Silent killers? The widespread exposure of predatory nocturnal birds to anticoagulant rodenticides.

Anticoagulant rodenticides (ARs) influence predator populations and threaten the stability of ecosystems. Understanding the prevalence and impact of rodenticides in predators is crucial to inform conservation planning and policy. We collected dead birds of four nocturnal predatory species across differing landscapes: forests, agricultural, urban. Liver samples were analysed for eight ARs: three First Generation ARs (FGARs) and five SGARs (Second Generation ARs). We investigated interspecific differences in liver concentrations and whether landscape composition influenced this. FGARs were rarely detected, except pindone at low concentrations in powerful owls Ninox strenua. SGARs, however, were detected in every species and 92 % of birds analysed. Concentrations of SGARs were at levels where potential toxicological or lethal impacts would have occurred in 33 % of powerful owls, 68 % of tawny frogmouths Podargus strigoides, 42 % of southern boobooks N. bookbook and 80 % of barn owls Tyto javanica. When multiple SGARs were detected, the likelihood of potentially lethal concentrations of rodenticides increased. There was no association between landscape composition and SGAR exposure, or the presence of multiple SGARs, suggesting rodenticide poisoning is ubiquitous across all landscapes sampled. This widespread human-driven contamination in wildlife is a major threat to wildlife health. Given the high prevalence and concentrations of SGARs in these birds across all landscape types, we support the formal consideration of SGARs as a threatening process. Furthermore, given species that do not primarily eat rodents (tawny frogmouths, powerful owls) have comparable liver rodenticide concentrations to rodent predators (southern boobook, eastern barn owl), it appears there is broader contamination of the food-web than anticipated. We provide evidence that SGARs have the potential to pose a threat to the survival of avian predator populations. Given the functional importance of predators in ecosystems, combined with the animal welfare impacts of these chemicals, we propose governments should regulate the use of SGARs.

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.

Widespread exposure of powerful owls to second-generation anticoagulant rodenticides in Australia spans an urban to agricultural and forest landscape.

The powerful owl (Ninox strenua) is a threatened apex predator that consumes mainly arboreal marsupial prey. Low density populations reside in urban landscapes where their viability is tenuous. The catalyst for this research was the reported death of eight powerful owls around Melbourne, Australia, in less than one year (2020/2021). Eighteen deceased owls were toxicologically screened. We assessed toxic metals (Mercury Hg, Lead Pb, Cadmium Cd and Arsenic As) and anticoagulant rodenticides (ARs) in liver (n = 18 owls) and an extensive range of agricultural chemicals in muscle (n = 14). Almost all agricultural chemicals were below detection limits except for p,p-DDE, which was detected in 71% of birds at relatively low levels. Toxic metals detected in some individuals were generally at low levels. However, ARs were detected in 83.3% of powerful owls. The most common second-generation anticoagulant rodenticide (SGAR) detected was brodifacoum, which was present in every bird in which a rodenticide was detected. Brodifacoum was often present at toxic levels and in some instances at potentially lethal levels. Presence of brodifacoum was detected across the complete urban-forest/agriculture gradient, suggesting widespread exposure. Powerful owls do not scavenge but prey upon arboreal marsupials, and generally not rodents, suggesting that brodifacoum is entering the powerful owl food web via accidental or deliberate poisoning of non-target species (possums). We highlight a critical need to investigate SGARs in food webs globally, and not just in species directly targeted for poisoning or their predators.

Per- and polyfluoroalkyl substances (PFAS) in livestock and game species: A review.

Per- and polyfluoroalkyl substances (PFAS) are synthetic, organic chemicals that resist environmental breakdown. The properties that made PFAS into an industrial success also led to their persistence and bioaccumulation. As PFAS were widely used for many decades their presence is evident globally, and their persistence and potential for toxicity create concern for human, animal and environmental health. Following the precautionary principle, a reduction in human exposure is generally recommended. The most significant source of human exposure to PFAS is dietary intake (food and water) with additional exposure via dust. As PFAS concentrations have been more frequently studied in aquatic food sources, there is less understanding of exposure via terrestrial animals. To further define human exposure via animal products, it is necessary to determine PFAS concentrations and persistence in terrestrial livestock and game species. Studies assessing ambient concentrations of PFAS have noted that, aside from point sources of contamination, there is generally low input of PFAS into terrestrial agricultural food chains. However, livestock and game species may be exposed to PFAS via contaminated water, soil, substrate, air or food, and the contribution of these exposures to PFAS concentrations in food products is less well studied. This review focuses on perfluoroalkyl substances (PFAAs) and compiles information from terrestrial livestock and game species as a source of dietary exposure in humans, and discusses toxicokinetics and health effects in animals, while identifying future focus areas. Publications describing the transfer of PFAAs to farmed and hunted animals are scarce, and demonstrate large variability in distribution and elimination. We outline several relatively small, short-term studies in cattle, sheep, pigs and poultry. While negative effects have not been noted, the poultry investigations were the only studies to explicitly assess health effects. Comparative information is presented on PFAA concentrations in livestock products and edible tissues of game animals.

Management strategies for vaccinated animals after an outbreak of foot-and-mouth disease and the impact on return to trade.

An incursion of Foot-and-mouth disease (FMD) in a previously FMD-free country can cause significant economic damage from immediate and prolonged closure of FMD-sensitive markets. Whilst emergency vaccination may help contain disease, the presence of vaccinated animals complicates post-outbreak management and the recovery of FMD-free status for return to trade. We present enhancements to the Australian Animal DISease (AADIS) model that allow comparisons of post-outbreak management strategies for vaccinated animals, for the purposes of securing the earliest possible return to trade. Two case studies are provided that compare the retention of vaccinated animals with removal for waste/salvage, and the impact on recovery of FMD-sensitive markets per OIE guidelines. It was found that a vaccinate-and-retain strategy was associated with lower post-outbreak management costs, however this advantage was outweighed by significantly higher trade losses. Under the assumptions of the study there was no cost advantage to salvaging the removed vaccinated animals.

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.

Chronic excess fluoride uptake contributes to degenerative joint disease (DJD): Evidence from six marsupial species.

One of the manifestations of chronic fluoride toxicosis in mammals is skeletal fluorosis, which can include lesions of degenerative joint disease (DJD). Although DJD lesions have been less commonly studied than bone or dental lesions in relation to the pathology and epidemiology of fluoride toxicosis, there have been multiple independent studies in various species that have concluded that there appears to be an effect. The mechanisms by which fluoride affects the joints are not clear, but our data provide evidence that chronic excess dietary fluoride intake contributes to DJD. Our study is the first to specifically address the association between fluoride exposure and DJD in multiple species of free-ranging mammals. We describe levels of DJD in six marsupial species (Macropus giganteus, Notamacropus rufogriseus, Wallabia bicolor, Phascolarctos cinereus, Trichosurus vulpecula and Pseudocheirus peregrinus) inhabiting high and low fluoride environments. Lesions occurred to varying extents in all species, and lesion distribution varied with biomechanical differences in gait. In addition, we show an association (independent of age) between increasing bone fluoride concentration (as a measure of fluoride exposure) and increasing prevalence of moderate and severe DJD in five species of marsupial, which we propose does not persist at the highest levels of fluoride exposure due to selective survival bias.

Skeletal fluorosis in marsupials: A comparison of bone lesions in six species from an Australian industrial site.

In this study we explored the prevalence, type, location and severity of skeletal lesions in six species of Australian marsupial (Macropus giganteus, Notamacropus rufogriseus, Wallabia bicolor, Phascolarctos cinereus, Trichosurus vulpecula and Pseudocheirus peregrinus) from high and low-fluoride environments. Lesions occurred to varying extents in all species, and lesion distribution varied with biomechanical differences in gait and mastication. Bone fluoride levels increased with severity of periosteal hyperostosis. The mean bone fluoride concentration of individuals lacking hyperostosis (across all species, from both high and low-fluoride environments) was 1100±260µgF-/g dry bone, compared to 4300±1200µgF-/g and 6300±1200µgF-/g in those with mild and severe grade hyperostosis, respectively. Multivariable modelling showed that the probability of observing a lesion varied across species, anatomical location, age and bone fluoride concentration (in a non-linear manner). The pathological changes reported in the marsupials are consistent with the range of fluoride-related lesions described in other mammals, and biomechanical differences among the studied marsupial species offer some explanation for the degree of interspecific variability in prevalence, type, anatomical location, and severity of the lesions.

Developmental and Post-Eruptive Defects in Molar Enamel of Free-Ranging Eastern Grey Kangaroos (Macropus giganteus) Exposed to High Environmental Levels of Fluoride.

Dental fluorosis has recently been diagnosed in wild marsupials inhabiting a high-fluoride area in Victoria, Australia. Information on the histopathology of fluorotic marsupial enamel has thus far not been available. This study analyzed the developmental and post-eruptive defects in fluorotic molar enamel of eastern grey kangaroos (Macropus giganteus) from the same high-fluoride area using light microscopy and backscattered electron imaging in the scanning electron microscope. The fluorotic enamel exhibited a brownish to blackish discolouration due to post-eruptive infiltration of stains from the oral cavity and was less resistant to wear than normally mineralized enamel of kangaroos from low-fluoride areas. Developmental defects of enamel included enamel hypoplasia and a pronounced hypomineralization of the outer (sub-surface) enamel underneath a thin rim of well-mineralized surface enamel. While the hypoplastic defects denote a disturbance of ameloblast function during the secretory stage of amelogenesis, the hypomineralization is attributed to an impairment of enamel maturation. In addition to hypoplastic defects, the fluorotic molars also exhibited numerous post-eruptive enamel defects due to the flaking-off of portions of the outer, hypomineralized enamel layer during mastication. The macroscopic and histopathological lesions in fluorotic enamel of M. giganteus match those previously described for placental mammals. It is therefore concluded that there exist no principal differences in the pathogenic mechanisms of dental fluorosis between marsupial and placental mammals. The regular occurrence of hypomineralized, opaque outer enamel in the teeth of M. giganteus and other macropodids must be considered in the differential diagnosis of dental fluorosis in these species.

Interspecific variation in the diets of herbivores in an industrial environment: implications for exposure to fluoride emissions.

Atmospheric fluorides (gaseous and particulate) are deposited on, and absorbed by, vegetation. Ingested fluoride accumulates in calcified tissues of vertebrates, and if it is excessive, it may lead to dental and skeletal fluorosis. The prevalence, form and severity of the effects vary greatly between species. Foraging strategy can be an important determinant of fluoride exposure in herbivores, because foliar fluoride concentrations vary between plant species, for example, according to vertical and lateral position in the vegetation. We combined microhistological analysis of diet and analysis of foliar fluoride levels to examine interspecific variation in dietary fluoride exposure of macropodid marsupials (swamp wallaby Wallabia bicolor, red-necked wallaby Notamacropus rufogriseus and eastern grey kangaroo Macropus giganteus), in the buffer zone of an aluminium smelter in Victoria, Australia. Dietary niche differentiation between species was evident. The swamp wallaby and the red-necked wallaby were browsers or mixed feeders, depending on the classification system used. The eastern grey kangaroo was a grazer, consuming almost entirely grasses. However, foliar fluoride did not vary significantly between the main plant groups consumed. Our results indicate that interspecific variation in diet at this site is unlikely to explain variation in fluoride exposure.

Reconstructing temporal variation of fluoride uptake in eastern grey kangaroos (Macropus giganteus) from a high-fluoride area by analysis of fluoride distribution in dentine.

Trace element profiling in the incrementally formed dentine of mammalian teeth can be applied to reconstruct temporal variation of incorporation of these elements into the tissue. Using an electron microprobe, this study analysed fluoride distribution in dentine of first and third mandibular molars of free-ranging eastern grey kangaroos inhabiting a high-fluoride area, to assess temporal variation in fluoride uptake of the animals. Fluoride content in the early-formed dentine of first molars was significantly lower than in the late-formed dentine of these teeth, and was also lower than in both, the early and the late-formed dentine of third molars. As early dentine formation in M1 takes place prior to weaning, this finding indicates a lower dentinal fluoride uptake during the pre-weaning compared to the post-weaning period. This is hypothetically attributed to the action of a partial barrier to fluoride transfer from blood to milk in lactating females and a low bioavailability of fluoride ingested together with milk. Another factor contributing to lower plasma fluoride levels in juveniles compared to adults is the rapid clearance of fluoride from blood plasma in the former due to their intense skeletal growth. The combined action of these mechanisms is considered to explain why in kangaroos from high-fluoride areas, the (early-formed) first molars are not affected by dental fluorosis while the (later-formed) third and fourth molars regularly exhibit marked to severe fluorotic lesions.

Dental fluorosis and skeletal fluoride content as biomarkers of excess fluoride exposure in marsupials.

Particulate and gaseous fluoride emissions contaminate vegetation near fluoride-emitting industries, potentially impacting herbivorous wildlife in neighboring areas. Dental fluorosis has been associated with consumption of fluoride-contaminated foliage by juvenile livestock and wildlife in Europe and North America. For the first time, we explored the epidemiology and comparative pathology of dental fluorosis in Australian marsupials residing near an aluminium smelter. Six species (Macropus giganteus, Macropus rufogriseus, Wallabia bicolor, Phascolarctos cinereus, Trichosurus vulpecula, Pseudocheirus peregrinus) demonstrated significantly higher bone fluoride levels in the high (n=161 individuals), compared to the low (n=67 individuals), fluoride areas (p<0.001). Necropsy examinations of all six species from the high-fluoride area near the smelter revealed dental lesions considered characteristic of dental fluorosis in eutherian mammals. Within the high-fluoride area, 67% of individuals across the six species showed dental enamel lesions, compared to 3% in the low-fluoride areas. Molars that erupted before weaning were significantly less likely to display pathological lesions than those developing later, and molars in the posterior portion of the dental arcade were more severely fluorotic than anterior molars in all six species. The severity of dental lesions was positively associated with increasing bone fluoride levels in all species, revealing a potential biomarker of excess fluoride exposure.

Pharmacokinetics of moxidectin in the southern hairy-nosed wombat (Lasiorhinus latifrons).

Sarcoptic mange, caused by Sarcoptes scabiei var. wombati, could be a significant threat to populations of southern hairy-nosed wombats (Lasiorhinus latifrons; SHNW) in Australia. Treatment is currently based on the off-label use of various parasiticidal drugs, with limited clinical efficacy trials. Our primary aim was to determine the pharmacokinetic parameters of a macrocyclic lactone, moxidectin, to assist in the development of effective treatment protocols. Pharmacokinetic parameters were determined in four female SHNW following a single subcutaneous injection of 0.2 mg/kg moxidectin. Blood samples were collected for 38 days following injection (August-September 2008), for analysis using liquid chromatography and tandem mass spectrometry. The mean peak plasma concentration occurred at 13.6 hr, with a mean peak plasma level of 98.6 ng/ml. The mean elimination half-life was 5.03 days, resulting in a mean area under the curve of 377 ng.day/ml. The peak plasma moxidectin concentration was higher than that seen in livestock species but the plasma elimination half-life was shorter. This study suggests that a single injection of 0.2 mg/kg moxidectin may not be sufficient to clear a mange infection in this species.