A RETROSPECTIVE ANALYSIS OF MORBIDITY AND MORTALITY IN THE CAPTIVE LEADBEATER'S POSSUM (GYMNOBELIDEUS LEADBEATERI) POPULATION FROM 1970 TO 2021.

The Leadbeater's possum (Gymnobelideus leadbeateri) is a critically endangered nocturnal marsupial with a restricted range in the Central Highlands of Victoria, Australia. There are two genetically distinct populations divided by location: highland and lowland. Lowland possums exist in one remnant swamp forest and entered captivity in 2012 when ∼60 individuals remained. Today, with less than 20 lowland individuals remaining, any information that informs the yet-unsuccessful breeding program is critical. This study encompasses a retrospective analysis of the causes of mortality and significant histological lesions in captive highland and lowland individuals across seven institutions internationally from 1970 to 2021. During this time, 245 possums lived in captivity. Postmortem records exist for 99 animals, including 349 histopathology diagnoses from 80 reports and 264 gross necropsy diagnoses from 78 reports. Diagnoses were assigned into two categories based on the importance to the individual (causing death or morbidity to a single animal [n = 194]), or importance to the wider population (causing death or morbidity to more than one animal or was related to reproduction [n = 155]). Individual animals had multiple diagnoses, which were tallied as individual data points. Renal disease was diagnosed 57 times; the most common finding was chronic nephropathy (43/57). Cardiovascular disease was diagnosed 33 times; atherosclerosis associated with obesity was common (n = 10/33). Both categories suggest causal association with captive husbandry but elicit no comment on the lack of success of the breeding program. Reproductive disease was diagnosed 36 times in 24 animals (14 females and 10 males). In females, 11 cases of uterine inflammation and associated clinical signs were associated with ascending infection or neoplasia. Of the seven lowland male possums with mortality data, five were infertile (azoospermia or testicular atrophy). More investigation into the reproductive health of this population is indicated to understand the lack of success in the current breeding program.

Establishing key performance indicators for inflammatory bowel disease in the UK.

Background and aims: Healthcare quality improvement (QI) is the systematic process to continuously improve the quality of care and outcomes for patients. The landmark Inflammatory Bowel Disease (IBD) UK National Audits provided a means to measure the variation in care, highlighting the need to define the standards of excellence in IBD care. Through a consensus approach, we aimed to establish key performance indicators (KPIs), providing reliable benchmarks for IBD care delivery in UK. Methods: KPIs that measure critical aspects of a patient journey within an IBD service were identified though stakeholder meetings. A two-stage Delphi consensus was then conducted. The first involved a multidisciplinary team of IBD clinicians and patients to refine definitions and methodology. The second stage assessed feasibility and utility of the proposed QI process by surveying gastroenterology services across UK. Results: First, the four proposed KPIs were refined and included time from primary care referral to diagnosis in secondary care, time to treatment recommendation following a diagnosis, appropriate use of steroids and advanced therapies prescreening and assessment. Second, the Delphi consensus reported >85% agreement on the feasibility of local adoption of the QI process and >75% agreement on the utility of benchmarking of the KPIs. Conclusions: Through a structured approach, we propose quantifiable KPIs for benchmarking to improve and reduce the individual variation in IBD care across the UK.

Severe pericardial effusion in a cat with peritoneopericardial diaphragmatic hernia and incarcerated hepatic sarcoma.

Case summary: A 4-year-old female neutered domestic longhair cat was presented at a referral hospital for dyspnoea with a history of suspected pleural effusion. Thoracic ultrasonography demonstrated a large-volume pericardial effusion causing cardiac tamponade and a cystic mass within the pericardium. CT revealed a peritoneopericardial diaphragmatic hernia (PPDH) caused by a defect of the ventral diaphragm. Herniated contents consisted of the right lateral and caudate liver lobes, and an associated cystic hepatic mass. Ventral midline coeliotomy was performed for herniorrhaphy and partial pericardiectomy, together with lobectomy of the incarcerated liver mass. Histopathology and immunohistochemistry diagnosed a poorly differentiated hepatic sarcoma with inflammation and remodelling in the adjacent incarcerated liver parenchyma. The patient developed metastatic sarcoma 2 months after surgery and was euthanased as a result. Relevance and novel information: Pericardial effusion causing cardiac tamponade is a previously unreported sequelae to PPDH in cats. Reports on the presence of malignancy in incarcerated liver are scarce and the location is not typical for a sarcoma in this species.

Canine mast cell tumours part I: Clinical and survival outcomes.

BACKGROUND: Dogs have a species-specific susceptibility for developing mast cell tumours (MCTs). Mutations in the KIT proto-oncogene (KIT) are known to contribute to the neoplastic biology of mast cells. In dogs, the most common KIT mutation is an internal tandem duplication (ITD) in exon 11 which has been considered a useful prognostic supplement to traditional histopathological tumour grading. OBJECTIVE: The aim of this retrospective study was to explore the importance of KIT exon 11 ITD mutation status and known clinical and pathological indices in predicting prognosis in a cohort of Australian dogs diagnosed with MCT. METHODS: Clinical parameters, survival data, and KIT mutation status were collected and assessed for 220 dogs with cutaneous or subcutaneous MCT (n = 189 and n = 31, respectively). RESULTS: In at least one of the multivariable models, tumour grade (cutaneous Kiupel low or high grade) or tumour subcutaneous location, multiple concurrent MCTs, metastasis at the time of surgery, and senior age were statistically significant in predicting the outcome (MCT-related death and/or second MCT diagnosis) at 6- or 12-month post-tumour excision. KIT exon 11 ITD mutation status was not a significant predictor in any of the final multivariable models and was strongly correlated with high histological grade (p < 0.001). CONCLUSION: In this sample of dogs, tumour histological grading remained the single most powerful prognostic indicator for MCT outcome. However, concurrent evaluation of multiple prognostically significant parameters provides information of potential value to inform therapeutic management for each patient.

Detection of Breinlia sp. (Nematoda) in the Leadbeater's possum (Gymnobelideus leadbeateri).

The Leadbeater's possum (Gymnobelideus leadbeateri) is a critically endangered marsupial in south-eastern Australia. Among other conservation efforts, free-ranging animals in the two remaining geographically separate populations (highland and lowland) have been extensively studied; however, little is known about their health and mortality. Although some wild populations are frequently monitored, cadavers are rarely recovered for post mortem examination. In June 2019, a recently deceased, wild, adult male lowland Leadbeater's possum was collected from a nest box and a comprehensive post mortem examination was conducted. Microfilariae of a filarioid nematode were observed in testes, liver, lung and skin samples in tissue impression smears and upon histopathological examination. No gross or histological changes were seen associated with the parasites, except for a focal area of tissue damage in the skin, suggesting that the possum is a natural host. Using a PCR-coupled sequencing method the filarioid was identified as a species of Breinlia. Species of Breinlia occur in other Australian marsupials and rodents.

Disseminated protozoal infection in a wild feathertail glider (Acrobates pygmaeus) in Australia.

This is the first report of a disseminated protozoal infection in a wild feathertail glider (Acrobates pygmaeus) from south-eastern Australia. The glider was found dead in poor body condition. Histologically, large numbers of zoites were seen predominantly in macrophages in the liver, spleen and lung, with protozoal cysts present in the liver. Molecular and phylogenetic analyses inferred that the protozoan parasite belongs to the family Sarcocystidae and is closely related to previously identified apicomplexans found in yellow-bellied gliders (Petaurus australis) in Australia and southern mouse opossums (Thylamys elegans) in Chile.

Service-Learning through conference-based, interdisciplinary workshops on Age-Friendly design.

For three consecutive years, the Age-Friendly Design Committee (AFDC) of the Academy for Gerontology in Higher Education (AGHE) used AGHE's annual meeting as a platform to conduct community-based service-learning workshops focusing on age-friendly design. These workshops assembled local stakeholders, conference attendees from multiple disciplines, and landscape and architectural designers to discuss age-friendly design issues and solutions for local environments. Each workshop provided hands-on design experience and the opportunity for AGHE participants to contribute to conference host communities by using their gerontological expertise to translate knowledge into practice. Local stakeholders learned the value of gerontological input when considering design issues. We describe the process of incorporating service-learning into the conference experience through age-friendly design workshops and how these bring together students, faculty, and design professionals from different backgrounds and disciplines to address local age-friendly design issues.

SURVEILLANCE FOR CHLAMYDIA SPP. WITH MULTILOCUS SEQUENCE TYPING ANALYSIS IN WILD AND CAPTIVE BIRDS IN VICTORIA, AUSTRALIA.

Chlamydia psittaci typically infects birds and can cause outbreaks of avian chlamydiosis, but it also has the potential to cause zoonotic disease (psittacosis) in humans. To better understand the epidemiology of C. psittaci in Victoria, Australia, we conducted opportunistic sampling of more than 400 wild and captive birds presented to the Australian Wildlife Health Centre at Zoos Victoria's Healesville Sanctuary for veterinary care between December 2014 and December 2015. Samples were screened for the presence of chlamydial DNA using quantitative PCR, and positive samples were subjected to multilocus sequence typing analysis. The results showed a significantly higher prevalence of infection in captive birds (8%; 9/113) compared to wild birds (0.7%; 2/299). Multilocus sequence typing analysis revealed that C. psittaci sequence type 24 was detected in both wild and captive birds in the local region, while C. psittaci sequence type 27 was detected for the first time in an Australian avian host. The generally low prevalence of C. psittaci detection points to a generally low zoonotic risk to veterinary and support staff, although this risk may be higher when handling captive birds, where the prevalence of C. psittaci infection was almost 10-fold higher. Even with low rates of C. psittaci detection, appropriate hygiene and biosecurity practices are recommended due to the serious human health implications of infection with this pathogen.

A randomised vaccine field trial in Kenya demonstrates protection against wildebeest-associated malignant catarrhal fever in cattle.

Wildebeest-associated malignant catarrhal fever (WA-MCF), a fatal disease of cattle caused by alcelaphine herpesvirus 1 (AlHV-1), is one of the most important seasonal diseases of cattle in wildebeest endemic areas, with annual incidence reaching 10%. Here we report efficacy of over 80% for a vaccine based on the attenuated AlHV-1 C500 strain, in preventing fatal WA-MCF in cattle exposed to natural wildebeest challenge. The study was conducted at Kapiti Plains Ranch Ltd, south-east of Nairobi, Kenya. In 2016, 146 cattle were selected for a randomised placebo-controlled trial. Cattle were stratified according to breed and age and randomly assigned to groups given vaccine or culture medium mixed with Emulsigen®. Cattle received prime and boost inoculations one month apart and few adverse reactions (n = 4) were observed. Indirect ELISA demonstrated that all cattle in the vaccine group developed a serological response to AlHV-1. The study herd was grazed with wildebeest from one month after booster vaccination. Three cattle, two that received vaccine and one control, succumbed to conditions unrelated to WA-MCF before the study ended. Twenty-five cattle succumbed to WA-MCF; four of the remaining 71 cattle in the vaccine group (5.6%) and 21 of the remaining 72 control cattle (29.2%; χ2 = 13.6, df = 1, p < 0.001). All of the WA-MCF affected cattle were confirmed by PCR to be infected with AlHV-1 and in 23 cases exhibited histopathology typical of WA-MCF. Vaccine efficacy was determined to be 80.6% (95% CI 46.5-93.0%). Hence, the AlHV-1 C500 vaccine is a safe and potentially effective novel method for controlling WA-MCF in cattle. The implementation of this vaccine may have significant impacts on marginalised cattle keeping communities.

DNA purification increases PCR-amplifiable DNA extracted from formalin-fixed, paraffin-embedded canine mast cell tumors for routine KIT mutation detection.

DNA amplification by PCR detects KIT exon 11 internal tandem duplications in canine mast cell tumors (MCTs). Tissue-specific inhibitors often contaminate DNA extracted from formalin-fixed, paraffin-embedded (FFPE) canine MCTs, blocking PCR amplification and, consequently, preventing mutation detection. We used a commercial kit to extract DNA from FFPE canine MCTs. Two independent PCR assays, each with one primer set, were used to amplify target genes (HPRT and KIT) directly after FFPE DNA extraction. PCR amplification failed with at least one primer set in 153 of 280 samples (54.6%, 95% CI: 48.8-60.5%). One or 2 DNA washing steps were required to remove PCR inhibitors in 130 of 280 (46.4%) and 23 of 280 (8.2%) of these cases, respectively. DNA concentration and quality (A260/A280 and A260/A230) either pre- or post-washing were not associated with ability of the samples to be amplified by PCR using both HPRT and KIT primer sets. Low-grade and subcutaneous MCTs were less likely to amplify directly after DNA extraction and without any washing steps compared to high-grade MCTs using KIT gene primers.

PREVALENCE OF COLUMBID HERPESVIRUS INFECTION IN FERAL PIGEONS FROM NEW SOUTH WALES AND VICTORIA, AUSTRALIA, WITH SPILLOVER INTO A WILD POWERFUL OWL (NINOX STRUENA).

Columbid herpesvirus-1 (CoHV-1) is widespread in feral pigeons in North America and Europe. We used a PCR assay to detect CoHV-1 DNA in oral and cloacal tissues and oral swabs from naturally infected pigeons. Fifty-three feral pigeons from five flocks in Australia (n=3 from south-central Victoria and n=2 from Sydney) were examined for CoHV-1 DNA. We detected CoHV-1 DNA in oral mucosa and cloacal mucosa, with higher concentrations in the oral mucosa. The sensitivity of testing oral swabs was the same as testing the tissue, indicating that testing of oral swabs from live birds is an effective means of screening flocks for CoHV-1 infection. Infection was found in all five of the flocks examined and the prevalence of infection ranged from 70% to100%. Most positive birds could be detected with a single-amplification PCR, but a nested amplification was required to detect others. Oral swabs from Australian native doves and pigeons (n=18) and the introduced Collared Dove (Streptopelia chinensis; n=2) were also tested by the nested PCR and all were negative for CoHV-1 DNA. We describe a fatal infection of CoHV-1 in a wild Powerful Owl (Ninox strenua) that was observed feeding on feral pigeons. This is the first known case of CoHV-1 causing death in a wild bird of prey in Australia. Our data suggest that CoHV-1 is widespread in feral pigeon flocks in Australia but we did not find it in native doves and pigeons. Spillover into native avian predator species may be occurring.

Salicylic acid treatment and expression of an RNA-dependent RNA polymerase 1 transgene inhibit lethal symptoms and meristem invasion during tobacco mosaic virus infection in Nicotiana benthamiana.

BACKGROUND: Host RNA-dependent RNA polymerases (RDRs) 1 and 6 contribute to antiviral RNA silencing in plants. RDR6 is constitutively expressed and was previously shown to limit invasion of Nicotiana benthamiana meristem tissue by potato virus X and thereby inhibit disease development. RDR1 is inducible by salicylic acid (SA) and several other phytohormones. But although it contributes to basal resistance to tobacco mosaic virus (TMV) it is dispensable for SA-induced resistance in inoculated leaves. The laboratory accession of N. benthamiana is a natural rdr1 mutant and highly susceptible to TMV. However, TMV-induced symptoms are ameliorated in transgenic plants expressing Medicago truncatula RDR1. RESULTS: In MtRDR1-transgenic N. benthamiana plants the spread of TMV expressing the green fluorescent protein (TMV.GFP) into upper, non-inoculated, leaves was not inhibited. However, in these plants exclusion of TMV.GFP from the apical meristem and adjacent stem tissue was greater than in control plants and this exclusion effect was enhanced by SA. TMV normally kills N. benthamiana plants but although MtRDR1-transgenic plants initially displayed virus-induced necrosis they subsequently recovered. Recovery from disease was markedly enhanced by SA treatment in MtRDR1-transgenic plants whereas in control plants SA delayed but did not prevent systemic necrosis and death. Following SA treatment of MtRDR1-transgenic plants, extractable RDR enzyme activity was increased and Western blot analysis of RDR extracts revealed a band cross-reacting with an antibody raised against MtRDR1. Expression of MtRDR1 in the transgenic N. benthamiana plants was driven by a constitutive 35S promoter derived from cauliflower mosaic virus, confirmed to be non-responsive to SA. This suggests that the effects of SA on MtRDR1 are exerted at a post-transcriptional level. CONCLUSIONS: MtRDR1 inhibits severe symptom development by limiting spread of virus into the growing tips of infected plants. Thus, RDR1 may act in a similar fashion to RDR6. MtRDR1 and SA acted additively to further promote recovery from disease symptoms in MtRDR1-transgenic plants. Thus it is possible that SA promotes MtRDR1 activity and/or stability through post-transcriptional effects.

Cutaneous T-cell lymphoma in two captive Tasmanian devils (Sarcophilus harrisii).

Two captive adult female Tasmanian devils (Sarcophilus harrisii) were investigated for pruritis and dermatitis. In both cases skin lesions consisted of multifocal, superficial patches of crusting, hyperkeratosis, and ulceration. Lesions started on the ventral surfaces of the animal but then appeared on the dorsum as the disease progressed. In both animals, a diagnosis of cutaneous T-cell lymphoma was made based on histologic appearance of skin biopsies using immunohistochemistry. Attempt at treatment with lomustine 20 mg p.o. once every 3 wk in one individual did not slow progression of the condition. As a result of their propensity for developing neoplastic conditions, the use of chemotherapeutic agents in Tasmanian devils warrants further investigation.

Benign gastric neuroendocrine tumors in three snow leopards (Panthera uncia).

Neuroendocrine tumors are relatively rare neoplasms arising from neuroendocrine cells that are distributed throughout the body and are predominant in the gastrointestinal tract. This report describes benign, well-differentiated gastric neuroendocrine tumors in three captive snow leopards (Panthera uncia). All tumors were well circumscribed, were within the gastric mucosa or submucosa, and had histologic and immunohistochemical features of neuroendocrine tumors. Histologic features included packeted cuboidal to columnar epithelial cells that were arranged in palisades or pseudorosettes and contained finely granular cellular cytoplasm with centrally placed, round nuclei. Cytoplasmic granules of neoplastic cells strongly expressed chromogranin A, variably expressed neuron-specific enolase, and did not express synaptophysin or gastrin. Each leopard died or was euthanatized for reasons unrelated to its tumor.

Non-West Nile virus-associated mortality in a population of American crows (Corvus brachyrhynchos): a gross and histopathologic study.

The American crow (Corvus brachyrhynchos) is a common urban and rural inhabitant of the Northeast and Midwest United States that is commonly infected with West Nile virus (WNV). The current study was initiated to determine non-WNV-associated causes of mortality in the American crow. All animals (40/40) tested negative for WNV infection via polymerase chain reaction and had no evidence of infection based on immunohistochemistry. Common gross necropsy findings included external trauma (6/40), hepatosplenomegaly (6/40), poxviral dermatitis (5/40), and pneumonia (3/40). Common histologic findings included endoparasitism (32/40), multifocal hepatic and splenic necrosis (7/40), pigment accumulation in the spleen (5/40), and disseminated bacterial infection (3/40). The most significant and debilitating diseases included fungal pneumonia and poxvirus-associated lesions. The present report increases the knowledge of diseases present in the American crow population.