Distinctive diets of eutherian predators in Australia.

Introduction of the domestic cat and red fox has devastated Australian native fauna. We synthesized Australian diet analyses to identify traits of prey species in cat, fox and dingo diets, which prey were more frequent or distinctive to the diet of each predator, and quantified dietary overlap. Nearly half (45%) of all Australian terrestrial mammal, bird and reptile species occurred in the diets of one or more predators. Cat and dingo diets overlapped least (0.64 ± 0.27, n = 24 location/time points) and cat diet changed little over 55 years of study. Cats were more likely to have eaten birds, reptiles and small mammals than foxes or dingoes. Dingo diet remained constant over 53 years and constituted the largest mammal, bird and reptile prey species, including more macropods/potoroids, wombats, monotremes and bandicoots/bilbies than cats or foxes. Fox diet had greater overlap with both cats (0.79 ± 0.20, n = 37) and dingoes (0.73 ± 0.21, n = 42), fewer distinctive items (plant material, possums/gliders) and significant spatial and temporal heterogeneity over 69 years, suggesting the opportunity for prey switching (especially of mammal prey) to mitigate competition. Our study reinforced concerns about mesopredator impacts upon scarce/threatened species and the need to control foxes and cats for fauna conservation. However, extensive dietary overlap and opportunism, as well as low incidence of mesopredators in dingo diets, precluded resolution of the debate about possible dingo suppression of foxes and cats.

Intensive Adoption as a Management Strategy for Unowned, Urban Cats: A Case Study of 25 Years of Trap-Assess-Resolve (TAR) in Auckland, New Zealand.

Globally, unowned urban cats are a major concern because they may suffer from poor welfare and cause problems, including public health risks, nuisances, and urban wildlife predation. While management options are often presented as a choice between culling or trap−neuter−return (TNR), for 25 years, the Lonely Miaow (Inc.) charity in Auckland, New Zealand (hereafter LM), has used a third strategy­intensive adoption or trap−assess−resolve (TAR). As of 2019, of 14,611 unowned cats trapped, 64.2% were adopted, 22.2% were euthanized if unsocialised or in grave ill-health, 5.7% were neutered and returned to the site, and 7.9% had other outcomes, such as being transferred to other shelters. Adoption rates increased over this time, exceeding 80.0% in 2018 and 2019. The cost of processing each cat from capture to adoption rose from NZD 58 in 1999 to NZD 234 by 2017. Approximately 80% of colonies (sites where cats were trapped) were around residential areas. Approximately 22% of cats required veterinary treatment after capture; common ailments included respiratory infections, ringworm, dental problems, and trauma. Consistently, 52% of cats were young kittens (<10 weeks old), c. 80% of cats were <1 year old, and only c. 2% were estimated to be >5 years old. TAR avoids euthanasia where possible. Its effectiveness would be enhanced by fewer abandonments of owned cats and kittens, fitting within integrated strategies for the control of unowned cats involving community education. Cat adoptions improve the welfare of cats and, with appropriate husbandry, should alleviate concerns about nuisances, public health, and attacks on wildlife or the cats themselves, essentially benefitting the community and the cats. This case study is relevant to other cities around the world that are seeking to manage unowned cats.

A recurrent de novo ATP5F1A substitution associated with neonatal complex V deficiency.

Mitochondrial disorders are a heterogeneous group of rare, degenerative multisystem disorders affecting the cell's core bioenergetic and signalling functions. Spontaneous improvement is rare. We describe a novel neonatal-onset mitochondriopathy in three infants with failure to thrive, hyperlactatemia, hyperammonemia, and apparent clinical resolution before 18 months. Exome sequencing showed all three probands to be identically heterozygous for a recurrent de novo substitution, c.620G>A [p.(Arg207His)] in ATP5F1A, encoding the α-subunit of complex V. Patient-derived fibroblasts exhibited multiple deficits in complex V function and expression in vitro. Structural modelling predicts the observed substitution to create an abnormal region of negative charge on ATP5F1A's ß-subunit-interacting surface, adjacent to the nearby ß subunit's active site. This disorder, which presents with life-threatening neonatal manifestations, appears to follow a remitting course; the long-term prognosis remains unknown.

Histone H3.3 beyond cancer: Germline mutations in Histone 3 Family 3A and 3B cause a previously unidentified neurodegenerative disorder in 46 patients.

Although somatic mutations in Histone 3.3 (H3.3) are well-studied drivers of oncogenesis, the role of germline mutations remains unreported. We analyze 46 patients bearing de novo germline mutations in histone 3 family 3A (H3F3A) or H3F3B with progressive neurologic dysfunction and congenital anomalies without malignancies. Molecular modeling of all 37 variants demonstrated clear disruptions in interactions with DNA, other histones, and histone chaperone proteins. Patient histone posttranslational modifications (PTMs) analysis revealed notably aberrant local PTM patterns distinct from the somatic lysine mutations that cause global PTM dysregulation. RNA sequencing on patient cells demonstrated up-regulated gene expression related to mitosis and cell division, and cellular assays confirmed an increased proliferative capacity. A zebrafish model showed craniofacial anomalies and a defect in Foxd3-derived glia. These data suggest that the mechanism of germline mutations are distinct from cancer-associated somatic histone mutations but may converge on control of cell proliferation.

Body Size and Bite Force of Stray and Feral Cats-Are Bigger or Older Cats Taking the Largest or More Difficult-to-Handle Prey?

As carnivorans rely heavily on their head and jaws for prey capture and handling, skull morphology and bite force can therefore reflect their ability to take larger or more difficult-to-handle prey. For 568 feral and stray cats (Felis catus), we recorded their demographics (sex and age), source location (feral or stray) and morphological measures (body mass, body condition); we estimated potential bite force from skull measurements for n = 268 of these cats, and quantified diet composition from stomach contents for n = 358. We compared skull measurements to estimate their bite force and determine how it varied with sex, age, body mass, body condition. Body mass had the strongest influence of bite force. In our sample, males were 36.2% heavier and had 20.0% greater estimated bite force (206.2 ± 44.7 Newtons, n = 168) than females (171.9 ± 29.3 Newtons, n = 120). However, cat age was the strongest predictor of the size of prey that they had taken, with older cats taking larger prey. The predictive power of this relationship was poor though (r2 < 0.038, p < 0.003), because even small cats ate large prey and some of the largest cats ate small prey, such as invertebrates. Cats are opportunistic, generalist carnivores taking a broad range of prey. Their ability to handle larger prey increases as the cats grow, increasing their jaw strength, and improving their hunting skills, but even the smallest cats in our sample had tackled and consumed large and potentially 'dangerous' prey that would likely have put up a defence.

Response to Wolf et al.: Furthering Debate over the Suitability of Trap-Neuter-Return for Stray Cat Management.

To continue dialogue over proposed Australian trials of Trap-Neuter-Return (TNR), we applied a framework requiring identification of areas of agreement, areas of disagreement, and identification of empirical data collection required to resolve disagreements. There is agreement that Australia has a problem with stray cats, causing problems of impacts on wildlife, nuisance, disease transmission (including public health issues and exchange of diseases between stray cat and pet cat populations), poor welfare outcomes for stray cats, and an emotional burden on staff euthanising healthy stray cats. There is disagreement on whether (i) current measures are failing, leading to unacceptably high euthanasia levels, (ii) some contributors to the debate misunderstand TNR, (iii) TNR trials will reduce urban cat populations and associated problems, (iv) TNR is an ethical solution to cat overpopulation, and (v) some contributors to the debate promulgated misinformation. Although not everyone agrees that TNR trials should proceed, as a hypothetical exploration, we propose an experimental approach explicitly comparing TNR to alternatives. Trials could only be considered if other detailed and well-funded attempts at stray cat control focusing across an entire Local Government Area (LGA) prove ineffective.

A Case of Letting the Cat out of The Bag-Why Trap-Neuter-Return Is Not an Ethical Solution for Stray Cat (Felis catus) Management.

Trap-Neuter-Return (TNR) programs, in which stray cats are captured, neutered and returned to the environment are advocated as a humane, ethical alternative to euthanasia. We review the TNR literature in light of current debate over whether or not there should be further TNR trials in Australia. We revisit the problems arising from stray cats living in association with human habitation and estimate how many stray cats would have to be processed through a scientifically-guided TNR program to avoid high euthanasia rates. We also identify 10 ethical and welfare challenges that have to be addressed: we consider the quality of life for stray cats, where they would live, whether the TNR process itself is stressful, whether TNR cats are vulnerable to injury, parasites and disease, can be medically treated, stray cats' body condition and diet, and their impacts on people, pet cats, and urban wildlife, especially endemic fauna. We conclude that TNR is unsuitable for Australia in almost all situations because it is unlikely to resolve problems caused by stray cats or meet ethical and welfare challenges. Targeted adoption, early-age desexing, community education initiatives and responsible pet ownership have greater promise to minimize euthanasia, reduce numbers rapidly, and address the identified issues.

ZMIZ1 Variants Cause a Syndromic Neurodevelopmental Disorder.

ZMIZ1 is a coactivator of several transcription factors, including p53, the androgen receptor, and NOTCH1. Here, we report 19 subjects with intellectual disability and developmental delay carrying variants in ZMIZ1. The associated features include growth failure, feeding difficulties, microcephaly, facial dysmorphism, and various other congenital malformations. Of these 19, 14 unrelated subjects carried de novo heterozygous single-nucleotide variants (SNVs) or single-base insertions/deletions, 3 siblings harbored a heterozygous single-base insertion, and 2 subjects had a balanced translocation disrupting ZMIZ1 or involving a regulatory region of ZMIZ1. In total, we identified 13 point mutations that affect key protein regions, including a SUMO acceptor site, a central disordered alanine-rich motif, a proline-rich domain, and a transactivation domain. All identified variants were absent from all available exome and genome databases. In vitro, ZMIZ1 showed impaired coactivation of the androgen receptor. In vivo, overexpression of ZMIZ1 mutant alleles in developing mouse brains using in utero electroporation resulted in abnormal pyramidal neuron morphology, polarization, and positioning, underscoring the importance of ZMIZ1 in neural development and supporting mutations in ZMIZ1 as the cause of a rare neurodevelopmental syndrome.

Attitudes and Practices of Australian Veterinary Professionals and Students towards Early Age Desexing of Cats.

Surgical desexing of cats is typically carried out after six months of age (Mature Age Desexing, MAD); between 4⁻6 months (Traditional Age Desexing, TAD); or before four months (Early Age Desexing, EAD). We complemented existing surveys of veterinarians' acceptance of EAD with online and face-to-face surveys, to ascertain the preferred desexing ages for cats and rationale of 957 Australian veterinarians, veterinary nurses, veterinary science students, and veterinary nursing students. A complementary survey of 299 veterinary practice websites across Australia documented any information provided about desexing cats. The most common reason for preferred desexing ages was reducing stray cat populations (30%); 78% of these respondents chose ages aligning with EAD. Vet nurses and nursing students were more conservative than vets or vet students, preferring to desex cats >4 months. Perceived anaesthetic risk was a major motivation, especially for nurses ≤5 years' experience. Across 299 urban practices in Australian capital cities, 55% of surveyed websites provided no information about desexing cats or listed desexing without explaining why it was necessary, or when to perform it. Increasingly, Australian legislatures mandate desexing of cats by three months of age, so the practices of some current/future veterinary professionals do not match changing legislation.

Hair Follicle Dermal Cells Support Expansion of Murine and Human Embryonic and Induced Pluripotent Stem Cells and Promote Haematopoiesis in Mouse Cultures.

In the hair follicle, the dermal papilla (DP) and dermal sheath (DS) support and maintain proliferation and differentiation of the epithelial stem cells that produce the hair fibre. In view of their regulatory properties, in this study, we investigated the interaction between hair follicle dermal cells (DP and DS) and embryonic stem cells (ESCs); induced pluripotent stem cells (iPSCs); and haematopoietic stem cells. We found that coculture of follicular dermal cells with ESCs or iPSCs supported their prolonged maintenance in an apparently undifferentiated state as established by differentiation assays, immunocytochemistry, and RT-PCR for markers of undifferentiated ESCs. We further showed that cytokines that are involved in ESC support are also expressed by cultured follicle dermal cells, providing a possible explanation for maintenance of ES cell stemness in cocultures. The same cytokines were expressed within follicles in situ in a pattern more consistent with a role in follicle growth activities than stem cell maintenance. Finally, we show that cultured mouse follicle dermal cells provide good stromal support for haematopoiesis in an established coculture model. Human follicular dermal cells represent an accessible and readily propagated source of feeder cells for pluripotent and haematopoietic cells and have potential for use in clinical applications.

Missense Variants in RHOBTB2 Cause a Developmental and Epileptic Encephalopathy in Humans, and Altered Levels Cause Neurological Defects in Drosophila.

Although the role of typical Rho GTPases and other Rho-linked proteins in synaptic plasticity and cognitive function and dysfunction is widely acknowledged, the role of atypical Rho GTPases (such as RHOBTB2) in neurodevelopment has barely been characterized. We have now identified de novo missense variants clustering in the BTB-domain-encoding region of RHOBTB2 in ten individuals with a similar phenotype, including early-onset epilepsy, severe intellectual disability, postnatal microcephaly, and movement disorders. Three of the variants were recurrent. Upon transfection of HEK293 cells, we found that mutant RHOBTB2 was more abundant than the wild-type, most likely because of impaired degradation in the proteasome. Similarly, elevated amounts of the Drosophila ortholog RhoBTB in vivo were associated with seizure susceptibility and severe locomotor defects. Knockdown of RhoBTB in the Drosophila dendritic arborization neurons resulted in a decreased number of dendrites, thus suggesting a role of RhoBTB in dendritic development. We have established missense variants in the BTB-domain-encoding region of RHOBTB2 as causative for a developmental and epileptic encephalopathy and have elucidated the role of atypical Rho GTPase RhoBTB in Drosophila neurological function and possibly dendrite development.

Using Free Adoptions to Reduce Crowding and Euthanasia at Cat Shelters: An Australian Case Study.

Many healthy adult cats are euthanised annually in shelters, and novel approaches are required to reduce euthanasia rates. Waiving adoption fees is one such approach. However, concerns that less responsible owners will be attracted to free events persist among welfare groups. We evaluated evidence for differences in cat fate, health, and adherence to husbandry legislation via a case-study of a free adoption-drive for cats ≥1 year at a Western Australian shelter. Post-adoption outcomes were compared between free adopters and a control group of normal-fee adopters. The free adoption-drive rehomed 137 cats, increasing average weekly adoptions by 533%. First-time adopters were a significantly larger portion of the free cohort, as a result of mixed-media promotions. Both adopter groups selected cats of similar age; sex and pelage. Post-adoption, both groups retained >90% cats, reporting near identical incidences of medical and behavioural problems. Adopters did not differ in legislative compliance regarding fitting collars, registering cats, or allowing cats to roam. The shelter reported satisfaction with the adoption-drive, because in addition to relieving crowding of healthy adults, adoption of full-fee kittens increased 381%. Overall, we found no evidence for adverse outcomes associated with free adoptions. Shelters should not be dissuaded from occasional free adoption-drives during overflow periods.

Projections of the number of Australians with disability aged 65 and over eligible for the National Disability Insurance Scheme: 2017-2026.

OBJECTIVE: To develop projections of the size of the Australian population aged 65 years and over eligible for disability support through the National Disability Insurance Scheme (NDIS) for the decade following its introduction, to support planning and costing of the scheme. METHODS: We estimate disability and mortality transition probabilities and develop projections of the NDIS-eligible, ageing population from 2017 to 2026. RESULTS: An estimated 8000 men and 10 200 women aged 65 years and over will be eligible for support through the NDIS in 2017 (the scheme's first full year), increasing to 48 800 men and 56 900 women in 2026. CONCLUSIONS: Growth in the NDIS-eligible, ageing population has implications for relative budget allocations between the NDIS and the aged-care system, and projections of the size of this population are useful for calculating the overall cost of the NDIS.

Evidence for a Role of Prolactin in Mediating Effects of Photoperiod during the Dry Period.

Photoperiod manipulation during the lactation cycle alters milk yield, with long days (LDPP) increasing yield in lactation and short days (SDPP) in the dry period improving subsequent yield. Circulating prolactin (PRL) is directly related to day length, with LDPP increasing and SDPP decreasing PRL, respectively. Two blocks of 24 multiparous Holstein cows were used during two consecutive years to test the hypothesis that the mammary response to SDPP is the result of decreased concentrations of PRL in the circulation relative to LDPP. Cows were randomly assigned to one of three treatment groups during the dry period: SDPP, LDPP, or SDPP+PRL. Cows were returned to ambient photoperiod at calving and milk yield and DMI recorded for 120 d and 42 d, respectively. Mammary biopsies were obtained to determine rates of [³H]-thymidine incorporation into DNA in vitro. Treatment of SDPP cows with PRL caused a rapid increase in systemic PRL that reached concentrations similar to cows under LDPP. The periparturient PRL surge was similar for LDPP and SDPP+PRL cows, but those groups had greater surge concentrations versus SDPP. Cows exposed to SDPP produced more milk than LDPP cows, and there was a trend for SDPP+PRL cows to produce more milk than LDPP cows. Milk production was inversely related to the periparturient PRL surge. There was a trend for a treatment effect on mammary cell proliferation with greater proliferation in mammary tissue of SDPP cows relative to LDPP or SDPP+PRL on day -20 relative to parturition. Replacement of PRL to cows on SDPP when dry resulted in milk yield intermediate to cows on SDPP or LDPP, supporting the concept of a link between dry period PRL and yield.

NBAS mutations cause a multisystem disorder involving bone, connective tissue, liver, immune system, and retina.

We report two unrelated patients with a multisystem disease involving liver, eye, immune system, connective tissue, and bone, caused by biallelic mutations in the neuroblastoma amplified sequence (NBAS) gene. Both presented as infants with recurrent episodes triggered by fever with vomiting, dehydration, and elevated transaminases. They had frequent infections, hypogammaglobulinemia, reduced natural killer cells, and the Pelger-Huët anomaly of their granulocytes. Their facial features were similar with a pointed chin and proptosis; loose skin and reduced subcutaneous fat gave them a progeroid appearance. Skeletal features included short stature, slender bones, epiphyseal dysplasia with multiple phalangeal pseudo-epiphyses, and small C1-C2 vertebrae causing cervical instability and myelopathy. Retinal dystrophy and optic atrophy were present in one patient. NBAS is a component of the synthaxin-18 complex and is involved in nonsense-mediated mRNA decay control. Putative loss-of-function mutations in NBAS are already known to cause disease in humans. A specific founder mutation has been associated with short stature, optic nerve atrophy and Pelger-Huët anomaly of granulocytes (SOPH) in the Siberian Yakut population. A more recent report associates NBAS mutations with recurrent acute liver failure in infancy in a group of patients of European descent. Our observations indicate that the phenotypic spectrum of NBAS deficiency is wider than previously known and includes skeletal, hepatic, metabolic, and immunologic aspects. Early recognition of the skeletal phenotype is important for preventive management of cervical instability.

The effect of manual and mechanical stimulation on oxytocin release and milking characteristics in Holstein cows milked 3 times daily.

Prestimulation administered to a cow before attachment of the milking unit has historically been performed manually. Development of innovative milking technology has allowed manual stimulation to be replaced by mechanical forms of stimulation. Holstein cows (n=30) were enrolled in a crossover design to determine the effect of manual stimulation (forestripping and drying) and high-vibration pulsation on oxytocin profiles, milk yield, milk flow rates, incidence of delayed milk ejection causing bimodal milk flow curves, and residual milk in Holstein cows milked 3 times daily (3×). All cows were subjected to all treatments. Cows received manual (forestripping and drying) or mechanical (high-vibration pulsation) stimulation along with lag times of 0, 30, or 90 s for 21 consecutive milkings. Forestripping involved the manual removal of 2 streams of milk from each teat and drying of the teats. High-vibration pulsation involved increasing the pulsation cycles from 60 to 300 pulses/min and reducing the vacuum in the pulsation chamber to 20 kPa. The 5 treatments were (1) immediate attachment of the milking machine under normal pulsation (T0); (2) dip plus forestrip and drying with 30-s lag time (FD30); (3) dip plus forestrip and drying with 90-s lag time (FD90); (4) high-vibration pulsation for 30 s (HV30); and (5) high-vibration pulsation for 90 s (HV90). Milk yield per milking averaged 14.0 kg and was significantly different among treatments; however, the maximum difference detected among treatments was 0.8 kg/milking. Milking unit on-time, which represents the time when the milking unit is under normal pulsation and harvesting milk (excluding the high-vibration pulsation time of 30 or 90 s), was shortest (245 s) for cows subjected to 90 s of high-vibration pulsation (HV90) and ranged from 256 to 261 s for all other treatments. Milk harvest may have begun during high-vibration pulsation; however, only 0.13 and 0.32 kg of milk was harvested during high-vibration pulsation for HV30 and HV90, respectively. The incidence of bimodal milk curves was lowest for FD90 (7%) and highest for T0 (21%). The somatic cell count was <72×10(3) cells/mL for all treatments. Residual milk obtained by giving 10 IU of oxytocin in the jugular vein followed by 2 min of milking unit attachment represented 12 to 14% of the total milk and did not differ among treatments. Endogenous oxytocin profiles peaked between 12.4 and 18.3 pg/mL for all treatments, and the peak occurred sooner in manually stimulated cows; however, we detected no difference in oxytocin concentration beyond 2 min after milking unit attachment. High-vibration pulsation elicited a similar oxytocin release when taking the start time of stimulation from manual stimulation or high vibration into consideration. Forestripping for visual observation of milk combined with the use of high-vibration stimulation may reduce variation in the milking routine. A predetermined lag time with minimal variation may be achieved via the time spent in high-vibration stimulation instead of a lag period dictated by milking personnel.

TM4SF20 ancestral deletion and susceptibility to a pediatric disorder of early language delay and cerebral white matter hyperintensities.

White matter hyperintensities (WMHs) of the brain are important markers of aging and small-vessel disease. WMHs are rare in healthy children and, when observed, often occur with comorbid neuroinflammatory or vasculitic processes. Here, we describe a complex 4 kb deletion in 2q36.3 that segregates with early childhood communication disorders and WMH in 15 unrelated families predominantly from Southeast Asia. The premature brain aging phenotype with punctate and multifocal WMHs was observed in ~70% of young carrier parents who underwent brain MRI. The complex deletion removes the penultimate exon 3 of TM4SF20, a gene encoding a transmembrane protein of unknown function. Minigene analysis showed that the resultant net loss of an exon introduces a premature stop codon, which, in turn, leads to the generation of a stable protein that fails to target to the plasma membrane and accumulates in the cytoplasm. Finally, we report this deletion to be enriched in individuals of Vietnamese Kinh descent, with an allele frequency of about 1%, embedded in an ancestral haplotype. Our data point to a constellation of early language delay and WMH phenotypes, driven by a likely toxic mechanism of TM4SF20 truncation, and highlight the importance of understanding and managing population-specific low-frequency pathogenic alleles.

Human hair follicle dermal sheath and papilla cells support keratinocyte growth in monolayer coculture.

Traditional skin grafting techniques are effective but limited methods of skin replacement. Autologous transplantation of rapidly cultured keratinocytes is successful for epidermal regeneration, but the current gold-standard technique requires mouse fibroblast feeders and serum-rich media, with serum-free systems and dermal fibroblast (DF) feeders performing relatively poorly. Here, we investigated the capacity of human hair follicle dermal cells to act as alternative supports for keratinocyte growth. Dermal papilla (DP) dermal sheath (DS), DF and 3T3 cells were used as inactivated feeder cells for human keratinocyte coculture. Under conditions favouring dermal cells, proliferation of keratinocytes in the presence of either DS or DP cells was significantly enhanced compared with DF cells, at levels comparable to keratinocytes cultured under gold-standard conditions. Secreted protein acidic and rich in cysteine (SPARC) expression increased DS and DP cells relative to DFs; however, further experiments did not demonstrate a role in keratinocyte support.

The impact of social contagion on non-suicidal self-injury: a review of the literature.

In this review, we explore social contagion as an understudied risk factor for non-suicidal self-injury (NSSI) among adolescents and young adults, populations with a high prevalence of NSSI. We review empirical studies reporting data on prevalence and risk factors that, through social contagion, may influence the transmission of NSSI. Findings in this literature are consistent with social modeling/learning of NSSI increasing risk of initial engagement in NSSI among individuals with certain individual and/or psychiatric characteristics. Preliminary research suggests iatrogenic effects of social contagion of NSSI through primary prevention are not likely. Thus, social contagion factors may warrant considerable empirical attention. Intervention efforts may be enhanced, and social contagion reduced, by implementation of psychoeducation and awareness about NSSI in schools, colleges, and treatment programs.