followed for 24 wk after sampling developed eye disease or "dirty tail." Two koalas with keratoconjunctivitis recovered without treatment during the study period. Additional factors, including the stresses imposed by loss of habitat, may act to produce overt disease in koalas with latent C. psittaci infections.     

Arbovirus and insect‐specific virus discovery in Kenya by novel six genera multiplex high‐resolution melting analysis

A broad diversity of arthropod‐borne viruses (arboviruses) of global health concern are endemic to East Africa, yet most surveillance efforts are limited to just a few key viral pathogens. Additionally, estimates of arbovirus diversity in the tropics are likely to be underestimated as their discovery has lagged significantly over past decades due to limitations in fast and sensitive arbovirus identification methods. Here, we developed a nearly pan‐arbovirus detection assay that uses high‐resolution melting (HRM) analysis of RT–PCR products from highly multiplexed assays to differentiate broad diversities of arboviruses. We differentiated 15 viral culture controls and seven additional synthetic viral DNA sequence controls, within Flavivirus, Alphavirus, Nairovirus, Phlebovirus, Orthobunyavirus and Thogotovirus genera. Among Bunyamwera, sindbis, dengue and Thogoto virus serial dilutions, detection by multiplex RT–PCR‐HRM was comparable to the gold standard Vero cell plaque assays. We applied our low‐cost method for enhanced broad‐range pathogen surveillance from mosquito samples collected in Kenya and identified diverse insect‐specific viruses, including a new clade in anopheline mosquitoes, and Wesselsbron virus, an arbovirus that can cause viral haemorrhagic fever in humans and has not previously been isolated in Kenya, in Culex spp. and Anopheles coustani mosquitoes. Our findings demonstrate how multiplex RT–PCR‐HRM can identify novel viral diversities and potential disease threats that may not be included in pathogen detection panels of routine surveillance efforts. This approach can be adapted to other pathogens to enhance disease surveillance and pathogen discovery efforts, as well as the study of pathogen diversity and viral evolutionary ecology.     

Changes in Kit Fox Defecation Patterns During the Reproductive Season: Implications for Noninvasive Surveys

Abstract: Noninvasive survey methods based on analyzing DNA extracted from feces can be useful for carnivores that are difficult to study by other methods. Changes in fecal deposition patterns associated with reproduction in kit foxes (Vulpes macrotis) might affect results of such surveys. We used a trained dog to collect fresh scats on 2-km transects in the home ranges of 11 radiocollared female kit foxes in January, February, and March 2008 and determined sex of the individual that deposited the scats by amplifying the zinc finger protein gene. Female foxes give birth in mid-February to mid-March. We found a similar number of scats each month. In January, the sex ratio of the scats was not different from the expected 1:1. However, in February there were almost 2 male scats for every female scat and in March there were >8 male scats for every female scat. Comparing March to January, there were more male scats on all 11 transects and fewer female scats on 10 of 11 transects. Around the time pups are born, both sexes appear to show changes in fecal deposition patterns that make it easier to find male scats and harder to find female scats. Effects of these changes on survey results will vary depending on the purpose and design of the survey. Surveys to determine distribution and relative abundance would probably not be negatively affected by these changes. However, if surveys to estimate abundance are conducted during the reproductive season, they could result in an underestimate of population size unless the increased heterogeneity in scat detectability is taken into account.     

Effectiveness of Scat Detection Dogs for Detecting Forest Carnivores

Abstract: We assessed the detection and accuracy rates of detection dogs trained to locate scats from free-ranging black bears (Ursus americanus), fishers (Martes pennanti), and bobcats (Lynx rufus). During the summers of 2003-2004, 5 detection teams located 1,565 scats (747 putative black bear, 665 putative fisher, and 153 putative bobcat) at 168 survey sites throughout Vermont, USA. Of 347 scats genetically analyzed for species identification, 179 (51.6%) yielded a positive identification, 131 (37.8%) failed to yield DNA information, and 37 (10.7%) yielded DNA but provided no species confirmation. For 70 survey sites where confirmation of a putative target species' scat was not possible, we assessed the probability that ≤1 of the scats collected at the site was deposited by the target species (probability of correct identification; P_ID). Based on species confirmations or P_ID values, we detected bears at 57.1% (96) of sites, fishers at 61.3% (103) of sites, and bobcats at 12.5% (21) of sites. We estimated that the mean probability of detecting the target species (when present) during a single visit to a site was 0.86 for black bears, 0.95 for fishers, and 0.40 for bobcats. The probability of detecting black bears was largely unaffected by site- or visit-specific covariates, but the probability of detecting fishers varied by detection team. We found little or no effect of topographic ruggedness, vegetation density, or local weather (e.g., temp, humidity) on detection probability for fishers or black bears (data were insufficient for bobcat analyses). Detection dogs were highly effective at locating scats from forest carnivores and provided an efficient and accurate method for collecting detection-nondetection data on multiple species.     

Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in <Emphasis Type="Italic">Chlamydia</Emphasis>-triggered reactive arthritis

Reports of the use of HLA-B27/peptide tetrameric complexes to study peptide-specific CD8⁺ T cells in HLA-B27⁺-related diseases are rare. To establish HLA-B27 tetramers we first compared the function of HLA-B27 tetramers with HLA-A2 tetramers by using viral epitopes. HLA-B27 and HLA-A2 tetramers loaded with immunodominant peptides from Epstein–Barr virus were generated with comparable yields and both molecules detected antigen-specific CD8⁺ T cells. The application of HLA-B27 tetramers in HLA-B27-related diseases was performed with nine recently described Chlamydia-derived peptides in synovial fluid and peripheral blood, to examine the CD8⁺ T cell response against Chlamydia trachomatis antigens in nine patients with Chlamydia-triggered reactive arthritis (Ct-ReA). Four of six HLA-B27⁺ Ct-ReA patients had specific synovial T cell binding to at least one HLA-B27/Chlamydia peptide tetramer. The HLA-B27/Chlamydia peptide 195 tetramer bound to synovial T cells from three of six patients and HLA-B27/Chlamydia peptide 133 tetramer to synovial T cells from two patients. However, the frequency of these cells was low (0.02–0.09%). Moreover, we demonstrate two methods to generate HLA-B27-restricted T cell lines. First, HLA-B27 tetramers and magnetic beads were used to sort antigen-specific CD8⁺ T cells. Second, Chlamydia-infected dendritic cells were used to stimulate CD8⁺ T cells ex vivo. Highly pure CD8 T cell lines could be generated ex vivo by magnetic sorting by using HLA-B27 tetramers loaded with an EBV peptide. The frequency of Chlamydia-specific, HLA-B27 tetramer-binding CD8⁺ T cells could be increased by stimulating CD8⁺ T cells ex vivo with Chlamydia-infected dendritic cells. We conclude that HLA-B27 tetramers are a useful tool for the detection and expansion of HLA-B27-restricted CD8⁺ T cells. T cells specific for one or more of three Chlamydia-derived peptides were found at low frequency in synovial fluid from HLA-B27⁺ patients with Ct-ReA. These cells can be expanded ex vivo, suggesting that they are immunologically functional.     

Extrusions are phagocytosed and promote Chlamydia survival within macrophages

The precise strategies that intracellular pathogens use to exit host cells have a direct impact on their ability to disseminate within a host, transmit to new hosts, and engage or avoid immune responses. The obligate intracellular bacterium Chlamydia trachomatis exits the host cell by two distinct exit strategies, lysis and extrusion. The defining characteristics of extrusions, and advantages gained by Chlamydia within this unique double‐membrane structure, are not well understood. Here, we define extrusions as being largely devoid of host organelles, comprised mostly of Chlamydia elementary bodies, and containing phosphatidylserine on the outer surface of the extrusion membrane. Extrusions also served as transient, intracellular‐like niches for enhanced Chlamydia survival outside the host cell. In addition to enhanced extracellular survival, we report the key discovery that chlamydial extrusions are phagocytosed by primary bone marrow‐derived macrophages, after which they provide a protective microenvironment for Chlamydia. Extrusion‐derived Chlamydia staved off macrophage‐based killing and culminated in the release of infectious elementary bodies from the macrophage. Based on these findings, we propose a model in which C. trachomatis extrusions serve as “trojan horses” for bacteria, by exploiting macrophages as vehicles for dissemination, immune evasion, and potentially transmission.     

Mammalian Diet and Broad Hunting Strategy of the Dingo (Canis familiaris dingo) in the Wet Tropical Rain Forests of Northeastern Australia1

The diet of dingoes (Canis familiaris dingo) in the Australian Wet Tropics was examined by analyzing 383 dingo scats collected throughout the region for the presence of mammal prey remains. The scats yielded 29 native and 4 introduced mammal prey species from 14 families. The most important species in terms of percentage occurrence in the scats were Melomys cervinipes (22.2%), hoodon macrourus (17.0%), Perameles nasuta (12.5%), and Thylogale stigmatica (12.5%). The most important families were Muridae (37.1%), Peramelidae (29.5%), and Macropodidae (25.8%). Examination of small‐scale habitat preferences revealed species that preferentially use the forest edge ranked significantly higher in the diet than those that do not, and species that are terrestrial ranked higher in the diet than those that are arboreal. Relative abundance was also a significant factor in the ranked dietary occurrence of each species, with abundant species ranked significantly higher than those that are less abundant. These results suggest that dingoes in the Australian Wet Tropics are opportunistic predators of a wide variety of mammal species, with abundant terrestrial and forest edge‐dwelling taxa the most susceptible to predation.     

Genetic structure and diversity of the koala population in South Gippsland,Victoria: a remnant population of high conservation significance

In the Australian state of Victoria, the history of koalas and their management has resulted in the homogenisation and reduction of genetic diversity in many contemporary populations. Decreased genetic diversity may reduce a species’ ability to adapt to future environmental pressures such as climate change or disease. The South Gippsland koala population is considered to be unique in Victoria, as it is believed to be a remnant population, not originating from managed populations that have low genetic variation. This study investigated genetic structure and diversity of koalas in South Gippsland, with comparison to other populations in Victoria (French Island/Cape Otway, FI and Raymond Island, RI), New South Wales and south east Queensland. Population analyses were undertaken using both microsatellite genotype and mitochondrial DNA sequence data. Non-invasive sampling of koala scats was used to source koala DNA, allowing 222 South Gippsland koalas to be genotyped. Using nuclear data the South Gippsland koala population was found to be significantly differentiated (D_jost 95% CI SG–RI?=?0.03–0.06 and SG–FI?=?0.08–012) and more diverse (A_R 95% CI SG?=?4.7–5.6, RI?=?3.1–3.3, FI?=?3.0–3.3; p?=?0.001) than other Victorian koala populations, supporting the premise that koalas in the South Gippsland region are part of a remnant population, not derived from translocated island stock. These results were also supported by mitochondrial data where eight haplotypes (Pc4, Pc17, Pc26, Pc27, and Pc56–Pc59) were identified in South Gippsland while a single haplotype (Pc27) was found in all island koalas tested. Compared to other Victorian koala populations, greater genetic diversity found in South Gippsland koalas, may provide this population with a greater chance of survival in the face of future environmental pressures. The South Gippsland koala population is, therefore, of high conservation significance, warranting the implementation of strategies to conserve this population and its diversity into the future.     

Influences on koala habitat selection across four local government areas on the far north coast of NSW

Conserving habitats crucial for threatened koala (Phascolarctos cinereus) populations requires rating habitat quality from a fine spatial scale to patches, landscapes and then regions. The koala has a specialized diet focused on the leaves of a suite of Eucalyptus species. We asked: what are the key regional influences on habitat selection by koalas in the far north coast of New South Wales? We addressed this question by investigating the multi-scale factors, and within-scale and cross-scale interactions, that influence koala habitat selection and distribution across four local government areas on the far north coast of New South Wales. We assembled and analysed a large data set of tree selection, identified by the presence of scats, in a wide range of randomly selected 5 × 5 km grids across the region. This resulted in more than 9000 trees surveyed for evidence of koala use from 302 field sites, together with associated biophysical site features. The dominant factor influencing habitat use and koala occurrence was the distribution of five Eucalyptus species. Koalas were more likely to use medium-sized trees of these species where they occurred on soils with high levels of Colwell phosphorous. We also identified new interactions among the distribution of preferred tree species and soil phosphorous, and their distribution with the amount of suitable habitat in the surrounding landscape. Our study confirmed that non-preferred species of eucalypts and non-eucalypts are extensively used by koalas and form important components of koala habitat. This finding lends support to restoring a mosaic of koala-preferred tree species and other species recognized for their value as shelter. Our study has provided the ecological foundation for developing a novel regional-scale approach to the conservation of koalas, with adaptability to other wildlife species.