Seroprevalence and risk factors for Toxoplasma gondii exposure in Australian feral and stray cats using an in-house modified agglutination test.

Toxoplasma gondii is a globally distributed zoonotic protist, capable of infecting all warm-blooded animals. In Australia, cats (Felis catus) are the only definitive host capable of spreading T. gondii infection via oocysts. Free-roaming cats are widespread in Australia and can play a central role in the ecology of T. gondii. Therefore, understanding the epidemiology of this parasite in stray and feral cats is essential to understanding the potential risk of infection in animals and humans. Due to a lack of easily accessible commercial kits, an in-house modified agglutination test (MAT) was established to test for IgG antibodies against T. gondii, using cell culture-derived T. gondii tachyzoites, and compared with a commercial MAT. A total of 552 serum samples collected during 2018 - 2021 from stray (n = 456) and feral cats (n = 90) (samples with missing data n = 6) from four Australian states, representing different age groups of both sexes, were screened for antibodies against T. gondii. Risk factors for T. gondii infection were assessed using multivariable logistic regression analysis. The in-house MAT had excellent agreement with the commercial MAT and provided a reliable and economical serological tool for T. gondii screening in animals. The overall observed seroprevalence for T. gondii in cats was 40.4 % (223/552). Bodyweight (as a proxy for age), geographical location, season and whether cats were feral or stray, were factors associated with T. gondii seropositivity in cats. Sex was not found to be a risk factor for T. gondii infection in feral and stray cats. This study shows that Australian stray and feral cats have a high T. gondii seroprevalence, which may translate to significant health impacts for wildlife species, livestock and the public.

Surveillance for Toxoplasma gondii, Brucella spp., and Chlamydia spp. in Australian Fur Seal (Arctocephalus pusillus doriferus) Abortions.

The intracellular pathogens Toxoplasma gondii, Brucella spp., and Chlamydia spp. are all known causative agents of abortion in wildlife. Both T. gondii and Brucella spp. have been identified in marine mammal abortions and a limited number of studies have detected their potential presence in Australian fur seals (Arctocephalus pusillus doriferus), but data are sparse for these pathogens in Australian fur seal breeding colonies. Australian fur seals have been shown to have a high degree of third-trimester pregnancy loss in one of their largest breeding colonies. Additionally, pup production has declined at the largest breeding colony for the species. This study surveyed the presence of T. gondii, Brucella spp., and Chlamydia spp. as potential infectious causes of this reproductive loss. Aborted fetuses were collected from two of the largest breeding colonies for the species, Seal Rocks (n=19) and Kanowna Island (n=34). These were examined grossly and through histopathological evaluation, in conjunction with molecular testing for all three pathogens. Placentas were collected from full-term births during the pupping season from Kanowna Island (n=118). These were used to compare the molecular prevalence of the three pathogens in presumed successful pregnancies. Chlamydia spp. was not detected in aborted fetuses in this study. Brucella spp. was detected with PCR in both aborted fetuses (9.4%) and placentas from full-term births (3.4%), and T. gondii was detected using routine histopathology (n=2/53), immunohistochemistry (n=3/4), and PCR (n=4/53) in tissues from aborted fetuses. Toxoplasma gondii was present in 7.5% of third-trimester abortions and absent from all full-term placentas. Brucella spp. was detected in both aborted fetuses and full-term placentas. This is the first description of vertical transmission of T. gondii in a marine mammal from the southern hemisphere.

Poor Survival Rate of Eastern Gray Kangaroos (Macropus giganteus) Affected by Chronic Phalaris Toxicity.

Chronic phalaris toxicity (CPT) is a neurological disease caused by animals ingesting toxins produced by early growth stages of Phalaris aquatica, a pasture plant introduced to the southeastern regions of Australia post colonization. Little is known about the clinical progression of CPT in wildlife, as incidents are sporadic and predominantly reported when animals are in the end stages of disease and in a poor welfare state. We studied a cohort of 35 eastern gray kangaroos (Macropus giganteus) affected by CPT to clarify clinical prognosis and survival rates. Kangaroos were captured in May, June, and July of 2022 at Plenty Gorge Parklands, Victoria, Australia. Each animal was radiotracked for 180 d, clinical progression and disease outcomes monitored twice a week. By the conclusion of the study, 24 animals had died (19 by euthanasia due to deterioration, five found dead). Ten animals survived, with two demonstrating a reduction in clinical signs and eight showing full resolution of clinical signs. One animal was disqualified from the study. The overall survival rate was 29.4% (95% confidence interval 17.5-49.5%). The survival duration of animals that died ranged from 5 to 133 d. There was no difference in survival rate based on sex (P=0.2), age class (P=0.49) or the month of capture (P=0.49). These results suggest that CPT is an important health and welfare concern for at-risk macropod populations, with high case-fatality rates and prolonged clinical durations. Further research to manage the disease via methods such as reducing Phalaris aquatica plant coverage and preventative treatments for animals is warranted to reduce disease incidences and improve disease outcomes in wildlife populations.

Interactions Between Heavy Metal Exposure and Blood Biochemistry in an Urban Population of the Black Swan (Cygnus atratus) in Australia.

There is growing recognition of the threat posed to wildlife by pollutants. Waterbirds are robust bioindicators of ecosystem health, and metal toxicity is a threat to these species in waterways worldwide. Urban waterbirds are likely to be at the highest risk of heavy metal exposure, but this issue has not been widely explored in Australia. Our aim was to estimate contemporary heavy metal exposure in a sedentary urban waterbird population: black swans (Cygnus atratus) inhabiting an inner-city wetland in one of Australia's largest cities, Melbourne. To investigate the physiological implications of legacy heavy metal exposure in these birds, we quantified blood biochemistry profiles and examined their relationships with metal concentrations in feathers. We caught 15 swans in 2021 and took feather samples to measure the concentration of eight heavy metals (chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), lead (Pb), and mercury (Hg)), and blood samples to measure the concentration of 13 plasma analytes. Multivariate regression analysis revealed few associations between heavy metals and biochemistry markers, and no differences between sexes or age classes. This study presents a baseline dataset of these contaminants and blood biochemical profiles of swans at this wetland that can be used for future monitoring and is an important step toward a better understanding of the threat posed by heavy metals to Australian urban waterbirds.

Exposure to Persistent Organic Pollutants in Australian Waterbirds.

There is growing worldwide recognition of the threat posed by persistent organic pollutants (POPs) to wildlife populations. We aimed to measure exposure levels to POPs in a Southern Hemisphere aquatic waterbird species, the nomadic gray teal (Anas gracilis), which is found across Australia. We collected wings from 39 ducks harvested by recreational hunters at two sites (one coastal, one inland) in Victoria, southeastern Australia, in 2021. We examined three groups of POPs: nine congeners of polychlorinated biphenyls (PCBs), 13 organochlorine pesticides (OCPs), and 12 polycyclic aromatic hydrocarbons (PAHs). The PCBs, OCPs, and PAHs were detected at quantifiable levels in 13%, 72%, and 100% of birds, respectively. Of the congeners we tested for in PCBs, OCPs, and PAHs, 33%, 38%, and 100% were detected at quantifiable levels, respectively. The highest levels of exposure to POPs that we found were to the PAH benzo[b]fluoranthene, occurring at a concentration range of 1.78 to 161.05 ng/g wet weight. There were some trends detected relating to differences between geographical sites, with higher levels of several PAHs at the coastal versus inland site. There were several strong, positive associations among PAHs found. We discuss potential sources for the POPs detected, including industrial and agricultural sources, and the likely role of large-scale forest fires in PAH levels. Our results confirm that while Australian waterbirds are exposed to a variety of POPs, exposure levels are currently relatively low. Additional future investigations are required to further characterize POPs within Australian waterbird species. Environ Toxicol Chem 2024;43:736-747. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Toxoplasma gondii Does Not Inhibit the Assisted Colonization of Eastern Barred Bandicoots (Perameles gunnii) to Phillip Island, Victoria, Australia.

Eastern barred bandicoots (Perameles gunnii) are thought to be highly susceptible to disease caused by infection with the protozoan parasite Toxoplasma gondii. This study followed a population of 67 P. gunnii introduced onto the Summerland Peninsula, Phillip Island, Australia, where the prevalence of T. gondii infection in the feral cat population was known to be very high. Prior to release, bandicoots were tested for serologic exposure to T. gondii using the modified agglutination test. A subset of bandicoots was tested on four occasions after release onto the peninsula. No seroconversion was detected at any time point. A subset of bandicoots was radiotracked after release and at two additional trapping sessions to help monitor survival. Toxoplasma gondii DNA was not detected by PCR in eight carcasses recovered for necropsy. Fourteen founder bandicoots (21% of founders) were known to be alive at 500 d post-release. A total of 29 unmarked bandicoots were trapped over the study period, confirming that the bandicoots were successfully reproducing on the island. Body weight, packed cell volume, and total plasma protein were used as measures of individual animal health; population health was inferred from these data. Body weight was significantly associated with trip number, with a general trend of increasing weight after release onto the island. This study showed that eastern barred bandicoots were able to establish a new population despite a probably high environmental load of T. gondii.

Towards a pan marsupial sero-immunological tool in the demanding field of wildlife serology: Marsupial immunoglobulin-binding capability with protein A/G, protein L and anti-kangaroo antibody.

Detection of infections in wildlife species is increasingly important to reduce the risk of spreading zoonotic and economically important parasites, understand disease epidemiology and promote the conservation of wildlife species. Serological tests are key in disease diagnosis and surveillance by detecting immunoglobulins against infectious agents. However, the need for species-specific reagents has limited the application of serological tests in wildlife species. This study evaluated the serum immunoglobulin-binding capability of polyclonal anti-kangaroo antibody and two non-species-specific reagents, including protein A/G and protein L, with the largest range of Australian marsupial species so far, including 32 species representing three major marsupial orders. Immunoglobulin-binding capability was assessed using immunoblotting, enzyme-linked immunosorbent assay and Western blot techniques. Variation in immunoglobulin-binding capability was observed between the three reagents and across the species tested, both across but also within taxonomic groups. Taxonomic distance was thus not always a good predictor of immunoglobulin-binding affinity, emphasizing the need to validate these reagents for each species separately. However, all three reagents bound with the serum immunoglobulins of most marsupial species tested. The findings of this study provide a valuable reference for species differences in affinity to protein A/G, protein L and anti-kangaroo antibody, assisting in the selection of appropriate reagents and the development of sero-immunological assays in Australian marsupials.

A Novel Marine Mammal Coxiella burnetii-Genome Sequencing Identifies a New Genotype with Potential Virulence.

The obligate intracellular bacterial pathogen Coxiella burnetii has been identified in a few species of marine mammals, some of which are showing population declines. It has been hypothesized that C. burnetii in marine mammals is a distinct genotype that varies significantly from the typical terrestrial genotypes. It appears to lack an IS1111. Isolates originating from Australian marine animals have a distinctly non-Australian profile of multiple-locus variable-number tandem-repeat analysis (MLVA). Extracted Coxiella DNA of Australian fur seal placental origin was sequenced using the Novaseq platform. Illumina 150 bp paired-end reads were filtered and trimmed with Trimgalore. The microbial community present in the sequenced genome was evaluated with Kraken and Bracken software using the NCBI database. A phylogenetic analysis was performed using 1131 core genes. Core genes were identified using Panaroo and inputted into Iqtree to determine the maximum-likelihood tree. A second phylogenetic tree was created using Rickettsiella grylii and using seven housekeeping genes. Results were compared with the C. burnetii Nine Mile RSA439 virulent genome. This new Australian marine mammal isolate of Coxiella (PG457) appears to be a novel genotype that lacks IS1111 and has a distinct MLVA signature (ms26, ms27, ms28, ms30, and ms31). The presence of genes for multiple virulence factors appears to give this genotype sufficient pathogenicity for it to be considered a possible causative agent of abortion in Australian fur seals as well as a potential zoonotic risk.

Lead exposure of mainland Australia's top avian predator.

Lead (Pb) toxicity, through ingestion of lead ammunition in carcasses, is a threat to scavenging birds worldwide, but has received little attention in Australia. We analyzed lead exposure in the wedge-tailed eagle (Aquila audax), the largest raptor species found in mainland Australia and a facultative scavenger. Eagle carcasses were collected opportunistically throughout south-eastern mainland Australia between 1996 and 2022. Lead concentrations were measured in bone samples from 62 animals via portable X-ray fluorescence (XRF). Lead was detected (concentration >1 ppm) in 84% (n = 52) of the bone samples. The mean lead concentration of birds in which lead was detected was 9.10 ppm (±SE 1.66). Bone lead concentrations were elevated (10-20 ppm) in 12.9% of samples, and severe (>20 ppm) in 4.8% of samples. These proportions are moderately higher than equivalent data for the same species from the island of Tasmania, and are comparable to data from threatened eagle species from other continents. Lead exposure at these levels is likely to have negative impacts on wedge-tailed eagles at the level of the individual and perhaps at a population level. Our results suggest that studies of lead exposure in other Australian avian scavenger species are warranted.

Epidemiology of Anaplasma marginale and Anaplasma centrale infections in African buffalo (Syncerus caffer) from Kruger National Park, South Africa.

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Unravelling the Diversity of Microorganisms in Ticks from Australian Wildlife.

Ticks and tick-borne pathogens pose a significant threat to the health and welfare of humans and animals. Our knowledge about pathogens carried by ticks of Australian wildlife is limited. This study aimed to characterise ticks and tick-borne microorganisms from a range of wildlife species across six sites in Victoria, Australia. Following morphological and molecular characterisation (targeting 16S rRNA and cytochrome c oxidase I), tick DNA extracts (n = 140) were subjected to microfluidic real-time PCR-based screening for the detection of microorganisms and Rickettsia-specific real-time qPCRs. Five species of ixodid ticks were identified, including Aponomma auruginans, Ixodes (I.) antechini, I. kohlsi, I. tasmani and I. trichosuri. Phylogenetic analyses of 16S rRNA sequences of I. tasmani revealed two subclades, indicating a potential cryptic species. The microfluidic real-time PCR detected seven different microorganisms as a single (in 13/45 ticks) or multiple infections (27/45). The most common microorganisms detected were Apicomplexa (84.4%, 38/45) followed by Rickettsia sp. (55.6%, 25/45), Theileria sp. (22.2% 10/45), Bartonella sp. (17.8%, 8/45), Coxiella-like sp. (6.7%, 3/45), Hepatozoon sp. (2.2%, 1/45), and Ehrlichia sp. (2.2%, 1/45). Phylogenetic analyses of four Rickettsia loci showed that the Rickettsia isolates detected herein potentially belonged to a novel species of Rickettsia. This study demonstrated that ticks of Australian wildlife carry a diverse array of microorganisms. Given the direct and indirect human-wildlife-livestock interactions, there is a need to adopt a One Health approach for continuous surveillance of tick-associated pathogens/microorganisms to minimise the associated threats to animal and human health.

Pacific Gulls (Larus pacificus) as Potential Vectors of Coxiella burnetii in an Australian Fur Seal Breeding Colony.

Recently, Coxiella burnetii has been described as a novel pathogen potentially contributing to decreased pup production in Australian fur seals (AusFS, Arctocephalus pusillus doriferus). Pacific gulls (PGs, Larus pacificus) are known to scavenge AusFS placental material during the fur seal breeding season. It is hypothesized that PGs may act as vectors for this pathogen. In the present study, cloacal swabs, oral swabs and serum were collected from PGs on Kanowna Island (KI, an AusFS breeding colony) and a nearby island, Seal Island (SI), not occupied by pinnipeds. All sample sets were evaluated with qPCR for the com1, htpAB and IS1111 markers. Most oral and cloacal swabs from KI tested positive on both the com1 (94.1%; 88.2%) and htpAB targets (76.5%; 76.5%). Amplification was very low from the SI oral swabs and cloacal swabs. Only the KI serum samples had amplification (17.7% for both com1 and htpAB). There was no IS1111 amplification in either colony. The results demonstrate that PGs can potentially act as vectors for the spread of C. burnetii. In some birds, C. burnetii was detectable in the serum, indicating that gulls can experience bacteraemia. It appears that different feeding strategies in the same species within the same ecosystem can have profound effects on the prevalence of pathogens. Further studies are required to better understand the epidemiology and potential risks of this organism.

Expanding the use of portable XRF to monitor lead exposure in an Australian duck species two decades after a ban on lead shot.

There is growing worldwide recognition of the threat posed by toxic lead for wildlife and humans. Lead toxicity from ammunition has been shown to be a threat to waterbirds across the globe. Lead shot was banned for all waterfowl hunting in Victoria, Australia, in 2002. However, no assessments of lead exposure in Australian waterfowl have been published since the 1990s. Our aim was to estimate contemporary lead exposure via measuring bone lead concentrations in a harvested dabbling duck, the Pacific black duck (Anas superciliosa). We collected wings from 77 Pacific black ducks, spanning 2018 (n = 30) and 2021 (n = 47), from nine sites with long-term histories of regular waterfowl hunting. We sought to validate portable X-ray fluorescence (XRF) for this purpose by taking a piece of humerus bone from each bird, and measuring lead concentration (mg/kg), first via non-destructive XRF and then via destructive inductively coupled plasma mass spectrometry (ICP-MS) and validated the relationship via regression analysis. Portable XRF bone lead measurement demonstrated a strong correlation with ICP-MS results using root-transformed regression (R2 = 0.85). Greater than 92 % of ducks had only background lead exposure (<10 mg/kg). When compared to historical studies in the same species at similar field sites from the 1990s, lead exposure levels were considerably lower, with mean lead concentrations ∼2-fold lower (3.7 c.f. 7.7 mg/kg), and the frequency of birds with severe lead exposure (>20 mg/kg) ∼3-fold lower (2.6 c.f. 7.5 %). Our results confirm that portable XRF is a useful option for measurement of bone lead in Australasian waterbird species. Our findings also demonstrate that a ban on the use of lead shot around 20 years ago has been associated with a substantial reduction in lead exposure in at least one species of waterfowl.

An assessment of ectoparasites across highland and lowland populations of Leadbeater's possum (Gymnobelideus leadbeateri): Implications for genetic rescue translocations.

Leadbeater's possum (Gymnobelideus leadbeateri) is a nocturnal arboreal marsupial with a restricted range centered on the Victorian Central Highlands, south-eastern Australia. Most populations inhabit wet montane ash forest and subalpine woodland, with one notable exception - a small, outlying and genetically-distinct lowland population inhabiting swamp forest at Yellingbo, Victoria. The species has been listed as critically endangered since 2015. Translocations are the mainstay of critical genetic rescue and this study explores the ectoparasites that are 'along for the ride' during translocation activities. Ectoparasites (133 fleas, 15 ticks and 76 mites) were collected opportunistically from 24 Leadbeater's possum colonies during population monitoring and genetic sampling across the lowland and highland populations. The composition of the flea assemblage varied by habitat type. Significantly greater numbers of the general marsupial fleas Acanthopsylla r. rothschildii. and Choristopsylla tristis (as a proportion of total flea numbers) were detected in lowland habitats, compared to highland habitats (Fishers exact test, P < 0.0001). Two host-specific flea species, Stephanocircus domrowi and Wurunjerria warnekei were detected only on possums in highland habitats. As a proportion of total fleas this was significantly different to possums in lowland habitats (Fisher's exact test, P = 0.0042 and P < 0.0001, respectively). Wurunjerria warnekei was suspected to be extinct prior to this study. Ticks (Ixodes tasmanii, n = 15) and mites (Haemdoelaps cleptus, n = 47 and H. anticlea, n = 29) have been identified in Leadbeater's possums historically. The possible causes of the different flea assemblages may be environmental/climatic, or due to the historic geographic division between highland and lowland animals. The planned translocations of highland individuals to lowland habitats will expose lowland individuals to novel species of previously exclusively highland fleas with unknown indirect consequences, thus careful monitoring will be required to manage any potential risks.

Alien Smooth Newts (Lissotriton vulgaris) in Australia Are Infected with Batrachochytrium dendrobatidis but Test Negative for Ranaviruses.

Smooth newts (Lissotriton vulgaris) established recently in Melbourne, Australia. Previously, the population's disease status was unknown. Samples from 34 adults and 78 larvae, collected 2011-16, were tested for two pathogens driving the global amphibian extinction crisis. The fungus Batrachochytrium dendrobatidis was identified (6.3% quantitative PCR positive); ranaviruses were not detected.

Descriptive Comparison of ELISAs for the Detection of Toxoplasma gondii Antibodies in Animals: A Systematic Review.

Toxoplasma gondii is the zoonotic parasite responsible for toxoplasmosis in warm-blooded vertebrates. This systematic review compares and evaluates the available knowledge on enzyme-linked immunosorbent assays (ELISAs), their components, and performance in detecting T. gondii antibodies in animals. Four databases were searched for published scientific studies on T. gondii and ELISA, and 57 articles were included. Overall, indirect (95%) and in-house (67%) ELISAs were the most used types of test among the studies examined, but the 'ID Screen® Toxoplasmosis Indirect Multi-species' was common among commercially available tests. Varying diagnostic performance (sensitivity and specificity) and Kappa agreements were observed depending on the type of sample (serum, meat juice, milk), antigen (native, recombinant, chimeric) and antibody-binding reagents used. Combinations of recombinant and chimeric antigens resulted in better performance than native or single recombinant antigens. Protein A/G appeared to be useful in detecting IgG antibodies in a wide range of animal species due to its non-species-specific binding. One study reported cross-reactivity, with Hammondia hammondi and Eimeria spp. This is the first systematic review to descriptively compare ELISAs for the detection of T. gondii antibodies across different animal species.

Comparison of the modified agglutination test and real-time PCR for detection of Toxoplasma gondii exposure in feral cats from Phillip Island, Australia, and risk factors associated with infection.

Toxoplasma gondii is considered a disease risk for many native Australian species. Feral cats are the key definitive host of T. gondii in Australia and therefore, investigating the epidemiology of T. gondii in cat populations is essential to understanding the risk posed to wildlife. Test sensitivity and specificity are poorly defined for diagnostic tests targeting T. gondii in cats and there is a need for validated techniques. This study focused on the feral cat population on Phillip Island, Victoria, Australia. We compared a novel real-time PCR (qPCR) protocol to the modified agglutination test (MAT) and used a Bayesian latent class modelling approach to assess the diagnostic parameters of each assay and estimate the true prevalence of T. gondii in feral cats. In addition, we performed multivariable logistic regression to determine risk factors associated with T. gondii infection in cats. Overall T. gondii prevalence by qPCR and MAT was 79.5% (95% confidence interval 72.6-85.0) and 91.8% (84.6-95.8), respectively. Bayesian modelling estimated the sensitivity and specificity of the MAT as 96.2% (95% credible interval 91.8-98.8) and 82.1% (64.9-93.6), and qPCR as 90.1% (83.6-95.5) and 96.0% (82.1-99.8), respectively. True prevalence of T. gondii infection in feral cats on Phillip Island was estimated as 90.3% (83.2-95.1). Multivariable logistic regression analysis indicated that T. gondii infection was positively associated with weight and this effect was modified by season. Cats trapped in winter had a high probability of infection, regardless of weight. The present study suggests qPCR applied to tissue is a highly sensitive, specific and logistically feasible tool for T. gondii testing in feral cat populations. Additionally, T. gondii infection is highly prevalent in feral cats on Phillip Island, which may have significant impacts on endemic and introduced marsupial populations.

TRAUMA FOUND TO BE A SIGNIFICANT CAUSE OF DEATH IN A PATHOLOGICAL INVESTIGATION OF BENT-WINGED BATS ( MINIOPTERUS ORIANAE ).

Determining the causes of mortality in endangered species is essential to understanding the possible reasons behind their decline and to facilitating the implementation of mitigating steps. The southern bent-winged bat (Miniopterus orianae bassanii) is a critically endangered Australian bat whose population numbers have decreased over the past 50 years. As part of a larger investigation to determine if disease could be a contributing factor to the decline, 27 southern bent-winged bats and one closely related eastern bent-winged bat (Miniopterus orianae oceanensis) that died during the study were necropsied and examined histologically. Trauma was the most common cause of death in the southern bent-winged bats, which mostly occurred at one site where fencing and other infrastructure was positioned around a key breeding cave. In response to these findings, management actions have been implemented to reduce this infrastructure-associated mortality of southern bent-winged bats. The single eastern bent-winged bat examined had a severe dermatitis caused by the mite Notoedres muris.

Caver Knowledge and Biosecurity Attitudes Towards White-Nose Syndrome and Implications for Global Spread.

White-nose syndrome (WNS), caused by the fungus Pseudogymnoascus destructans, has caused catastrophic declines of bat populations in North America. Risk assessment indicates that cavers could pose a risk for the spread of the fungus, however, information on cavers' knowledge of WNS and their caving and biosecurity habits is lacking. An anonymous qualitative survey was completed by delegates (n = 134) from 23 countries at an international speleological conference in Sydney, Australia. Cavers indicated that they visit caves frequently (80.6% at least bimonthly), including outside of their own country, but 20.3% of respondents did not know about WNS prior to the conference. Some respondents were incorrect, or unsure, about whether they had visited caves in countries where P. destructans occurs (26.5%) or whether their own country was free of the fungus (7.8%). Although 65.9% of respondents were aware of current decontamination protocols, only 23.9% and 31.2% (when in Australian or overseas caves, respectively) fully adhered to them. Overall, cavers showed strong willingness to help prevent further spread of this disease, but further efforts at education and targeted biosecurity activities may be urgently needed to prevent the spread of P. destructans to Australia and to other unaffected regions of the world.

When less is more: a comparison of models to predict fluoride accumulation in free-ranging kangaroos.

Vegetation contaminated by industrial fluoride emissions can cause disease in herbivorous mammals. Spatially explicit exposure models offer a quantitative approach for evaluating and managing the potentially toxic effects of chronic fluoride consumption on wildlife. We monitored eastern grey kangaroos (Macropus giganteus) inhabiting a high-fluoride environment in the buffer zone of an aluminium smelter in southeastern Australia between 2010 and 2013. We measured fluoride levels at 19 pasture sites and determined the foraging range of 37 individual kangaroos. A series of generalised linear models were developed to estimate bone fluoride accumulation as a function of pasture exposure. Model outputs were compared to identify the most appropriate predictive tool for kangaroo bone fluoride accumulation relative to exposure. Accounting for age there was a negative association between bone fluoride concentration and distance of the central emission point from both the mean centre of foraging range and the point of death. The mean foraging range centre was the best predictor, with point of death just as suitable (and simpler), whereas more complex parameters such as monthly and cumulative fluoride exposure were poor predictors of bone fluoride concentration. The more complex dietary fluoride exposure estimates did not improve predictive capability compared with the simple, spatial models. We conclude that in actively managed wildlife populations, simple, locally validated models can provide estimates of bone fluoride accumulation sufficient to support decision-making.