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Q fever, caused by Coxiella burnetii , is an important zoonotic and public health concern worldwide. Kangaroos are thought to be a likely wildlife reservoir for C. burnetii in Australia and the kangaroo tick (Amblyomma triguttatum) has often been considered a vector. In this descriptive study of ticks collected from a population of eastern grey kangaroos (Macropus giganteus) with a high serological (84 %) and molecular (65 %) prevalence of C. burnetii in northern New South Wales, a total of 72 A. triguttatum ticks were tested by PCRs targeting the IS 1111, htpAB , and com1 genes of the Coxiella genome. Despite the remarkably high prevalence of coxiellosis in the host population, none of the ticks were positive for Coxiella. This finding suggests that the kangaroo tick may not play a significant role in the transmission dynamics of C. burnetii in this particular host population. [ABSTRACT FROM AUTHOR]

Abstract 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. Graphical abstract Unlabelled Image Highlights • Exposure models can predict toxic effects of fluoride consumption in wildlife. • Exposure models that vary in intensity of field data collection warrant comparison. • Simple spatial metrics can predict fluoride accumulation in a free-ranging mammal. • Complex exposure models may not perform better than simple spatial metrics. [ABSTRACT FROM AUTHOR]

Poxviruses have previously been detected in macropods with cutaneous papillomatous lesions, however to date, no comprehensive analysis of a poxvirus from kangaroos has been performed. Here we report the genome sequences of a western grey kangaroo poxvirus (WKPV) and an eastern grey kangaroo poxvirus (EKPV), named for the host species from which they were isolated, western grey ( Macropus fuliginosus ) and eastern grey ( Macropus giganteus ) kangaroos. Poxvirus DNA from WKPV and EKPV was isolated and entire coding genome regions determined through Roche GS Junior and Illumina Miseq sequencing, respectively. Viral genomes were assembled using MIRA and SPAdes, and annotations performed using tools available from the Viral Bioinformatics Resource Centre. Histopathology and transmission electron microscopy analysis was also performed on WKPV and its associated lesions. The WKPV and EKPV genomes show 96% identity (nucleotide) to each other and phylogenetic analysis places them on a distinct branch between the established Molluscipoxvirus and Avipoxvirus genera. WKPV and EKPV are 170 kbp and 167 kbp long, containing 165 and 162 putative genes, respectively. Together, their genomes encode up to 47 novel unique hypothetical proteins, and possess virulence proteins including a major histocompatibility complex class II inhibitor, a semaphorin-like protein, a serpin, a 3-β-hydroxysteroid dehydrogenase/δ 5 → 4 isomerase, and a CD200-like protein. These viruses also encode a large putative protein (WKPV-WA-039 and EKPV-SC-038) with a C-terminal domain that is structurally similar to the C-terminal domain of a cullin, suggestive of a role in the control of host ubiquitination. The relationship of these viruses to members of the Molluscipoxvirus and Avipoxvirus genera is discussed in terms of sequence similarity, gene content and nucleotide composition. A novel genus within subfamily Chordopoxvirinae is proposed to accommodate these two poxvirus species from kangaroos; we suggest the name, Thylacopoxvirus (thylaco-: [Gr.] thylakos meaning sac or pouch). [ABSTRACT FROM AUTHOR]

Lancefield Swamp, south-eastern Australia, was one of the earliest sites to provoke interest in Pleistocene faunal extinctions in Sahul (Pleistocene Australia-New Guinea). The systematic investigation of the deposit in the early 1970s identified megafaunal remains dominated by the 100–200 kg kangaroo Macropus giganteus titan . Associated radiocarbon ages indicated that the species was extant until c.30,000 BP, suggesting significant overlap with human settlement of Sahul. This evidence was inconsistent with contemporary models of rapid human-driven extinctions. Instead, researchers inferred ecological tethering of fauna at Lancefield Swamp due to intense drought precipitated localised mass deaths, consistent with Late Pleistocene climatic variability. Later investigations in another part of the swamp, the Mayne Site, remote to the initial investigations, concluded that mass flow disturbed this area, and Electron Spin Resonance (ESR) analyses on megafauna teeth returned wide-ranging ages. To clarify site formation processes and dating of Lancefield Swamp, we excavated new test-pits next to previous trenches in the Classic and Mayne Sites. We compared absolute chronologies for sediments and teeth, sedimentology, palaeo-topography, taphonomy, and macropod age at death across the swamp. Luminescence dating of sediments and ESR analysis of teeth returned ages between c.80,000 and 45,000 years ago. We found no archaeological remains in the bone beds, and evidence of carnivore activity and fluvial action, in the form of reactivated spring flow. The latter disturbed limited parts of the site and substantial areas of the bone beds remained intact. The faunal assemblage is dominated by megafaunal adult Macropus , consistent with mass die-offs due to severe drought. Such droughts appear to have recurred over millennia during the climatic variability of Marine Isotope Stages 4 and 3. These events began tens of millennia before the first appearance of Aboriginal people in Sahul and only the very youngest fossil deposits could be coeval with the earliest human arrivals. Therefore, anthropogenic causes cannot be implicated in most if not all of mass deaths at the site. Climatic and environmental changes were the main factors in site formation and megafauna deaths at Lancefield Swamp. [ABSTRACT FROM AUTHOR]

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 M 1 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. [ABSTRACT FROM AUTHOR]

Summary Significantly elevated bone fluoride concentrations have been reported in a population of eastern grey kangaroos ( Macropus giganteus ) resident near a fluoride-emitting aluminum smelter in southeastern Australia. This paper describes the skeletal and synovial joint lesions observed post mortem in the same sample of kangaroos ( n = 76). The prevalence and severity of skeletal lesions, specifically the formation of multiple, large, smooth exostoses over the diaphysis of long bones (especially, but not exclusively, on the tibia, fibula and metatarsi), were positively associated with bone fluoride concentration. So too were lesions of degenerative joint disease, including periarticular osteophytosis, articular cartilage erosion/ulceration, synovial hyperplasia and joint capsular fibrosis. Joint lesions were most commonly seen in the knee, hock and metatarsophalangeal joints. This is the first study to describe in detail the full range of lesions induced by chronic fluorosis in a marsupial species. [ABSTRACT FROM AUTHOR]

Abstract: A total of 438 stool samples from 38 different species of animal from seven different locations were studied for the presence of Blastocystis. PCR analysis was completed on all samples and DNA sequence data from the rDNA were submitted to subtype allocation. There was a total of 80 (18%) sequences from 18 species, and nine different subtypes were identified – ST1, ST2, ST3, ST4, ST5, ST7, ST11, ST12 and ST13. This is the first report of Blastocystis from the eastern grey kangaroo, red kangaroo, wallaroo, snow leopard and ostrich. This study highlights the need for further investigation into the genetic diversity of Blastocystis which could help show the zoonotic potential of Blastocystis. [Copyright &y& Elsevier]

Abstract: Australian marsupials are unique fauna that have evolved and adapted to unique environments and thus it is likely that their detoxification systems differ considerably from those of well-studied eutherian mammals. Knowledge of these processes in marsupials is therefore vital to understanding the consequences of exposure to xenobiotics. Cytochromes P450 (CYPs) are critically important in the oxidative metabolism of a diverse array of both xenobiotics and endogenous substrates. In this study we have cloned and characterized CYP3A70, the first identified member of the CYP3A gene subfamily from Eastern gray kangaroo (Macropus giganteus). A 1665 base pair kangaroo hepatic CYP3A complete cDNA, designated CYP3A70, was cloned by reverse transcription-polymerase chain reaction approaches, which encodes a protein of 506 amino acids. The CYP3A70 cDNA shares approximately 71% nucleotide and 65% amino acid sequence homology to human CYP3A4 and displays high sequence similarity to other published mammalian CYP3As from human, monkey, cow, pig, dog, rat, rabbit, mouse, hamster, and guinea pig. Transfection of the CYP3A70 cDNAs into 293T cells resulted in stable cell lines expressing a CYP3A immuno-reactive protein that was recognized by a goat anti-human CYP3A4 polyclonal antibody. The anti-human CYP3A4 antibody also detected immunoreactive proteins in liver microsomes from all test marsupials, including the kangaroo, koala, wallaby, and wombat, with multiple CYP3A immunoreactive bands observed in kangaroo and wallaby tissues. Relatively, very low CYP catalytic activity was detected for the kangaroo CYP3A70 cDNA-expressed proteins (19.6 relative luminescent units/μg protein), which may be due to low protein expression levels. Collectively, this study provides primary molecular data regarding the Eastern kangaroo hepatic CYP3A70 gene and enables further functional analyses of CYP3A enzymes in marsupials. [Copyright &y& Elsevier]

Abstract: Toxoplasma gondii infection is frequently asymptomatic; however, it can be severe or even fatal to some hosts. In this study, diagnosis of disseminated toxoplasmosis in one red kangaroo (Macropus rufus) and one great grey kangaroo (Macropus giganteus) from the La Plata Zoo, Argentina and the isolation and molecular characterization of T. gondii are reported. Both male kangaroos showed depression and sudden death. Toxoplasma gondii infection was diagnosed by fresh examination, histophatology, immunohistochemistry, PCR and bioassay in mice. During fresh examination many protozoan cysts were observed in diaphragm, heart and hind limb muscles of M. rufus. Cysts were also observed in samples from M. giganteus, although in lower number. Cysts from both kangaroos stained strongly with T. gondii anti-serum by immunohistochemistry. The M. rufus showed more considerable histopathological lesions like non-suppurative meningoencephalitis, myositis and myocarditis. All mice inoculated with tissues from both kangaroos developed IFAT titers to T. gondii (titer ≥800) and brain cysts at necropsy. Both T. gondii isolates were maintained by mice passages and the M. rufus isolate was also maintained in cell culture. Toxoplasma gondii DNA from tissue samples was analyzed by PCR-RFLP analysis using the markers 5′SAG2, 3′SAG2, BTUB, GRA6, SAG3, c22-8, L358, PK1, c29-2 and Apico. Genotyping revealed that the T. gondii isolate from M. rufus was clonal type III and the isolate from M. giganteus was clonal type II. This is the first report of disseminated toxoplasmosis in M. rufus and M. giganteus in Argentina caused by genotypes of T. gondii considered non-virulent in a mouse model. [Copyright &y& Elsevier]

Abstract: A novel herpesvirus was detected in a captive mob of eastern grey kangaroos (Macropus giganteus) during diagnostic workup for individuals with ulcerative cloacitis. Virus was initially detected in tissues using a consensus herpesvirus PCR. No viral inclusions or particles had been evident in routine histologic or transmission electron microscopic sections of cloacal lesions. Virus was isolated from samples and transmission electron microscopy of the resulting isolates confirmed that the virus was morphologically consistent with a herpesvirus. Nucleotide sequencing of the PCR product from tissue samples and from the isolates revealed that the virus was in the subfamily Gammaherpesvirinae and was distinct from other known herpesviruses. The correlation between the lesions and the novel virus remains unknown. Two herpesviruses, both in the subfamily Alphaherpesvirinae, have previously been described in macropods and are known to cause systemic clinical disease. This is the first reported gammaherpesvirus within the order Marsupialia, and may provide valuable information regarding the evolution and phylogeny of this virus family. Based on current herpesvirus nomenclature convention, the authors propose the novel herpesvirus be named Macropodid herpesvirus 3 (MaHV-3). [Copyright &y& Elsevier]