Characterising the Tasmanian devil (Sarcophilus harrisii) pouch microbiome in lactating and non-lactating females.

Wildlife harbour a diverse range of microorganisms that affect their health and development. Marsupials are born immunologically naïve and physiologically underdeveloped, with primary development occurring inside a pouch. Secretion of immunological compounds and antimicrobial peptides in the epithelial lining of the female's pouch, pouch young skin, and through the milk, are thought to boost the neonate's immune system and potentially alter the pouch skin microbiome. Here, using 16S rRNA amplicon sequencing, we characterised the Tasmanian devil pouch skin microbiome from 25 lactating and 30 non-lactating wild females to describe and compare across these reproductive stages. We found that the lactating pouch skin microbiome had significantly lower amplicon sequence variant richness and diversity than non-lactating pouches, however there was no overall dissimilarity in community structure between lactating and non-lactating pouches. The top five phyla were found to be consistent between both reproductive stages, with over 85% of the microbiome being comprised of Firmicutes, Proteobacteria, Fusobacteriota, Actinobacteriota, and Bacteroidota. The most abundant taxa remained consistent across all taxonomic ranks between lactating and non-lactating pouch types. This suggests that any potential immunological compounds or antimicrobial peptide secretions did not significantly influence the main community members. Of the more than 16,000 total identified amplicon sequence variants, 25 were recognised as differentially abundant between lactating and non-lactating pouches. It is proposed that the secretion of antimicrobial peptides in the pouch act to modulate these microbial communities. This study identifies candidate bacterial clades on which to test the activity of Tasmanian devil antimicrobial peptides and their role in pouch young protection, which in turn may lead to future therapeutic development for human diseases.

Density of a cryptic Australian small mammal: The threatened Julia Creek dunnart (Sminthopsis douglasi).

Globally, hundreds of mammal species face the threat of extinction in the coming decades, and in many cases, their ecology remains poorly understood. Fundamental ecological knowledge is crucial for effective conservation management of these species, but it is particularly lacking for small, cryptic mammals. The Julia Creek dunnart (Sminthopsis douglasi), a threatened, cryptic carnivorous marsupial that occurs in scattered populations in the central west of Queensland, Australia, was once so poorly studied that it was believed extinct. Sporadic research since its rediscovery in the early 1990s has revealed that S. douglasi is distributed across land at risk from many threats. Fundamental knowledge of S. douglasi population density is urgently required to inform conservation management at key sites, yet the species has historically proven hard to detect. Indeed, the status of the largest known population of S. douglasi, in Bladensburg National Park, is unknown. Here, we conducted a population study on S. douglasi at two sites within Bladensburg National Park via live mark-recapture surveys during 2022 and 2023. From likelihood-based spatially explicit capture-recapture (SECR) modelling we provide the first estimates of density and population size for S. douglasi. Live trapping resulted in captures of 49 individual S. douglasi (with 83 captures total, including recaptures). We estimated S. douglasi to occur at a density of 0.38 individuals ha-1 (0.25-0.58) at one site and 0.16 individuals ha-1 (0.09-0.27) at another site, with an estimated mean population size in suitable habitat at Bladensburg National Park of 1211 individuals (776-1646). Our S. douglasi density estimates were similar to that reported for other threatened small mammals in Australia. We also found evidence of extreme S. douglasi population fluctuations over time at Bladensburg National Park, which is of concern for its future conservation. Our study has provided the first estimate of density for S. douglasi, a threatened dasyurid species from the Mitchell Grass Downs of central western Queensland, Australia. Our research provides crucial population data to assist the management of this poorly studied species. We demonstrate a method that can be applied to species with low detection probability to ultimately help address the mammal extinction crisis faced by Australia and the rest of the world.

Development of the pulmonary vasculature in the gray short-tailed opossum (Monodelphis domestica)-3D reconstruction by microcomputed tomography.

In the marsupial gray short-tailed opossum (Monodelphis domestica), the majority of lung development, including the maturation of pulmonary vasculature, takes place in ventilated functioning state during the postnatal period. The current study uses X-ray computed tomography (µCT) to three-dimensionally reconstruct the vascular trees of the pulmonary artery and pulmonary vein in 15 animals from neonate to postnatal day 57. The final 3D reconstructions of the pulmonary artery and pulmonary vein in the neonate and at 21, 35, and 57 dpn were transformed into a centerline model of the vascular trees. Based on the reconstructions, the generation of end-branching vessels, the median and maximum generation, and the number of vessels were calculated for the lungs. The pulmonary vasculature follows the lung anatomy with six pulmonary lobes indicated by the bronchial tree. The pulmonary arteries follow the bronchial tree closely, in contrast to the pulmonary veins, which run between the pulmonary segments. At birth the pulmonary vasculature has a simple branching pattern with a few vessel generations. Compared with the bronchial tree, the pulmonary vasculature appears to be more developed and extends to the large terminal air spaces. The pulmonary vasculature shows a marked gain in volume and a progressive increase in vascular complexity and density. The gray short-tailed opossum resembles the assumed mammalian ancestor and is suitable to inform on the evolution of the mammalian lung. Vascular genesis in the marsupial bears resemblance to developmental patterns described in eutherians. Lung development in general seems to be highly conservative within mammalian evolution.

Lead exposure and source attribution for a mammalian scavenger before and after a culling program.

Lead-based ammunition is a significant source of environmental lead and threatens species that scavenge lead-shot carcasses, particularly in areas with intensive shooting. With the impacts of lead on avian scavengers well established, there is increasing focus on the effects of lead on mammalian scavengers. We investigated lead exposure in a morphologically specialized mammalian scavenger, the Tasmanian devil (Sarcophilus harrisii), by analyzing their blood lead levels (BLLs) before and after a marsupial culling program using linear mixed effects models. We compared lead isotope signatures in devil blood to those in the culling ammunition to inform potential source attributions. We sampled 23 devils before culling and 15 after culling, finding no significant difference in mean BLLs pre and post-culling. However, devils captured closer to forestry coupes where culling had occurred had higher BLLs, and a greater proportion of devils displayed elevated BLLs post-culling (33 % compared to 18 % pre-culling). The highest BLL (7.93 µg/dL) was found in a devil post-culling and this individual had lead isotope signatures that matched the ammunition samples analyzed, suggesting the individual was exposed to lead from scavenging on culled carcasses. While 18 % of the devil blood lead samples had isotope signatures consistent with the ammunition samples, most were measurably different, indicating other sources of lead in the landscape. BLLs in our study landscape were similar to published BLLs for wild devils across Tasmania. That said, lead isotope signatures in the blood of individual devils sampled both before and after culling shifted closer to those of ammunition samples post-culling. Our results indicate that while some individual devils may have been exposed to lead from culling, most devils in the landscape did not show evidence of recent exposure. However, even low lead levels can adversely impact wildlife health and immunity, a particular concern for devils, a species endangered by disease.

Embryology of the fat-tailed dunnart (Sminthopsis crassicaudata): A marsupial model for comparative mammalian developmental and evolutionary biology.

BACKGROUND: Marsupials are a diverse and unique group of mammals, but remain underutilized in developmental biology studies, hindering our understanding of mammalian diversity. This study focuses on establishing the fat-tailed dunnart (Sminthopsis crassicaudata) as an emerging laboratory model, providing reproductive monitoring methods and a detailed atlas of its embryonic development. RESULTS: We monitored the reproductive cycles of female dunnarts and established methods to confirm pregnancy and generate timed embryos. With this, we characterized dunnart embryo development from cleavage to birth, and provided detailed descriptions of its organogenesis and heterochronic growth patterns. Drawing stage-matched comparisons with other species, we highlight the dunnarts accelerated craniofacial and limb development, characteristic of marsupials. CONCLUSIONS: The fat-tailed dunnart is an exceptional marsupial model for developmental studies, where our detailed practices for reproductive monitoring and embryo collection enhance its accessibility in other laboratories. The accelerated developmental patterns observed in the Dunnart provide a valuable system for investigating molecular mechanisms underlying heterochrony. This study not only contributes to our understanding of marsupial development but also equips the scientific community with new resources for addressing biodiversity challenges and developing effective conservation strategies in marsupials.

Sex and landscape influence spatial genetic variation in a large fossorial mammal, the Bare-nosed Wombat (Vombatus ursinus).

Dispersal is an important process that is widely studied across species, and it can be influenced by intrinsic and extrinsic factors. Intrinsic factors commonly assessed include the sex and age of individuals, while landscape features are frequently-tested extrinsic factors. Here, we investigated the effects of both sex and landscape composition and configuration on genetic distances among bare-nosed wombats (Vombatus ursinus)-one of the largest fossorial mammals in the world and subject to habitat fragmentation, threats from disease, and human persecution including culling as an agricultural pest. We analyzed a data set comprising 74 Tasmanian individuals (30 males and 44 females), genotyped for 9,064 single-nucleotide polymorphisms. We tested for sex-biased dispersal and the influence of landscape features on genetic distances including land use, water, vegetation, elevation, and topographic ruggedness. We detected significant female-biased dispersal, which may be related to females donating burrows to their offspring due to the energetic cost of excavation, given their large body sizes. Land use, waterbodies, and elevation appeared to be significant landscape predictors of genetic distance. Land use potentially reflects land clearing and persecution over the last 200 years. If our findings based on a limited sample size are valid, retention and restoration of nonanthropogenic landscapes in which wombats can move and burrow may be important for gene flow and maintenance of genetic diversity.

Association of maternal genetics with the gut microbiome and eucalypt diet selection in captive koalas.

Background: Koalas, an Australian arboreal marsupial, depend on eucalypt tree leaves for their diet. They selectively consume only a few of the hundreds of available eucalypt species. Since the koala gut microbiome is essential for the digestion and detoxification of eucalypts, their individual differences in the gut microbiome may lead to variations in their eucalypt selection and eucalypt metabolic capacity. However, research focusing on the relationship between the gut microbiome and differences in food preferences is very limited. We aimed to determine whether individual and regional differences exist in the gut microbiome of koalas as well as the mechanism by which these differences influence eucalypt selection. Methods: Foraging data were collected from six koalas and a total of 62 feces were collected from 15 koalas of two zoos in Japan. The mitochondrial phylogenetic analysis was conducted to estimate the mitochondrial maternal origin of each koala. In addition, the 16S-based gut microbiome of 15 koalas was analyzed to determine the composition and diversity of each koala's gut microbiome. We used these data to investigate the relationship among mitochondrial maternal origin, gut microbiome and eucalypt diet selection. Results and Discussion: This research revealed that diversity and composition of the gut microbiome and that eucalypt diet selection of koalas differs among regions. We also revealed that the gut microbiome alpha diversity was correlated with foraging diversity in koalas. These individual and regional differences would result from vertical (maternal) transmission of the gut microbiome and represent an intraspecific variation in koala foraging strategies. Further, we demonstrated that certain gut bacteria were strongly correlated with both mitochondrial maternal origin and eucalypt foraging patterns. Bacteria found to be associated with mitochondrial maternal origin included bacteria involved in fiber digestion and degradation of secondary metabolites, such as the families Rikenellaceae and Synergistaceae. These bacteria may cause differences in metabolic capacity between individual and regional koalas and influence their eucalypt selection. Conclusion: We showed that the characteristics (composition and diversity) of the gut microbiome and eucalypt diet selection of koalas differ by individuals and regional origins as we expected. In addition, some gut bacteria that could influence eucalypt foraging of koalas showed the relationships with both mitochondrial maternal origin and eucalypt foraging pattern. These differences in the gut microbiome between regional origins may make a difference in eucalypt selection. Given the importance of the gut microbiome to koalas foraging on eucalypts and their strong symbiotic relationship, future studies should focus on the symbiotic relationship and coevolution between koalas and the gut microbiome to understand individual and regional differences in eucalypt diet selection by koalas.

Characterisation of defensins across the marsupial family tree.

Defensins are antimicrobial peptides involved in innate immunity, and gene number differs amongst eutherian mammals. Few studies have investigated defensins in marsupials, despite their potential involvement in immunological protection of altricial young. Here we use recently sequenced marsupial genomes and transcriptomes to annotate defensins in nine species across the marsupial family tree. We characterised 35 alpha and 286 beta defensins; gene number differed between species, although Dasyuromorphs had the largest repertoire. Defensins were encoded in three gene clusters within the genome, syntenic to eutherians, and were expressed in the pouch and mammary gland. Marsupial beta defensins were closely related to eutherians, however marsupial alpha defensins were more divergent. We identified marsupial orthologs of human DEFB3 and 6, and several marsupial-specific beta defensin lineages which may have novel functions. Marsupial predicted mature peptides were highly variable in length and sequence composition. We propose candidate peptides for future testing to elucidate the function of marsupial defensins.

DNA metabarcoding analysis of the bare-nosed wombat (Vombatus ursinus) diet.

Wombats are native herbivorous grazers that have adapted to Australia's low-quality forage. Studies on diet selection by bare-nosed wombats (Vombatus ursinus) are limited and are either observational or based on microhistological studies. The current study determined the diet of wombats through DNA metabarcoding across five study sites in New South Wales over a one-year period. Metabarcoding was chosen as it is non-invasive, less time consuming and more specific than traditional techniques. The list of 209 plant species identified as eaten by wombats in this study is much higher than previously reported, with grasses being the most common plant group identified in all samples. Most dietary items identified were introduced plant species. Seasonal differences in plants eaten occurred at four of the five study sites and may reflect dietary abundance and floristic composition at different times of year. Further studies are required to determine if the dietary items differ markedly across the entire range of wombats, and if nutrition influences dietary preferences.

Description of the parasitic fauna of a specimen of Didelphis albiventris at Rio Grande do Sul.

Didelphis albiventris is considered the most common marsupial in Rio Grande do Sul. With omnivorous and synanthropic habits, it can serve as a host to various parasites, playing an important role in maintaining their biological cycle. Despite being a widespread and abundant species, it has a relatively little-known parasitic fauna. Therefore, the aim of this study was to report the diversity of parasites in a fecal sample from D. albiventris in Rio Grande do Sul, Southern Brazil. Modified Centrifugal-flotation and Spontaneous sedimentation techniques were used, revealing a high taxonomic diversity of parasites. Eggs of Ancylostoma spp., Toxocara spp., and Anoplocephalidae were reported for the first time in the host in the southern region of the country, along with the first report of pseudoparasitism by Syphacia spp. and Monocystis spp. in this animal species. The presence of different parasites in the feces of D. albiventris is of utmost importance, primarily for public health, but also for understanding the biodiversity of parasites present in wildlife, which has been poorly studied until now. This allows the implementation of effective strategies for controlling, preventing and treating these diseases.

Didelphis albiventris é considerado o marsupial mais comum no Rio Grande do Sul. Com hábitos onívoros e sinantrópicos, pode servir de hospedeiro para diversos parasitas, desempenhando importante papel na manutenção do seu ciclo biológico. Apesar de ser uma espécie difundida e abundante, possui uma fauna parasitária relativamente pouco conhecida. Portanto, o objetivo deste estudo foi relatar a diversidade de parasitas em uma amostra fecal de D. albiventris no Rio Grande do Sul, Sul do Brasil. Foram utilizadas técnicas modificadas de flutuação centrífuga e sedimentação espontânea, revelando uma alta diversidade taxonômica de parasitas. Ovos de Ancylostoma spp., Toxocara spp. e Anoplocephalidae foram relatados pela primeira vez no hospedeiro na região sul do país, juntamente com o primeiro relato de pseudoparasitismo por Syphacia spp. e Monocystis spp. nesta espécie animal. A presença de diferentes parasitas nas fezes de D. albiventris é de extrema importância, principalmente para a saúde pública, mas também para a compreensão da biodiversidade de parasitas presentes na vida selvagem, que tem sido pouco estudada até agora. Isto permite a implementação de estratégias eficazes para controlar, prevenir e tratar estas doenças.

Spontaneous Lesions of Endangered Geriatric Julia Creek Dunnarts (Sminthopsis douglasi, Archer 1979) with Emphasis in Reproductive Pathology.

Julia Creek dunnarts are an endangered species of carnivorous marsupials and the focus of multiple conservation strategies involving significant resources such as captive breeding programs. Despite the relevance for conservation, no study to date has focused on evaluating geriatric diseases in dunnarts. This study describes the pathology findings in a group of one wild and thirty-five captive-born, mostly geriatric Julia Creek dunnarts that failed to produce offspring over multiple breeding periods. A total of 20 females and 16 males were submitted for a postmortem examination, with ages ranging from 9 to 42 and 12 to 42 months for females and males, respectively. Of these, 10 had unremarkable findings. The most common condition in females was cystic glandular hyperplasia (n = 8), typical of hormonal dysregulation profiles in senescence, particularly hyperestrogenism. Rarely, cutaneous disease represented by unidentified dermal round cell infiltrates was observed in females (n = 2). Primary reproductive hormonal dysregulation was also suspected in males diagnosed with testicular degeneration, aspermatogenesis and/or atrophy (n = 3). Cutaneous round cell infiltrates, possibly compatible with epitheliotropic lymphomas, were seen in males (n = 3), and 2/3 affected males also had concurrent testicular degeneration or atrophy, indicating male sex could be a predictor for lymphoid neoplasia in aged dunnarts, especially in individuals with concurrent testosterone-luteinizing hormone dysregulation as it occurs in gonadectomized animals. The role of an underlying viral etiology is also explored. This study is the first to describe major spontaneous diseases in endangered aged Julia Creek dunnarts, providing an important understanding of senescence and geriatric diseases within a conservation context.

Complex associations between cancer progression and immune gene expression reveals early influence of transmissible cancer on Tasmanian devils.

The world's largest extant carnivorous marsupial, the Tasmanian devil, is challenged by Devil Facial Tumor Disease (DFTD), a fatal, clonally transmitted cancer. In two decades, DFTD has spread across 95% of the species distributional range. A previous study has shown that factors such as season, geographic location, and infection with DFTD can impact the expression of immune genes in Tasmanian devils. To date, no study has investigated within-individual immune gene expression changes prior to and throughout the course of DFTD infection. To explore possible changes in immune response, we investigated four locations across Tasmania that differed in DFTD exposure history, ranging between 2 and >30 years. Our study demonstrated considerable complexity in the immune responses to DFTD. The same factors (sex, age, season, location and DFTD infection) affected immune gene expression both across and within devils, although seasonal and location specific variations were diminished in DFTD affected devils. We also found that expression of both adaptive and innate immune genes starts to alter early in DFTD infection and continues to change as DFTD progresses. A novel finding was that the lower expression of immune genes MHC-II, NKG2D and CD8 may predict susceptibility to earlier DFTD infection. A case study of a single devil with regressed tumor showed opposite/contrasting immune gene expression patterns compared to the general trends observed across devils with DFTD infection. Our study highlights the complexity of DFTD's interactions with the host immune system and the need for long-term studies to fully understand how DFTD alters the evolutionary trajectory of devil immunity.

Therian origin of INSL3/RXFP2-driven testicular descent in mammals.

Introduction: During early development in most male mammals the testes move from a position near the kidneys through the abdomen to eventually reside in the scrotum. The transabdominal phase of this migration is driven by insulin-like peptide 3 (INSL3) which stimulates growth of the gubernaculum, a key ligament connecting the testes with the abdominal wall. While all marsupials, except the marsupial mole (Notoryctes typhlops), have a scrotum and fully descended testes, it is unclear if INSL3 drives this process in marsupials especially given that marsupials have a different mechanism of scrotum determination and position relative to the phallus compared to eutherian mammals. Methods: To understand if INSL3 plays a role in marsupial testicular descent we have sequenced and curated the INSL3 gene and its receptor (RXFP2) in a range of marsupials representing every order. Furthermore, we looked at single cell RNA-seq and qPCR analysis of INSL3 in the fat-tailed dunnart testis (Sminthopsis crassicaudata) to understand the location and timing of expression during development. Results: These data show a strong phylogenetic similarity between marsupial and eutherian orthologues, but not with monotreme INSL3s which were more similar to the ancestral RLN3 gene. We have also shown the genomic location of INSL3, and surrounding genes is conserved in a range of marsupials and eutherians. Single cell RNA-seq and qPCR data show that INSL3 mRNA is expressed specifically in Leydig cells and expressed at higher levels during the testicular descent phase in developing marsupials. Discussion: Together, these data argue strongly for a therian origin of INSL3 mediated testicular descent in mammals and suggests that a coordinated movement of the testes to the abdominal wall may have preceded externalization in marsupials and therian mammals.

Astrocytes of the Anterior Commissure Regulate the Axon Guidance Pathways of Newly Generated Neocortical Neurons in the Opossum Monodelphis domestica.

In marsupials, upper-layer cortical neurons derived from the progenitors of the subventricular zone of the lateral ventricle (SVZ) mature morphologically and send their axons to form interhemispheric connections through the anterior commissure. In contrast, eutherians have evolved a new extra callosal pathway, the corpus callosum, that interconnects both hemispheres. In this study, we aimed to examine neurogenesis during the formation of cortical upper layers, including their morphological maturation in a marsupial species, namely the opossum (Monodelphis domestica). Furthermore, we studied how the axons of upper layers neurons pass through the anterior commissure of the opossum, which connects neocortical areas. We showed that upper-layer II/III neurons were generated within at least seven days in the opossum neocortex. Surprisingly, these neurons expressed special AT-rich sequence binding protein 2 (Satb2) and neuropilin 1 interacting protein (Nrp1), which are proteins known to be essential for the formation of the corpus callosum in eutherians. This indicates that extrinsic, but not intrinsic, cues could be key players in guiding the axons of newly generated cortical neurons in the opossum. Although oligodendrocyte precursor cells were present in the neocortex and anterior commissure, newly generated upper-layer neurons sent unmyelinated axons to the anterior commissure. We also found numerous GFAP-expressing progenitor cells in both brain structures, the neocortex and the anterior commissure. However, at P12-P17 in the opossums, a small population of astrocytes was observed only in the midline area of the anterior commissure. We postulate that in the opossum, midline astrocytes allow neocortical axons to be guided to cross the midline, as this structure resembles the glial wedge required by fibers to cross the midline area of the corpus callosum in the rodent.

Semelparous marsupials reduce sleep for sex.

Sleep is a prominent, seemingly universal animal behavior. Although sleep maintains optimal waking performance, the biological drive to sleep may be incompatible with the life history of some species. In a multi-year study on semelparous marsupials in Australia, we provide the first direct evidence of ecological sleep restriction in a terrestrial mammal. Dusky (Antechinus swainsonii) and agile (A. agilis) antechinus have an unusual reproductive strategy characterized by the synchronous death of all males at the end of their only breeding season. Using accelerometry, electrophysiology, and metabolomics, we show that males, but not females, increase their activity during the breeding season by reducing sleep. In a trade-off between the neurophysiological requirements for sleep and evolutionary necessity for reproduction, strong sexual selection might drive males to sacrifice sleep to increase access to fertile females and ultimately maximize their fitness.

Movement ecology of an endangered mesopredator in a mining landscape.

BACKGROUND: Efficient movement and energy expenditure are vital for animal survival. Human disturbance can alter animal movement due to changes in resource availability and threats. Some animals can exploit anthropogenic disturbances for more efficient movement, while others face restricted or inefficient movement due to fragmentation of high-resource habitats, and risks associated with disturbed habitats. Mining, a major anthropogenic disturbance, removes natural habitats, introduces new landscape features, and alters resource distribution in the landscape. This study investigates the effect of mining on the movement of an endangered mesopredator, the northern quoll (Dasyurus hallucatus). Using GPS collars and accelerometers, we investigate their habitat selection and energy expenditure in an active mining landscape, to determine the effects of this disturbance on northern quolls. METHODS: We fit northern quolls with GPS collars and accelerometers during breeding and non-breeding season at an active mine site in the Pilbara region of Western Australia. We investigated broad-scale movement by calculating the movement ranges of quolls using utilisation distributions at the 95% isopleth, and compared habitat types and environmental characteristics within observed movement ranges to the available landscape. We investigated fine-scale movement by quolls with integrated step selection functions, assessing the relative selection strength for each habitat covariate. Finally, we used piecewise structural equation modelling to analyse the influence of each habitat covariate on northern quoll energy expenditure. RESULTS: At the broad scale, northern quolls predominantly used rugged, rocky habitats, and used mining habitats in proportion to their availability. However, at the fine scale, habitat use varied between breeding and non-breeding seasons. During the breeding season, quolls notably avoided mining habitats, whereas in the non-breeding season, they frequented mining habitats equally to rocky and riparian habitats, albeit at a higher energetic cost. CONCLUSION: Mining impacts northern quolls by fragmenting favoured rocky habitats, increasing energy expenditure, and potentially impacting breeding dispersal. While mining habitats might offer limited resource opportunities in the non-breeding season, conservation efforts during active mining, including the creation of movement corridors and progressive habitat restoration would likely be useful. However, prioritising the preservation of natural rocky and riparian habitats in mining landscapes is vital for northern quoll conservation.

Dual-Responsive Nanorobot-Based Marsupial Robotic System for Intracranial Cross-Scale Targeting Drug Delivery.

Nanorobots capable of active movement are an exciting technology for targeted therapeutic intervention. However, the extensive motion range and hindrance of the blood-brain barrier impeded their clinical translation in glioblastoma therapy. Here, a marsupial robotic system constructed by integrating chemical/magnetic hybrid nanorobots (child robots) with a miniature magnetic continuum robot (mother robot) for intracranial cross-scale targeting drug delivery is reported. For primary targeting on macroscale, the continuum robot enters the cranial cavity through a minimally invasive channel (e.g., Ommaya device) in the skull and transports the nanorobots to pathogenic regions. Upon circumventing the blood-brain barrier, the released nanorobots perform secondary targeting on microscale to further enhance the spatial resolution of drug delivery. In vitro experiments against primary glioblastoma cells derived from different patients are conducted for personalized treatment guidance. The operation feasibility within organisms is shown in ex vivo swine brain experiments. The biosafety of the treatment system is suggested in in vivo experiments. Owing to the hierarchical targeting method, the targeting rate, targeting accuracy, and treatment efficacy have improved greatly. The marsupial robotic system offers a novel intracranial local therapeutic strategy and constitutes a key milestone in the development of glioblastoma treatment platforms.

Morphology and computed tomography of the larynx in the white-eared opossum (Didelphis albiventris).

The white-eared opossum (Didelphis albiventris) is a Neotropical marsupial that occurs in the Brazilian territory. The larynx is an important organ of vocalization in mammals, although, other laryngeal functions are more fundamental for survival of mammals than phonation. The anatomical knowledge of respiratory structures is pivotal for a better understanding of the species. Thus, this study aimed to examine the larynx of the white-eared opossum by gross anatomy, computed tomography and histological description. For this, 10 adult (six females and four males) white-eared opossums (D. albiventris) were used. The white-eared opossum larynx was formed by epiglottis, thyroid, cricoids and arytenoid cartilages and a corniculate process. There is a similarity between the larynx of this marsupial and those reported in other wild mammals, regarding the number of cartilages and their location. Histologically, the epiglottis consisted of elastic cartilage and thyroid, cricoid and arytenoid cartilages were composed of hyaline cartilage. The epiglottis protruded rostrally into the nasal part of the pharynx, above the soft palate, and this is probably a marsupial characteristic as the fact that the thyroid and cricoid cartilages were ventrally fused. The hyoid apparatus was similar to that of other animals, with the same bony constitution, but with a greatly reduced stylohyoid bone. Histologically, the larynx was similar to those of other species such as koala, armadillo, crab-eating foxes and giant anteaters. The knowledge of the larynx morphology is important for the anatomical features of the species and clinical and surgical procedures, such as endotracheal intubation.

Macropod Pediatrics.

Macropods belong to the marsupial family Macropodidae, which includes animals such as kangaroos and wallabies. Macropod offspring are highly altricial at birth and require specialized care and environmental conditions for healthy development. The care and management of pediatric macropods poses a challenge due to the unique physiology and reproductive strategy of macropods. In order to successfully work with pediatric macropods, clinical veterinarians should have knowledge of species-specific husbandry, normal postnatal development, and common medical conditions/treatments. With limited information available on macropod pediatric medicine, further research is warranted to improve the care and management of these animals in human care.

Sugar Glider Pediatrics.

Sugar gliders (Petaurus breviceps) are small nocturnal marsupials with a unique metabolism and reproductive strategy compared to other common exotic companion mammals. Although there are few reports on the pediatric diseases of sugar gliders, clinicians should be aware of the normal reproductive anatomy, physiology, and rearing of sugar gliders. By recognizing the signs of normal reproduction and joey development, clinicians can identify abnormalities quickly and provide appropriate intervention, as needed.