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Kangaroo-related motor vehicle collisions are common but there is limited literature on this topic. Drivers confronted by kangaroos may choose to swerve or to directly collide with the kangaroo. The effect of these differing crash mechanisms, along with the effect of vehicle type or time of day, has not yet been reported.A retrospective cohort study was performed, examining patients admitted to our tertiary trauma centre for kangaroo-related motor vehicle collisions between 2000 and 2020. Data on patient demographics, crash characteristics, and hospital stay were collected and analysed.A total of 366 patients were included and were predominantly male (76%) with an median age of 40. Swerve crashes were more common (59%) than direct impact and swerving was a statistically significant predictor of reduced injury severity score on multivariable analysis (other significant factors were female sex and no rollover). Motor vehicle crashes and motorbike crashes had differing crash characteristics. Motor vehicle crashes were more likely the result of swerving, and swerving was less likely to cause ejection or require extrication but more likely to cause rollover. Motorbike crashes however, were more likely the result of head on collision and riders were more likely to be ejected from the vehicle, require extrication, or be involved in a rollover. In terms of time of day, there were more crashes at dawn and there was a trend towards higher injury severity score and length of stay for night-time crashes.For kangaroo-related motor vehicle crashes, predictors of increased injury severity score on multivariable analysis were male sex, direct impact, and rollover. Motorbikes and motor vehicles had differing crash mechanisms and characteristics, as did night-time crashes when compared to daytime or twilight crashes.IV, prognostic.

An albino infant wallaby was born to a mother with the wild-type body color. PCR and sequencing analyses of TYR (encoding tyrosinase, which is essential for melanin biosynthesis) of this albino wallaby revealed a 7.1-kb-long DNA fragment inserted in the first exon. Because the fragment carried long terminal repeats, we assumed it to be a copy of an endogenous retrovirus, which we named walb. We cloned other walb copies residing in the genomes of this species and another wallaby species. The copies exhibited length variation, and the longest copy (>8.0 kb) contained open reading frames whose deduced amino acid sequences were well aligned with those of gag, pol, and env of retroviruses. It is not known through which of the following likely processes the walb copy was inserted into TYR: endogenization (infection of a germline cell by an exogenous virus), reinfection (infection by a virus produced from a previously endogenized provirus), or retrotransposition (intracellular relocation of a provirus). In any case, the insertion into TYR is considered to have been a recent event on an evolutionary timescale because albino mutant alleles generally do not persist for long because of their deleterious effects in wild circumstances., 白いワラビーの遺伝的原因を初めて解明 --世界各地の動物園で相次いで誕生--. 京都大学プレスリリース. 2022-01-18.

The opossum (Monodelphis domestica), with its sequenced genome, ease of laboratory care and experimental manipulation, and unique biology, is the most used laboratory marsupial. Using the mammalian methylation array, we generated DNA methylation data from n = 100 opossum samples from the ear, liver, and tail. We contrasted postnatal development and later aging effects in the opossum methylome with those in mouse (Mus musculus, C57BL/6 J strain) and other marsupial species such as Tasmanian devil, kangaroos, and wallabies. While the opossum methylome is similar to that of mouse during postnatal development, it is distinct from that shared by other mammals when it comes to the age-related gain of methylation at target sites of polycomb repressive complex 2. Our immunohistochemical staining results provide additional support for the hypothesis that PRC2 activity increases with later aging in mouse tissues but remains constant in opossum tissues. We present several epigenetic clocks for opossums that are distinguished by their compatibility with tissue type (pan-tissue and blood clock) and species (opossum and human). Two dual-species human-opossum pan-tissue clocks accurately measure chronological age and relative age, respectively. The human-opossum epigenetic clocks are expected to provide a significant boost to the attractiveness of opossum as a biological model. Additional epigenetic clocks for Tasmanian devil, red kangaroos and other species of the genus Macropus may aid species conservation efforts.

The diversity of the diprotodontids provides an excellent opportunity to study how a basic marsupial cortical plan has been modified for the needs of the mammals living in different habitats. Very little is known about the connections of the cerebral cortex with the deep brain structures (basal ganglia and thalamus) in this evolutionarily significant group of mammals. In this study, we performed mapping of brain regions and connections in a diprotodontid marsupial from data obtained from an excised brain scanned in high-field (9.4 T) microstructural magnetic resonance imaging (MRI) instrument. The analysis was based on two MRI methodologies. First, high-resolution structural scans were used to map MRI visible brain regions from T1w and T2w images. Second, extensive diffusion tensor imaging (DTI) data were obtained to elucidate connectivity between brain areas using deterministic diffusion tracking of neuronal brain fibers. From the data, we were able to identify corticostriate connections between the frontal association and dorsomedial isocortex and the head of the caudate, and between the lateral somatosensory cortex and the putamen. We were also able to follow the olfactory and limbic connections by tracing fibers in the fornix, cingulum, intrabulbar part of the anterior commissure, and lateral olfactory tract. There was segregation of fibers in the anterior commissure such that olfactory connections passed through the rostroventral part and successively more dorsal cortical areas connected through more dorsal parts of the commissure. Our findings confirm a common pattern of cortical connectivity in therian mammals, even where brain expansion has occurred independently in diverse groups.

Funder: McKenzie Fellowship Program, Funder: University of Melbourne, Lead-based ammunition (gunshot and bullets) frequently leaves small lead fragments embedded in the meat of wild-shot game animals. Australia produces several commercial game meat products from wild animals harvested with lead-based ammunition and has a growing population of recreational hunters. However, no studies have previously investigated the frequency of lead fragments or lead concentrations in Australian game meat. We examined 133 Australian minced game meat items of four types for evidence of lead contamination. Samples were meat from kangaroos (Macropus and Osphranter spp.; n=36) and Bennett's wallabies (Notamacropus rufogriseus; n=28) sold for human consumption, and deer ('venison'; multiple spp.; n=32) and stubble quail (Coturnix pectoralis; n=37) harvested for private consumption by recreational hunters. All packages were studied by digital radiography to detect the presence of radio-dense fragments, assumed to be lead fragments from ammunition. Visible fragments were absent in commercially available kangaroo products, but were present in 4%, 28% and 35% of wallaby, venison and quail, respectively. Mean meat lead concentrations (mg/kg wet weight) were 0.01 ± 0.01 for kangaroo, 0.02 ± 0.01 for wallaby, 0.12 ± 0.07 for venison, and 1.76 ± 3.76 for quail. The Australian food standards threshold for livestock meat (0.1 mg/kg w.w.) was not exceeded by any kangaroo or wallaby products but was exceeded by 53% and 86% of venison and quail, respectively. Radiography only detected 35% of samples that were above the food safety threshold. While average lead concentrations in commercially available macropod (kangaroo and wallaby) meat were low, those in recreationally harvested game meat may pose health risks for hunters and associated consumers.

The extinct sthenurine (giant, short-faced) kangaroos have been proposed to have a different type of locomotor behavior to extant (macropodine) kangaroos, based both on physical limitations (the size of many exceeds the proposed limit for hopping) and anatomical features (features of the hind limb anatomy suggestive of weight-bearing on one leg at a time). Here, we use micro computerised tomography (micro-CT) scans of the pedal bones of six kangaroos, three sthenurine, and three macropodine, ranging from ~50 to 150 kg, to investigate possible differences in bone resistances to bending and cortical bone distribution that might relate to differences in locomotion. Using second moment of area analysis, we show differences in resistance to bending between the two subfamilies. Distribution of cortical bone shows that sthenurines had less resistant calcaneal tubers, implying a different foot posture during locomotion, and the long foot bones were more resistant to the medial bending stresses. These differences were the most pronounced between Pleistocene monodactyl sthenurines (Sthenurus stirlingi and Procoptodon browneorum) and the two species of Macropus (the extant M. giganteus and the extinct M. cf. M. titan) and support the hypothesis that these derived sthenurines employed bipedal striding. The Miocene sthenurine Hadronomas retains some more macropodine-like features of bone resistance to bending, perhaps reflecting its retention of the fifth pedal digit. The Pleistocene macropodine Protemnodon has a number of unique features, possibly indicative of a type of locomotion unlike the other kangaroos.

Background The subfamily Phascolostrongylinae (Superfamily Strongyloidea) comprises nematodes that are parasitic in the gastrointestinal tracts of macropodid (Family Macropodidae) and vombatid (Family Vombatidae) marsupials. Currently, nine genera and 20 species have been attributed to the subfamily Phascolostrongylinae. Previous studies using sequence data sets for the internal transcribed spacers (ITS) of nuclear ribosomal DNA showed conflicting topologies between the Phascolostrongylinae and related subfamilies. Therefore, the aim of this study was to validate the phylogenetic relationships within the Phascolostrongylinae and its relationship with the families Chabertiidae and Strongylidae using mitochondrial amino acid sequences. Methods The sequences of all 12 mitochondrial protein-coding genes were obtained by next-generation sequencing of individual adult nematodes (n = 8) representing members of the Phascolostrongylinae. These sequences were conceptually translated and the phylogenetic relationships within the Phascolostrongylinae and its relationship with the families Chabertiidae and Strongylidae were inferred from aligned, concatenated amino acid sequence data sets. Results Within the Phascolostrongylinae, the wombat-specific genera grouped separately from the genera occurring in macropods. Two of the phascolostrongyline tribes were monophyletic, including Phascolostrongylinea and Hypodontinea, whereas the tribe Macropostrongyloidinea was paraphyletic. The tribe Phascolostrongylinea occurring in wombats was closely related to Oesophagostomum spp., also from the family Chabertiidae, which formed a sister relationship with the Phascolostrongylinae. Conclusion The current phylogenetic relationship within the subfamily Phascolostrongylinae supports findings from a previous study based on ITS sequence data. This study contributes also to the understanding of the phylogenetic position of the subfamily Phascolostrongylinae within the Chabertiidae. Future studies investigating the relationships between the Phascolostrongylinae and Cloacininae from macropodid marsupials may advance our knowledge of the phylogeny of strongyloid nematodes in marsupials. Graphical Abstract

Macropods are commonly kept and exhibited species worldwide. Mortality records were reviewed for five species of macropods housed at Cleveland Metroparks Zoo from 1995 to 2016. One hundred sixty-seven deaths occurred including Western grey kangaroo (Macropus fuliginosus) (n = 22), Bennett's wallaby (M. rufogriseus) (n = 64), red kangaroo (M. rufus) (n = 56), wallaroo (M. robustus) (n = 9), and Tammar wallaby (M. eugenii) (n = 16). The leading cause of death was trauma, followed by bronchopneumonia for all species. Gastric dilatation was identified as a concern for red kangaroo. Females lived longer than males across species and were overrepresented by the data set. Oral necrobacillosis and osteomyelitis were health concerns for macropods in this review. Mortality reviews are important as they identifying collection trends and influencing long-term care.

Angiostrongylus cantonensis is a zoonotic parasitic helminth that normally resides in the pulmonary arteries and the right ventricle of rats ( Rattus sp.), the definitive host, where it causes little disease. Humans, dogs, opossums, and various zoo animals are “accidental” hosts. Here we report verminous meningoencephalomyelitis caused by A. cantonensis in a 9-mo-old male red kangaroo ( Macropus rufus). The kangaroo was first presented lethargic, recumbent, and hypothermic, with severe muscle wasting. Within 3 wk, he progressed to non-ambulatory paraparesis and died. Gross examination revealed multifocal areas of dark-brown discoloration, malacia, and cavitation in the brain and the spinal cord. Histologically, there were several sections of nematodes surrounded by extensive areas of rarefaction, hemorrhage, spongiosis, neuronal necrosis, and gliosis. Based on size, morphology, and organ location, the nematodes were identified as subadult males and females. Interestingly, an eosinophilic response was largely absent, and the inflammatory response was minimal. A. cantonensis infection had not been reported previously in a red kangaroo in Louisiana or Mississippi, to our knowledge. Our case reaffirms the widespread presence of the helminth in the southeastern United States and indicates that A. cantonensis should be considered as a differential in macropods with neurologic clinical signs in regions where A. cantonensis is now endemic.

In this study, Babesia screening was conducted in 55 rodents and 160 tick samples collected from primary forests and an oil palm plantation in Sarawak, Malaysian Borneo. PCR targeting the 18S ribosomal DNA revealed the presence of Babesia spp. DNA detected in two questing male Haemaphysalis shimoga ticks collected from the oil palm plantation. Sequence analysis revealed that both sequences were identical and had 98.6% identity to a Babesia macropus sequence obtained from Eastern grey kangaroos (Macropus giganteus) in Australia. Phylogenetic tree revealed clustering with marsupial-associated Babesia spp. in the Babesia sensu stricto clade. Whether or not H. shimoga is the competent vector and the importance of the Babesia sp. detected in this study warrants more investigation.

A privately owned 14-month-old intact female red kangaroo (Macropus rufus) was presented for acute onset respiratory distress and lethargy. On presentation, the kangaroo was laterally recumbent, tachypneic, dyspneic, lethargic, and obtunded. Thoracic radiographs revealed a severe diffuse mixed pulmonary pattern (alveolar pattern superimposed on a bronchial pattern) and subjective mild generalized cardiomegaly. Due to the severity of clinical signs and grave prognosis, euthanasia was elected. Postmortem examination was consistent with systemic toxoplasmosis. Histopathology and immunohistochemistry staining on infected tissues confirmed Toxoplasma gondii. This is the first published report of radiographic findings for confirmed toxoplasmosis in a red kangaroo or marsupial.

A molecular survey of herpesviruses in Australian native mammals was conducted, spanning 260 individuals from 27 species. Among the herpesviruses detected, a putative new gammaherpesvirus species was detected in the yellow-bellied glider (Petaurus australis), and another in the critically endangered Leadbeater's possum (Gymnobelideus leadbeateri). In addition, the known host range of the putative species macropodid gammaherpesvirus 3 (MaHV-3) is herein extended to the western grey kangaroo (Macropus fuliginosus). These findings expand our understanding of herpesviruses in Australian mammals and may inform biosecurity protocols for captive and translocated populations.

BACKGROUND Reference intervals for haematology and serum biochemistry parameters were developed for free-ranging Lumholtz's tree-kangaroo (Dendrolagus lumholtzi) using 35 samples from 12 female and 15 male free-ranging animals. Captive tree-kangaroos (n = 12) were also sampled for comparison. Differences were found between free-ranging and captive animals in white blood cell and neutrophil counts, and levels of aspartate aminotransferase, alkaline phosphatase, bilirubin, creatine kinase, phosphate, triglycerides and lipase. These differences may be attributed to diet, activity, capture methods or age group. Reference intervals generated may be used for both free-ranging and captive Lumholtz's tree-kangaroos. This study provides a valuable tool for the assessment of health in rescued and captive tree-kangaroos and will aid in investigations into population health and disease in free-ranging Lumholtz's tree-kangaroos. OBJECTIVE To develop reference intervals (RIs) for haematology and serum biochemistry parameters in Lumholtz's tree-kangaroos. METHODS Haematological and serum biochemical RIs were determined using 35 samples from 27 clinically healthy Lumholtz's tree-kangaroos from the Atherton Tablelands region of Queensland examined between 2014 and 2019. Haematology and serum biochemistry parameters were measured from 16 samples from 12 captive animals for comparison. RESULTS Reference intervals based on 35 samples from free-ranging animals showed higher mean and standard deviation values for white blood cell and neutrophil counts, and levels of aspartate aminotransferase, alkaline phosphatase, bilirubin, creatine kinase, phosphate, triglycerides and lipase than results for 16 samples from captive animals. Captive individuals showed higher mean values than free-ranging individuals for albumin, protein, creatinine as well as Hb, MCV, MCH and MCHC. CONCLUSION The haematological and serum biochemistry RIs developed for Lumholtz's tree-kangaroos in this study will provide a valuable tool during clinical examination and investigations into disease and population health by veterinarians and researchers. The differences in parameters between free-ranging and captive animals are consistent with differences in diet, age cohort, activity or capture methods. Reference intervals generated from free-ranging animals should also be valid for captive Lumholtz's tree-kangaroos.

Male germ cells undergo two consecutive processes – pre-spermatogenesis and spermatogenesis – to generate mature sperm. In eutherian mammals, epigenetic information such as DNA methylation is dynamically reprogrammed during pre-spermatogenesis, before and during mitotic arrest. In mice, by the time germ cells resume mitosis, the majority of DNA methylation is reprogrammed. The tammar wallaby has a similar pattern of germ cell global DNA methylation reprogramming to that of the mouse during early pre-spermatogenesis. However, early male germline development in the tammar or in any marsupial has not been described previously, so it is unknown whether this is a general feature regulating male germline development or a more recent phenomenon in mammalian evolutionary history. To answer this, we examined germ cell nuclear morphology and mitotic arrest during male germline development in the tammar wallaby (Macropus eugenii), a marsupial that diverged from mice and humans around 160 million years ago. Tammar pro-spermatogonia proliferated after birth and entered mitotic arrest after day 30 postpartum (pp). At this time, they began moving towards the periphery of the testis cords and their nuclear size increased. Germ cells increased in number after day 100 pp which is the time that DNA methylation is known to be re-established in the tammar. This is similar to the pattern observed in the mouse, suggesting that resumption of germ cell mitosis and the timing of DNA methylation reprogramming are correlated and conserved across mammals and over long evolutionary timescales.

As an animal grows, the relative sizes of their organs may grow proportionately or disproportionately, depending on ontogenetic changes in function. If organ growth is proportional (isometric), then the exponent of the scaling equation is 1.0. Relative decreases or increases in size result in exponents less than 1 (hypoallometric) or greater than 1 (hyperallometric). Thus, the empirical exponent can indicate potential changes in function. The entire gastrointestinal tract (GIT) of the foregut-fermenting western grey kangaroo Macropus fuliginosus melanops exhibited biphasic allometry across five orders of magnitude body mass (Mb; 52.0 g–70.5 kg). Prior to weaning at around 12 kg Mb, the entire empty GIT mass scaled with hyperallometry (Mb1.13), shifting to hypoallometry (Mb0.80) post-weaning. In addition, there were varying patterns of hyper-, hypo-, and isometric scaling for select GIT organs, with several displaying phase shifts associated with major life-history events, specifically around exit from the maternal pouch and around weaning. Mass of the kangaroo forestomach, the main fermentation site, scaled with hyperallometry (Mb1.16) before the stage of increased vegetation intake, and possibly after this stage (Mb1.12; P = 0.07), accompanied by a higher scaling factor (elevation of the curve) probably associated with more muscle for processing fibrous vegetation. The acid hindstomach mass showed hyperallometry (Mb1.15) before weaning, but hypoallometry (Mb 0.58) post-weaning, presumably associated with decreasing intake of milk. Small intestine mass and length each scaled isometrically throughout ontogeny, with no discernible breakpoints at any life-history stage. The caecum and colon mass were steeply hyperallometric early in-pouch life (Mb1.59–1.66), when the young were ectothermic, hairless, and supported solely by milk. After around 295 g Mb, caecum mass remained hyperallometric (Mb1.14), possibly supporting its early development as a nidus for microbial populations to provide for secondary fermentation in this organ after the young transition from milk to vegetation.

Explanations for the Upper Pleistocene extinction of megafauna from Sahul (Australia and New Guinea) remain unresolved. Extinction hypotheses have advanced climate or human-driven scenarios, in spite of over three quarters of Sahul lacking reliable biogeographic or chronologic data. Here we present new megafauna from north-eastern Australia that suffered extinction sometime after 40,100 (±1700) years ago. Megafauna fossils preserved alongside leaves, seeds, pollen and insects, indicate a sclerophyllous forest with heathy understorey that was home to aquatic and terrestrial carnivorous reptiles and megaherbivores, including the world’s largest kangaroo. Megafauna species diversity is greater compared to southern sites of similar age, which is contrary to expectations if extinctions followed proposed migration routes for people across Sahul. Our results do not support rapid or synchronous human-mediated continental-wide extinction, or the proposed timing of peak extinction events. Instead, megafauna extinctions coincide with regionally staggered spatio-temporal deterioration in hydroclimate coupled with sustained environmental change., The causes of the Upper Pleistocene megafauna extinction in Australia and New Guinea are debated, but fossil data are lacking for much of this region. Here, Hocknull and colleagues report a new, diverse megafauna assemblage from north-eastern Australia that persisted until ~40,000 years ago.

The discovery of antibodies against Coxiella burnetii in cattery‐confined breeding cats indicating prior or current exposure (Shapiro et al. , 2015) prompted an investigation into possible sources of infection. One hypothesis was that raw meat diets containing reservoir species may provide a source of C. burnetii transmission. The aim of this pilot study was to determine whether C. burnetii DNA was present in raw meat sold exclusively for companion animal consumption. The sample population consisted of raw meat packages (n = 58) of primarily kangaroo origin, with three to four aliquots (50–120 mg) randomly selected from each package. Genomic DNA was extracted from whole tissue in each of these aliquots using a modified protocol. Three quantitative PCR assays were used for the detection of C. burnetii targeting the IS1111 gene, the heat shock operon htpAB and the C. burnetii outer membrane protein‐coding gene, com1 . Coxiella burnetii DNA was detected in 25/58 samples (43%) using the IS1111 , htpAB and/or com1 PCR assays and confirmed by DNA sequencing. All samples amplifying a product in the com1 assay also amplified a product in the htpAB and IS1111 assays. A total of 17/58 (29%) packets were positive with all three genes, 4/58 (7%) were positive with two genes (IS1111 and htpAB ) and 4/58 (7%) were positive with the IS1111 gene only. Coxiella burnetii DNA was five times more likely to be found in offal than skeletal muscle meat samples. All meat samples in which C. burnetii DNA was found were from kangaroo tissues, while samples labelled as non‐kangaroo meat (n = 4) were negative. Multi‐locus variable number of tandem repeat analysis (MLVA) identified three different genotypes of C. burnetii that have all been identified previously from Australian human clinical Q fever cases. Further investigations are required to determine the potential role of certain raw meats in the transmission of C. burnetii to cats and humans.

Life history theory predicts trade-offs in allocation between survival, maintenance, growth, and reproduction, especially when resources are scarce. Individual variation in resource acquisition can affect trade-offs, but is often unaccounted for. We quantified the fitness costs of reproduction, accounting for environmental conditions, maternal characteristics and individual variation. We analyzed 10 years of data from marked kangaroos to evaluate how reproductive allocation affected annual mass change and skeletal growth, subsequent fecundity and weaning success, and survival, accounting for maternal mass or size and forage availability. Through repeated measurements of 76–91 females, we investigated how trade-offs varied within and between individuals, assessing whether individual variation could mask population-level trade-offs. In poor environments, females that weaned an offspring lost mass. Females that nursed an offspring for > 7 months had reduced skeletal growth. Females that did not gain mass over the previous 12 months rarely reproduced, especially if they had nursed an offspring for > 7 months the previous year. Reproductive allocation had no effect on weaning success, which was very low, and did not affect maternal survival, suggesting a conservative strategy. Disentangling within- and between-individual responses revealed trade-offs within individuals, but because individuals did not vary in their responses to earlier effort, these trade-offs did not drive population trends. The interacting effects of environmental conditions, maternal characteristics and individual variation on allocation trade-offs demonstrate the importance of long-term monitoring for understanding life history variations in changing environments.

© 2019, Springer-Verlag GmbH Germany, part of Springer Nature. Purpose: Animal models are frequently used to elucidate pathomechanism and pathophysiology of various disorders of the human intervertebral disc (IVD) and also to develop therapeutic approaches. Here we report morphological characteristics of the kangaroo lumbar IVDs and compare them with other animal models used in spine research. Methods: Twenty-five fresh-frozen cadaveric lumbar spines (T12–S1) derived from kangaroo carcases (Macropus giganteus) of undetermined age were first scanned in a C-Arm X-ray machine. A photograph of the axial section of the disc including a calibrated metric scale was also acquired. The digital radiographs and photographs were processed in ImageJ to determine the axial and sagittal plane dimensions for the whole disc (WD) and the nucleus pulposus (NP) and the mid-sagittal disc height for all the lumbar levels. Results: Our results suggest that the L6–S1 IVD in kangaroos is distinctly large compared with the upper lumbar IVDs. Based on previously published data, human lumbar IVDs are the largest of all the animal IVDs used in spine research, with camelid cervical IVDs being the closest relative in absolute dimensions (llamas: 78% in disc height, 40% in WD volume, and 38% in NP volume). Kangaroo L6–S1 IVD was approximately 51% in height, 20% in WD volume, and 20% in NP volume of the human lumbar IVD. Conclusions: We conclude that morphological similarities exist between a kangaroo and human lumbar IVD, especially with the lima bean shape in the axial plane, wedge shape in the sagittal plane, convexity at the cephalad endplates, and percentage volume occupied by the NP in the IVD. Graphic abstract: These slides can be retrieved under Electronic Supplementary Material. [Figure not available: see fulltext.]

Genomic imprinting is found in marsupial and eutherian mammals, but not in monotremes. While the primary regulator of genomic imprinting in eutherians is differential DNA methylation between parental alleles, conserved imprinted genes in marsupials tend to lack DNA methylation at their promoters. DNA methylation at eutherian imprinted genes is mainly catalyzed by a DNA methyltransferase (DNMT) enzyme, DNMT3A. There are two isoforms of eutherian DNMT3A: DNMT3A and DNMT3A2. DNMT3A2 is the primary isoform for establishing DNA methylation at eutherian imprinted genes and is essential for eutherian genomic imprinting. In this study, we investigated whether DNMT3A2 is also present in the two other mammalian lineages, marsupials and monotremes. We identified DNMT3A2 in both marsupials and monotremes, although imprinting has not been identified in monotremes. By analyzing genomic sequences and transcriptome data across vertebrates, we concluded that the evolution of DNMT3A2 occurred in the common ancestor of mammals. In addition, DNMT3A/3A2 gene and protein expression during gametogenesis showed distinct sexual dimorphisms in a marsupial, the tammar wallaby, and this pattern coincided with the sex-specific DNA methylation reprogramming in this species as it does in mice. Our results show that DNMT3A2 is present in all mammalian groups and suggests that the basic DNMT3A/3A2-based DNA methylation mechanism is conserved at least in therian mammals.

Specimen identification is the backbone of archeozoological research. The challenge of differentiating postcranial skeletal elements of closely related wild animals in biodiverse regions can prove a barrier to understanding past human foraging behaviours. Morphometrics are increasingly being employed to classify paleozoological animal remains, however, the potential of these methods to discriminate between wild animal groups has yet to be fully realised. Here we demonstrate the applicability of a traditional morphometric approach to taxonomically classify foot and ankle bones of kangaroos, a large and highly diverse marsupial family. Using multiple discriminant analysis, we classify archaeological specimens from Boodie Cave, in northwest Australia and identify the presence of two locally extinct macropod species during the terminal Pleistocene. The appearance of the banded hare-wallaby and northern nail-tail wallaby in the Pilbara region at this time provides independent evidence of the ecological and human responses to a changing climate at the end of the last Ice Age. Traditional morphometrics provides an accessible, inexpensive, and non-destructive tool for paleozoological specimen classification and has substantial potential for applications to other diverse wild faunas.

Background The eutherian IGF2R imprinted domain is regulated by an antisense long non-coding RNA, Airn, which is expressed from a differentially methylated region (DMR) in mice. Airn silences two neighbouring genes, Solute carrier family 22 member 2 (Slc22a2) and Slc22a3, to establish the Igf2r imprinted domain in the mouse placenta. Marsupials also have an antisense non-coding RNA, ALID, expressed from a DMR, although the exact function of ALID is currently unknown. The eutherian IGF2R DMR is located in intron 2, while the marsupial IGF2R DMR is located in intron 12, but it is not yet known whether the adjacent genes SLC22A2 and/or SLC22A3 are also imprinted in the marsupial lineage. In this study, the imprinting status of marsupial SLC22A2 and SLC22A3 in the IGF2R imprinted domain in the chorio-vitelline placenta was examined in a marsupial, the tammar wallaby. Results In the tammar placenta, SLC22A3 but not SLC22A2 was imprinted. Tammar SLC22A3 imprinting was evident in placental tissues but not in the other tissues examined in this study. A putative promoter of SLC22A3 lacked DNA methylation, suggesting that this gene is not directly silenced by a DMR on its promoter as seen in the mouse. Based on immunofluorescence, we confirmed that the tammar SLC22A3 is localised in the endodermal cell layer of the tammar placenta where nutrient trafficking occurs. Conclusions Since SLC22A3 is imprinted in the tammar placenta, we conclude that this placental imprinting of SLC22A3 has been positively selected after the marsupial and eutherian split because of the differences in the DMR location. Since SLC22A3 is known to act as a transporter molecule for nutrient transfer in the eutherian placenta, we suggest it was strongly selected to control the balance between supply and demand of nutrients in marsupial as it does in eutherian placentas.

With little research investigating older Australians' experience of companion animal (pet) ownership or wildlife encounters, this paper focuses on the value of animals in their lives.Three online focus groups explored 103 older adults' lived experience of home and community, with this paper focused on interactions with animals-companion animals and wildlife.Most (60%) had at least one pet, which brought love and joy into their lives. Reflexive thematic analysis identified three themes: pets as family; challenges of pet ownership; and connection to everyday wildlife and nature.In addition to valuing companion animals (typically cats and dogs), this research is among the first to show that some older people also develop ongoing relationships with local Australian wildlife, such as kookaburras, kangaroos, lizards and birds, which raises the possibility that fostering connections with wildlife may enhance well-being.

Wallabies are small- to medium-sized hopping marsupials and have large and flexible tendons in their hind limbs that act like springs. This study aimed to show the morphological pattern of the pes tendons in Bennett's wallaby. Two Bennett's Wallabies (Macropus rufogriseus) that died of natural causes have been used for this study. The pes was dissected using standard dissection techniques to expose the tendons around metatarsals and digits. The crural musculature of the hind limb was also dissected to identify the origin of the tendons. Tendons of m. extensor digitorum longus, m. extensor digitorum lateralis, m. extensor digiti II et III longus, m. flexor digitorum superficialis, m. flexor digitorum profundus and mm. interossei were the main identified tendons. Tendons of m. extensor digitorum longus attached to the distal phalanx of the fourth digit. The tendon of m. extensor digitorum lateralis had two insertion points, on the fourth and the fifth digits. The tendon of m. flexor digitorum superficialis bifurcates at the level proximal one-third of the metatarsus. The relatively thinner branch inserted into the phalanx of the fifth digit, while the thicker splits and inserted to the medial and lateral surface of the distal end of the proximal phalanx of the fourth digit. Tendon of m. flexor digitorum profundus was the thickest tendon on the plantar surface, and it had four insertion points, which were the distal phalanges of the second, third, fourth and fifth digits. This study provides detailed information for future studies on the biomechanical and functional morphology of tendons in marsupials.

During meiotic prophase I, homologous chromosomes pair, synapse and recombine in a tightly regulated process that ensures the generation of genetically variable haploid gametes. Although the mechanisms underlying meiotic cell division have been well studied in model species, our understanding of the dynamics of meiotic prophase I in non-traditional model mammals remains in its infancy. Here, we reveal key meiotic features in previously uncharacterised marsupial species (the tammar wallaby and the fat-tailed dunnart), plus the fat-tailed mouse opossum, with a focus on sex chromosome pairing strategies, recombina tion and meiotic telomere homeostasis. We uncovered differences between phylogroups with important functional and evolutionary implications. First, sex chromosomes, which lack a pseudo-autosomal region in marsupials, had species specific pairing and silencing strategies, with implications for sex chromosome evolution. Second, we detected two waves of γH2AX accumulation during prophase I. The first wave was accompanied by low γH2AX lev els on autosomes, which correlated with the low recombination rates that distinguish marsupials from eutherian mammals. In the second wave, γH2AX was restricted to sex chromosomes in all three species, which correlated with transcription from the X in tammar wallaby. This suggests non-canonical functions of γH2AX on meiotic sex chromosomes. Finally, we uncover evidence for telomere elongation in primary spermatocytes of the fattailed dunnart, a unique strategy within mammals. Our results provide new insights into meiotic progression and telomere homeostasis in marsupials, highlighting the importance of capturing the diversity of meiotic strategies within mammals, This work was supported by the Ministry of Economy, Industry and Competitiveness (CGL2017-83802-P to A.R-H. and CGL2014- 53106-P to J.P.), the Spanish Ministry of Science and Innovation (PID2020-112557GB-I00 to A.R H.) and the Agència de Gestio´ d’Ajuts Universitaris i de Recerca, AGAUR (SGR1215 to A.R-H.)

Prominent ontogenetic changes of the gastrointestinal tract (GIT) should occur in mammals whose neonatal diet of milk differs from that of adults, and especially in herbivores (as vegetation is particularly distinct from milk), and even more so in foregut fermenters, whose forestomach only becomes functionally relevant with vegetation intake. Due to the protracted lactation in marsupials, ontogenetic differences can be particularly well investigated in this group. Here, we report body mass (BM) scaling relationships of wet GIT content mass in 28 in-pouch young (50 g to 3 kg) and 15 adult (16–70 kg) western grey kangaroos Macropus fuliginosus melanops. Apart from the small intestinal contents, in-pouch young and adults did not differ in the scaling exponents (‘slope’ in log-log plots) but did differ in the scaling factor (‘intercept’), with an implied substantial increase in wet GIT content mass during the out-of-pouch juvenile period. In contrast to forestomach contents, caecum contents were elevated in juveniles still in the pouch, suggestive of fermentative digestion of milk and intestinal secretion residues, particularly in the caecum. The substantial increase in GIT contents (from less than 1 to 10–20% of BM) was associated mainly with the increase in forestomach contents (from 25 to 80% of total GIT contents) and a concomitant decrease in small intestine contents (from 50 to 8%), emphasizing the shifting relevance of auto-enzymatic and allo-enzymatic (microbial) digestion. There was a concomitant increase in the contents-to-tissue ratio of the fermentation chambers (forestomach and caecum), but this ratio generally did not change for the small intestine. Our study not only documents significant ontogenetic changes in digestive morpho-physiology, but also exemplifies the usefulness of intraspecific allometric analyses for quantifying these changes.

Xanthomas are localized lipid deposits in organs with associated granulomatous inflammation. Xanthomatosis is a rare condition in both human and veterinary medicine and is often linked to inherited or acquired dyslipidemias. Three female yellow-footed rock wallabies (

CASE DESCRIPTION A 7-month-old hand-reared female Bennett’s wallaby (Macropus rufogriseus) was evaluated for bilateral ocular opacity of 3 months’ duration. CLINICAL FINDINGS On physical examination, the wallaby was in good overall condition. An ophthalmic examination revealed mature cataracts in both eyes (OU). The cataracts were suspected to have a nutritional origin as it has been described in hand-reared macropods. Results of tonometry were normal OU. Results of CBC and serum biochemistry were unremarkable. The wallaby was premedicated with medetomidine and ketamine hydrochloride. Ocular ultrasonography performed while the wallaby was anesthetized revealed a thickened and opaque lens, mildly heterogeneous vitreous humor, and no sign of retinal detachment OU. An electroretinogram confirmed functional retinae OU. TREATMENT AND OUTCOME Surgery was considered necessary for welfare reasons. Phacoemulsification and vitrectomy were performed without intraoperative complication. The wallaby recovered uneventfully, and systemic NSAID, topical corticosteroid, and systemic and topical antimicrobial therapy were prescribed. One year later, the animal had vision OU. Physical examination did not show any vision-threatening postoperative complications. CLINICAL RELEVANCE The etiology of nutritional cataracts is not fully understood in macropods, but this condition is rather common in these species. To our knowledge, this is the first detailed report of a successful bilateral phacoemulsification and vitrectomy in a Bennett’s wallaby.

Chromosome rearrangements can result in the rapid evolution of hybrid incompatibilities. Robertsonian fusions, particularly those with monobrachial homology, can drive reproductive isolation amongst recently diverged taxa. The recent radiation of rock-wallabies (genus Petrogale) is an important model to explore the role of Robertsonian fusions in speciation. Here, we pursue that goal using an extensive sampling of populations and genomes of Petrogale from north-eastern Australia. In contrast to previous assessments using mitochondrial DNA or nuclear microsatellite loci, genomic data are able to separate the most closely related species and to resolve their divergence histories. Both phylogenetic and population genetic analyses indicate introgression between two species that differ by a single Robertsonian fusion. Based on the available data, there is also evidence for introgression between two species which share complex chromosomal rearrangements. However, the remaining results show no consistent signature of introgression amongst species pairs and where evident, indicate generally low introgression overall. X-linked loci have elevated divergence compared with autosomal loci indicating a potential role for genic evolution to produce reproductive isolation in concert with chromosome change. Our results highlight the value of genome scale data in evaluating the role of Robertsonian fusions and structural variation in divergence, speciation, and patterns of molecular evolution.

The evolution of Australia’s distinctive marsupial fauna has long been linked to the onset of continent-wide aridity. However, how this profound climate change event affected the diversification of extant lineages is still hotly debated. Here, we assemble a DNA sequence dataset of Macropodoidea—the clade comprising kangaroos and their relatives—that incorporates a complete mitogenome for the Desert ‘rat-kangaroo’, Caloprymnus campestris. This enigmatic species went extinct nearly 90 years ago and is known from a handful of museum specimens. Caloprymnus is significant because it was the only macropodoid restricted to extreme desert environments, and therefore calibrates the group’s specialisation for increasingly arid conditions. Our robustly supported phylogenies nest Caloprymnus amongst the bettongs Aepyprymnus and Bettongia. Dated ancestral range estimations further reveal that the Caloprymnus-Bettongia lineage originated in nascent xeric settings during the middle to late Miocene, ~ 12 million years ago (Ma), but subsequently radiated into fragmenting mesic habitats after the Pliocene to mid-Pleistocene. This timeframe parallels the ancestral divergences of kangaroos in woodlands and forests, but predates their adaptive dispersal into proliferating dry shrublands and grasslands from the late Miocene to mid-Pleistocene, after ~ 7 Ma. We thus demonstrate that protracted changes in both climate and vegetation likely staged the emergence of modern arid zone macropodoids.

To date, more than 50 Eimeria spp. have been isolated from marsupials of the family Macropodidae. Although 18 species of Eimeria have been previously detected from multiple animal species belonging to the genus Macropus of the family, limited genetic analyses of the parasites are available, and their pathogenicity remains unclear. Here, we report the isolation of Eimeria spp. from a zoo specimen of red-necked wallaby (Macropodidae; Macropus rufogriseus). Specifically, two distinct types of Eimeria oocysts were recovered, one from the feces before treatment with an anthelmintic and the second from the intestinal contents after death of the animal. The oocysts obtained from the two sources were morphologically identified as E. hestermani and E. prionotemni, respectively. We successfully determined partial gene sequences from the two isolates, including segments of the 18S rRNA genes, and for the first time have used phylogenetic analyses of these sequences to assign the species to distinct clades. In combination with further genetic data, these results are expected to help elucidate the pathogenicity and host ranges of Eimeria spp. within the respective family and genus.

Two new species of Wallabinema Beveridge, 1983 are described from the sacculated fore-stomachs of macropodid marsupials in Australia. Wallabinema petrogale n. sp. from the rock wallabies Petrogale penicillata (J. Gray) and P. inornata Gould, from Queensland, differs from all congeners in having its four sub-median lips subdivided at the base. Wallabinema macropodis n. sp. from the black stripe wallaby Macropus dorsalis (J. Gray) and the red-necked pademelon Thylogale thetis (Lesson), also from Queensland, is most similar to congeners with the nerve-ring encircling the oesophageal isthmus (W. parvispiculare Beveridge, 1983, W. tasmaniense Beveridge, 1983 and W. thylogale Beveridge, 1983), but differs in the length of the spicules. Wallabinema gallardi (Johnston & Mawson, 1939) is redescribed from T. thetis in Queensland and New South Wales, with T. thetis considered to be the type-host.

Four new species of Rugopharynx Monnig, 1927 are described from macropodid marsupials in Australia, some identified initially using molecular methods. Rugopharynx thetidis n. sp. from Thylogale thetis (Lesson) was initially included within R. sigma Chilton, Beveridge & Andrews, 1993 but was identified as being distinctive using molecular methods and is differentiated morphologically from R. sigma by the more anterior position of the deirid and by the distinctive curvature of the spicule tips. Rugopharynx thylogale n. sp., also from Thylogale thetis, is distinguished by the extreme elongation of the dorsal lobe of the bursa and spicule length (1.07-1.23 mm). Rugopharynx solitarius n. sp., again from Thylogale thetis, was initially included within Rugopharynx zeta (Johnston & Mawson, 1939) but differs morphologically in the features of the bursa. Rugopharynx disiunctus n. sp. from Macropus fuliginosus (Desmarest) was initially included within Rugopharynx rho Beveridge & Chilton, 1999 but was identified as distinctive based on DNA sequence differences. The new species differs from R. rho in the pattern of striations on the buccal capsule and in the lengths of the spicules. Additional records of species of Rugopharyx in M. fuliginosus in Western Australia are provided.

Objective: A large-scale capture method was developed to enable sterilisation of a macropod population in western Sydney from 2005 to 2018. Methods: Until March 2007, free ranging eastern grey kangaroos and red kangaroos were herded into purpose-built 15 m diameter capture yards (CYs) for darting with a projectile syringe. From March 2007 onwards, animals were free-range darted in large areas without herding. Kangaroos were darted with 1.33–5.10 mg/kg tiletamine/zolazepam and 0.01–0.02 mg/kg medetomidine, ± 0.03 mg/kg acepromazine. Deaths were monitored. Population counts were performed annually. Results: There were 5825 capture events involving 3963 kangaroos. Over 85% of all captures occurred from 2005 to 2008. Of all reported deaths (n = 523), 135 were attributed to ill health. Musculoskeletal injuries incurred during capture were the main project-related cause of death (n = 116). Post capture myopathy was uncommonly diagnosed following capture (n = 19). Conclusion: The herding and capture method enabled a large number of kangaroos to be mobilised and captured with low mortality rates, and the use of CYs resulted in fewer capture-related injuries and deaths than free-range capture. The drug doses and combinations used for darting were safe and effective, and the capture technique was successfully applied to a population management project.

A 1-year-old male red kangaroo (Macropus rufus) with an acute clinical history of lethargy, depression and increased respiratory rate was presented for necropsy examination. Gross lesions in the digestive tract were a distended stomach with watery content and multifocal raised ulcers covered by fibrinous pseudomembranes in the forestomach and caecum. On histopathology, there was necrotizing and ulcerative gastritis and typhlitis with intralesional amoebic trophozoites and Toxoplasma gondii zoites. Lesions due to T. gondii infection were observed in multiple organs and diagnosis was confirmed by fluorescent antibody test and immunohistochemistry. Both toxoplasmosis and gastric amoebiasis are diseases described in macropods. In this case report, we describe concurrent disease caused by both protozoa in a red kangaroo. Lesions of amoebiasis were also observed in the caecum. Both toxoplasmosis and amoebiasis should be considered as differential diagnoses of ulcerative lesions in the gastrointestinal tract in macropods during necropsy examination. Amoebiasis should be suspected especially when ulcerative lesions are observed in the forestomach.

A red-necked wallaby (Macropus rufogriseus) in the Republic of Korea, introduced from Australia, died in 12 d after exhibiting anorexia and diarrhea. Postmortem examination revealed that the wallaby died due to coccidiosis by Eimeria sp. Morphologically, Eimeria sp. identified closely resembled E. mykytowyczi. The 18S rRNA sequence analysis showed that Eimeria sp. identified in this study has a 98.5% identity with that in Australian red kangaroo (M. rufus). However, owing to insufficient molecular information on marsupial-specific Eimeria, exact species could not be determined. Phylogenetically, Eimeria sp. identified in this study belonged to clade five of the marsupial group.

OBJECTIVE Non-fatal injuries sustained from animal-vehicle collisions are a globally under-recognised road safety issue, with limited data on these crash types. The present study aimed to quantify the number and causes of major trauma events resulting from animal-vehicle collisions. METHODS The study was a retrospective analysis of major trauma cases occurring in Victoria, Australia, between 2007 and 2016, using data from the population-based Victorian State Trauma Registry. To identify animal-vehicle collisions, Victorian State Trauma Registry injury codes were combined with text-mining of the text description of the injury event. RESULTS Over the 10 year period, there were 152 major trauma patients who were admitted to Victorian trauma-receiving hospitals due to vehicle collisions with animals. The crude population-based incidence rate for animal-vehicle collisions increased by 6.7% per year (incidence rate ratio 1.07; 95% confidence interval 1.01-1.13; P = 0.02). CONCLUSION Development of systematic recording methods of animal-vehicle collisions will improve reporting of these crash types to assist future studies in implementing effective countermeasures.

The helminth parasites of the western scrub wallaby or black-glove wallaby, Notamacropus irma (Jourdan) which occurs in Western Australia are relatively poorly documented. Six new species of the strongyloid genus Cloacina von Linstow, 1898 (Strongylida: Chabertiidae) are described namely C. asymmetrica n. sp., C. brazellei n. sp., C. harriganae n. sp., C. hobbsi n. sp., C. middletoni n. sp. and C. woodi n. sp. A redescription of C. laius Beveridge, 1999 from the same host species is included. Molecular sequence data (ITS1 and ITS2 ribosomal DNA) were obtained for C. asymmetrica, C. brazellei, C. hobbsi, C. middletoni and from the previously described species C. themis Beveridge, 1998 occurring in the same host species. Phylogenetically, C. asymmetrica, C. hobbsi and C. middletoni formed a distinct clade, suggesting the possibility of within-host speciation. Cloacina themis clustered with a group of morphologically distinctive species in a separate clade and C. brazellei clustered in a third clade but with poor support. This pattern of congeners in a single host species occurring in multiple clades mirrors the situation in other kangaroos and wallabies. Species of Cloacina from N. irma reported thus far therefore consist of a series of species found only in this host, with two species (C. brazellei and C. laius) shared with the sympatric macropodid Setonix brachyurus (Quoy & Gaimard).

The objectives of this clinical study were to compare the effectiveness and safety of medetomidine-ketamine-midazolam (MKM) versus medetomidine-ketamine-butorphanol (MKB) for immobilization of captive red kangaroos (