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Avian pathogenic Escherichia coli (APEC) is the causative agent of avian colibacillosis and causes localized and/or systemic infections in poultry. The presence of various virulence genes (VGs) may be a useful marker for the detection of APEC directly from fecal samples. The objectives of this study were to evaluate and compare 3 different DNA extraction methods from cloacal swabs and fecal samples of broiler chickens and determine if APEC can be detected directly from feces. The DNA extraction methods were assessed by measuring DNA yield and purity, absence of DNA shearing, 16S ribosomal DNA amplification, and reproducibility. Repeated bead beating plus column (RBB+C) was the preferred extraction method, as it yielded an adequate amount of quality DNA for PCR directly from feces. The DNA extracted from feces, with RBB+C method and DNA extracted from E. coli isolates of organs and feces, taken from 23 broiler chickens (10 healthy, 9 with colibacillosis, and 4 unhealthy with other infections), were screened with a pentaplex-PCR for the prevalence of APEC-associated VGs: iroN, ompT, iutA, iss , and hlyF. There was a statistically significant correlation between the presence of the 5 VGs in E. coli cultured from the cloaca, fecal, and organs samples from chicken affected with colibacillosis. However, screening extracted DNA from the feces for the selected VGs was not an effective diagnostic tool to detect APEC as all of the VGs were detected in the extracted fecal DNA from all chickens. [ABSTRACT FROM AUTHOR]

Failure of lamellar energy metabolism may contribute to the pathophysiology of equine laminitis. Tissue microdialysis has the potential to dynamically monitor lamellar energy balance over time. The objectives of this study were to develop a minimally invasive lamellar microdialysis technique and use it to measure normal lamellar energy metabolite concentrations over 24 h. Microdialysis probes were placed (through the white line) into either the lamellar dermis (LAM) (n = 6) or the sublamellar dermis (SUBLAM) (n = 6) and perfused continuously over a 24 h study period. Probes were placed in the skin dermis (SKIN) for simultaneous comparison to LAM (n = 6). Samples were collected every 2 h and analysed for glucose, lactate, pyruvate, urea and glycerol concentrations. LAM was further compared with SUBLAM by simultaneous placement and sampling in four feet from two horses over 4 h. Horses were monitored for lameness, and either clinically evaluated for 1 month after probe removal (n = 4) or subjected to histological evaluation of the probe site (n = 10). There were no deleterious clinical effects of probe placement and the histological response was mild. Sample fluid recovery and metabolite concentrations were stable for 24 h. Glucose was lower (and lactate:glucose ratio higher) in LAM compared with SUBLAM and SKIN (P< 0.05). Pyruvate was lower in SUBLAM than SKIN and urea was lower in LAM than SKIN (P < 0.05). These differences suggest lower perfusion and increased glucose consumption in LAM compared with SUBLAM and SKIN. In conclusion, lamellar tissue microdialysis was well tolerated and may be useful for determining the contribution of energy failure in laminitis pathogenesis. [ABSTRACT FROM AUTHOR]

Heat treatment serves as a viable strategy to effectively mitigate the intense corrosion of biodegradable WE43 alloys. However, limited comprehension of the passivation mechanisms underlying heat treatment and the dilemma to quantitatively examine the evolution of hydrogen gas in vivo introduce uncertainties in designing heat treatments for developing clinically applicable WE43. This work aims to advance this knowledge by applying cutting-edge atom probe tomography to provide atomic-scale insights into the passivation roles of rare earth (RE)-rich β 1 (Mg 3 (Y, Nd)) and β' (Mg 12 NdY) nanophases induced by T6 heat treatment at 250 °C, and employing machine learning-based image analysis techniques to quantitatively unveil WE43's in vivo gas evolution during a 12-week implantation. It was found that nanosized β 1 and β' phases can effectively improve WE43's corrosion resistance by inducing an accelerated passivation effect on the surface and confining the distribution of hydrogen ions in the matrix. Female rats presented slightly higher corrosion rates than male rats in weeks 1 and 4 but lower hydrogen gas volumes in vivo, while male rats possessed a superior ability to metabolise hydrogen gas in vivo. Notably, latent gas evolution against the corrosion rates was found which peaked at week 4 and subsided at week 12 despite the gradually decreased corrosion rates from week 1 to 12. This study offers insights for engineering heat treatments to develop clinically applicable WE43 with acceptable corrosion rates and in vivo gas generation at various implantation stages. STATEMENT OF SIGNIFICANCE: The study aimed to reveal the role of β 1 and β' nanophases on the good corrosion resistance of WE43. The influence of these nanophases on WE43's corrosion performance has not been totally understood. Similarly, the understanding of hydrogen gas evolution as it relates to the magnesium implant's corrosion rate lacks clarity. Atom probe tomography (APT) indicates β 1 and β' nanophases trap hydrogen, removing H 2 from the lattice and disabling its catalytic role in Mg oxidation. Machine learning-aided analyses of computed tomography (CT) scan images indicate latent gas evolution, contradicting the monotonic in vivo H 2 evolution that is widely accepted., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

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.

The calf's gastrointestinal tract (GIT) microbiome undergoes rapid shifts during early post-natal life, which can directly affect calf performance. The objectives of this study were to characterise and compare differences in the establishment and succession of GIT microbiota, GIT morphological changes, and the growth of dairy calves from birth until weaned. Forty-four newborn Holstein-Friesian calves were randomly selected and assigned to Treatment (TRT) and Control (CON) groups. The TRT group calves received a once-daily dose of a direct-fed microbial (DFM) liquid product containing Lacticaseibacillus paracasei , Lentilactobacillus buchneri , and Lacticaseibacillus casei , all formerly known as Lactobacillus . Fresh faecal samples were manually taken from the rectum of all calves, and gross necropsy was performed on the forestomachs and gastrointestinal tracts. Bacterial DNA was extracted from frozen faecal samples for 16S rRNA gene amplicon sequencing. Calves in the TRT group had greater live weights ( p = 0.02) at weaning compared with calves in the CON group (mean = 69.18 kg, SD = 13.37 kg). The average daily live weight gain (ADG) and total feed intake were similar between the two groups. Calves in the TRT group had greater duodenum, abomasum, and reticulum weights ( p = 0.05). Rumen and intestinal development ( p < 0.05) and faecal microbial diversity ( p < 0.05) were more pronounced in the TRT group. The relative abundances of eight genera differed ( p < 0.001) between the groups. Supplementing calves with the LAB-based DFM increased live weight at weaning and had a more pronounced effect on the development of rumen and the gastrointestinal tract and on microbiota diversity and evenness. Future work is needed to better understand the potential association of LAB-DFM products on gut mucosa-associated microbiota.

Sudden and unexpected death (SUD) is a common reason for animals to undergo post-mortem examination. There is limited literature examining the causes of SUD in cats and dogs, and no research specific to Australia. The purpose of this study was to investigate the epidemiology and pathology of SUD in cats and dogs in a multicentric study across Australia. Retrospective post-mortem reports of SUD in cats and dogs were obtained from four veterinary schools in Australia distributed across four states. The frequency of SUD between institutes ranged from 2.1% to 6.5%. Dogs composed the majority of the study population (76%), and males outnumbered females, particularly in the feline subpopulation. After necropsy, 37% of SUD remained cause unknown, the largest category in both cats and dogs. When cause was identified, cardiovascular disease was most common in both species, followed by gastrointestinal disease in dogs, and trauma in cats. In dogs, multinomial logistic regression identified age as a risk factor significantly associated with the four largest categories of SUD. This study identified causes of SUD in Australian cats and dogs, including novel causes not previously reported. Further, this study revealed a higher rate of unsolved SUD in Australia than can be found in the literature from other countries.

A zinc-nutrient element alloy (Zn-1.0Cu-0.5Ca) was developed into subcuticular absorbable staples (SAS) as a robust alternative to the commercially available poly(l-lactide-co-glycolide) (PLGA) SAS for the first time. The fixation properties of the Zn SAS were measured via pull-out tests and in-situ lap-shear pull-out test comparatively against the PLGA SAS. The Zn SAS exhibited fixation force of 18.9±0.2 N, which was over three times higher than that of PLGA SAS (5.5±0.1 N). The Zn SAS was used to close incision wounds in a SD rat model for biodegradability and biocompatibility characterisation at 1, 4 and 12 weeks. The Zn SAS showed uniform degradation behaviour after in vivo implantation at the average rate of 198±54, 112±28, and 70±24 μm/y after 1, 4, and 12 weeks, which reduced the fixation force to 16.8±1.1 N, 15.4±0.9 N, 12.7±0.7 N, respectively. These findings showed the potential of the Zn SAS for the closure of heavy loading and slowing healing tissues. The Zn SAS enabled successful closure and healing of the incision wound, similar to the PLGA staples. However, the slow long-term degradation rate of the Zn SAS may lead to unnecessary implant retention. In addition, the alloy SAS resulted in higher local foreign body responses due to their stiffness. Reducing the implant cross-section profile and applying low stiffness and a corrosion-accelerating coating are suggested as possible approaches to reduce post-service implant retention and improve the biocompatibility of the Zn SAS. STATEMENT OF SIGNIFICANCE: This work reports the fabrication of the first metallic subcuticular absorbable staples (SAS) made from ZnCuCa alloy for skin wound closure applications. The Zn-based SAS were characterised in vitro and in vivo (SD rat model) for biodegradability, fixation properties, biocompatibility and inflammatory responses, which were compared against the commercially available PLGA-based SAS. The Zn-based SAS provided a secure attachment of the full-thickness wounds on SD rats and allowed successful healing during the 12-week service period. In addition, the in vitro results showed that the Zn-based SAS provided more than three times higher fixation strength than the commercial PLGA, indicating the potential of the Zn-based SAS for load-bearing wound closure application., (Copyright © 2023. Published by Elsevier Ltd.)

Populations of the iconic Australian koala are under constant decline. Their deaths are associated with rapid and extensive urbanization and the fragmentation of habitat areas. Using citizen science data on reported koala mortalities, we quantified the association between anthropogenic and environmental factors and the two leading causes of koala deaths, dog attacks and vehicle collisions. We achieved this objective using a case-control study design to compare the odds of exposure to a given risk factor for cases (a given cause of death) with the odds of exposure to a given risk factor for controls (all other causes of death). Koala deaths due to dog attacks were positively associated with registered dog population density and negatively associated with lot density whereas koala deaths due to vehicle collisions were positively associated with road density (road length per square kilometer) and negatively associated with human population density and distance to primary and secondary roads. The results of this research can be used to develop strategies to mitigate the risk of deaths due to dog attacks, for example by conducting educational awareness programs, promoting registration of dogs and discouraging free roaming of dogs. In a similar manner, in high-risk areas for vehicle collisions, over- or underpasses can be built to facilitate safe movement of koalas for road crossings or speed limits could be introduced to reduce the likelihood of premature koala deaths due to vehicle collisions., (© 2023. Springer Nature Limited.)

Crofton weed ( Ageratina adenophora ) is a global and highly invasive weed, with ingestion causing severe respiratory disease in horses, leading to irreversible and untreatable pulmonary fibrosis and oedema. While reports of equine pneumotoxicity remain common in Australia and New Zealand, equine pneumotoxicity may be underdiagnosed in other countries where Crofton weed is endemic but poorly differentiated. The pathogenesis of Crofton weed toxicity following ingestion has been well described in a number of different animal models, including rodents, rabbits, and goats. However, induced toxicity is organ-selective across different animal species, and these vastly differ from the pathogenesis described in horses, both clinically and after experimental exposure. Sources of variation may include species-specific susceptibility to different toxins present in the plant, different mechanistic processes of toxicity, and species differences in toxin biotransformation and bioactivation across different organs. Considering disease severity and Crofton weed's invasiveness globally, assessing published toxicological and exposure data is necessary to advance research, identify specific toxins for horses, and possible prophylactic and therapeutic strategies. This review presents an overview of the available literature on equine toxicity, parallels between toxicity in horses and other animal species, and important aspects to be included in the future research agenda.

This study investigated the role of cattle immune responses in the pathogenesis of buffalo fly ( Haematobia irritans exigua ) (BF) lesions. Brangus steers phenotyped for lesion development were divided into three groups: high lesion susceptibility (HL), low lesion susceptibility (LL) and no lesions (NL), based on lesion severity scores. Each steer was injected intradermally with different concentrations of BF, Onchocerca gibsoni (Og), and Musca domestica (Md) antigens. At 1 h post-injection, wheal areas at BF injection sites were found to be significantly larger in HL than NL cattle, but there were no significant differences ( p < 0.05) found between either the HL or NL cattle and LL cattle. At 24, 48, and 72 h post-injection, the skinfold thickness response to both BF and Md antigens was significantly greater in the HL group than the NL group. However, skin thickness was significantly greater for the BF antigens than the Md antigens ( p < 0.05). There were no significant differences found between the LL and NL animals in response to the BF antigens at any time, and no significant differences were determined between any of the lesion groups in response to the Og antigens. Histological examination of skin sections taken from the BF antigen injection sites in HL cattle at 72 h post-injection revealed necrosis of the epidermis and superficial dermis, along with severe eosinophilic inflammation. This study suggests that differences in the hypersensitivity to BF antigens underlie differences amongst the cattle in their susceptibility to the development of BF lesions, and breeding for immune-related biomarkers may assist in selecting more BF lesion-resistant cattle.

Liver disease in beef cattle has a significant global economic impact on feedlot and abattoir industries due to reduced animal performance, carcase yield, and carcase quality. This study aimed to create a post-mortem data collection tool which could be deployed at chain speed on an abattoir floor, as well as to evaluate pathological findings in both normal and condemned livers from an Australian beef cattle population. The first 1006 livers were used to formulate a user-friendly, high-throughput liver grading tool for use in an abattoir setting and to evaluate the histological features of common liver abnormalities. Subsequently, over 11,000 livers from a Southeast Queensland abattoir were analyzed. The most observed defects in condemned livers were liver abscessation, fibrosis, adhesions, and liver fluke, with histological features similar to previous reports. Bacterial culture was performed in 29 cases of liver abscessation, revealing a different balance of flora to that reported internationally. This study has developed an easy to use, efficient data collection tool that enables rapid, highly detailed assessment of large numbers of beef cattle livers at slaughter. This tool will allow thorough investigation into the effect of liver disease on beef production, in both industry and research contexts., Competing Interests: Declaration of Competing Interest None. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results., (Copyright © 2023. Published by Elsevier Ltd.)

Haematobia irritans exigua , commonly known as buffalo fly, is the major hematophagous ectoparasite of north Australian cattle herds. Lesions associated with buffalo fly infestation are generally alopecic, hyperkeratotic, or scab encrusted wounds with variable hemorrhagic ulceration. Buffalo flies can transmit a filarial nematode, Stephanofilaria sp., which has been implicated in the pathogenesis of buffalo fly lesions, but Stephanofilaria infection has not been detected in all lesions suggesting that other causal factors may be involved. This study characterized the pathology of buffalo fly lesions to identify the role of Stephanofilaria in lesion development, as well as to identify other potential agents. Lesion biopsies were collected from north and south Queensland and tested for the presence of Stephanofilaria by qPCR. Each lesion was scored grossly (0-4) for hemorrhage, ulceration, exudation, and alopecia. Lesions were also scored microscopically (0-4) for epidermal and dermal damage and inflammatory characters. Stephanofilaria infection was detected in 31% of lesion biopsies. Grossly, Stephanofilaria -infected lesions had significantly larger lesion area and higher scores for alopecia and hyperkeratosis than lesions where no nematodes were found ( P < 0.05). Histologically, epidermal, dermal, and adnexal damage was significantly higher in Stephanofilaria infected lesions than lesions without nematodes. Eosinophils, macrophages, and lymphocytes were significantly more abundant in Stephanofilaria positive lesions as compared to negative lesions. This study also noted bacterial infection with colonies of coccoid bacteria, observed in skin sections from 19 lesions. Grossly, lesions with bacterial infection had significantly higher ulceration scores compared to Stephanofilaria positive lesions, and histologically epidermal disruption was significantly greater in bacteria-infected lesions. We found no evidence of bacteria or Stephanofilaria infection in 49% of the lesions assessed and tissue damage patterns and eosinophilic inflammation suggested hypersensitivity to buffalo fly feeding as a possible cause of these lesions. These findings suggest that although the presence of Stephanofilaria infection may increase the severity of lesion pathology, it is not essential for lesion development. These outcomes also suggest a potential role of bacteria and hypersensitivity in pathogenesis of some lesion. A better understanding of buffalo fly lesion etiology will contribute to the optimal treatment and control programmes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Naseem, Allavena, Raza, Constantinoiu, McGowan, Turni, Kamran, Tabor and James.)

Despite the increased entry of women into the veterinary profession over the past several decades, women remain substantially underrepresented in senior leadership positions. This may include editorial positions at veterinary sciences journals. This study examines the gender distribution of editorships of 143 journals from the Thomson Reuters Web of Knowledge Journal Citation Reports category of veterinary science. The gender analysis was performed by impact factor, editorial board role, country and publisher. Females were underrepresented in the group of managing editors (32.2% females vs 67.2% males), editors (34.5% females vs 65.1% males) and others (33.3% females vs. 65.4% males). The journal impact factor did not have a significant impact on the proportion of males versus females on the editorial board. The median publisher had 27.5% editorships belonging to women. North America Europe and Oceania showed the greatest representation of women on editorial boards. Our findings provide the first measure of gender distribution on editorial boards in the veterinary sciences and may stimulate a discussion on the current situation of women in academic positions and, in particular, on editorial boards. Further research should investigate the underlying causes contributing to this aspect of gender inequality and identify possible strategies to encourage a greater participation of women to editorial boards., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Two sibling 12-week-old DNA-sexed female African grey parrots ( Psittacus erithacus ) were presented for progressive whole-body tremors, proprioceptive deficits, and an inability to stand unassisted. A third bird in the clutch (DNA-sexed as a male) exhibited no clinical signs. Physical examination of the affected birds revealed ataxia, inability to stand without assistance, and a reliance on their beaks to assist with their mobility. Hematologic and biochemical analyses were normal, as were radiographic images of both birds. Cerebellar disease of unknown origin was diagnosed, and the birds were euthanized. Postmortem examinations of the brains of both parrots revealed marked reduction in cerebellar size and poor folia formation. Microscopic review of the cerebellums demonstrated decreased density of the granular layer and thinning of the molecular layer with poorly organized and differentiated Purkinje fibers, consistent with a diagnosis of cerebellar hypoplasia. There are limited clinical reports and experimental studies examining cerebellar disease in birds. Conditions described have included cerebellar hypoplasia, cerebellar abiotrophy, and cerebellar dysplasia. Although these terms are used interchangeably due to similar clinical signs, histopathology is needed to differentiate between the different disease conditions. This case describes cerebellar hypoplasia that suggested a developmental etiology in 2 African grey parrots.

Commercial greyhound racing is legal in Australia but controversial due to concerns around animal welfare. To make evidence-based recommendations of animal welfare standards, a comprehensive analysis of available data on race events, animal health, injuries and fatalities is required. We undertook a review of publicly available data and reports published by official greyhound racing bodies for the purpose of determining how morbidity and mortality events associated with dog training and racing could be benchmarked. 6 years of available data from stewards' reports, quarterly and annual reports were descriptively analyzed from New South Wales, Victoria and Queensland. Whole-of-life tracking for individual dogs was sparse. Although stewards' reports were published in all three states, the availability of aggregated quarterly and annual reports varied. When available these provided additional information such as injury incidents standardized per thousand starts. In Queensland, quarterly and annual reports provided an overview of greyhound mortality and morbidity rates. In contrast with Victoria, quarterly reports were unavailable and only annual reports were published, meaning quarterly trends could not be determined. Therefore, injuries categorized by severity that were routinely included in quarterly reports in Queensland and New South Wales were unavailable in Victoria. Our findings demonstrate that data recording and reporting practices must be standardized to accurately evaluate whether animal welfare standards are being met in the Australian greyhound racing industry. Our recommendation is to have national standardized reporting of injuries and deaths, and a publicly available database for whole-of-life tracking for individual racing greyhounds., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chang, Descovich, Henning and Allavena.)

The objective of this study was to evaluate critically the nature and prevalence of histological pulmonary lesions in dogs and cats that had died or were euthanized because of tick paralysis. A retrospective and prospective case study of 11 cats and 23 dogs was carried out. Retrospective cases were gathered from the Veterinary Laboratory Services database at The University of Queensland (UQ). Prospective cases were provided by Veterinary Specialist Services and UQ VETs Small Animal Hospital. Lung and other tissue samples were collected for histopathological analysis. All tick intoxicated animals demonstrated evidence of pulmonary parenchymal changes: alveolar oedema, interstitial and alveolar congestion and alveolar fibrin exudation. Eleven of 23 (48%) dogs exhibited mild to severe bronchopneumonia. A lower rate (18%) of bronchopneumonia was found in cats, with one case of aspiration pneumonia. A novel pulmonary histological grading scheme was developed to evaluate the correlation between clinical presentation and histopathological changes. Novel extrapulmonary lesions in cats included hepatic necrosis and acute renal tubular necrosis attributed to hypoxia. We concluded that both dogs and cats with high clinical grade tick paralysis are extremely likely to have pulmonary pathology. High-protein oedema and fibrin exudation are predicted to be present in most cases of canine and feline tick paralysis., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Objective: To compare two commercial formulations of alfaxalone for immersion anaesthesia in laboratory zebrafish., Study Design: Prospective, blinded, randomized study., Animals: A total of 20 adult Danio rerio (Tuebingen strain)., Methods: Zebrafish were divided into two groups of 10 (five female, five male) and placed in individual immersion baths containing 10 mg L -1 of unpreserved alfaxalone (group 1) or preserved alfaxalone (group 2). Anaesthetists blinded to treatment used a composite score scale (CSS) (range 0-12) to assess fish every 30 seconds until induction of anaesthesia. Anaesthetic induction occurred when equilibrium and response to stimulus were lost. Fish were then placed in a clean water bath and scored every 60 seconds. Recovery from anaesthesia was defined as a CSS of ≤ 1. Time variables recorded were anaesthetic induction time (AIT), anaesthetic recovery time (ART) and total procedure time (TPT). Fish were observed for evidence of roupgross external pathology during the procedure. Following anaesthesia, four fish from each group were randomly chosen and euthanized for gill histopathology analysis immediately after recovery criteria were met. Data are presented as mean ± standard deviation. An independent t test was used to compare the difference in average anaesthetic time variables between groups (α = 0.05)., Results: There were no statistical differences between groups in reported variables. TPT, AIT and ART were 10.2 ± 1.2, 1.9 ± 0.9 and 8.3 ± 1.2 minutes for group 1 and 10.8 ± 2.9, 2.4 ± 1.2 and 8.4 ± 2.7 minutes for group 2. No gross external pathology was evident, and no fish died during the experimental period. Histopathology showed normal gill pathology and no difference between the groups., Conclusions and Clinical Relevance: Immersion anaesthesia using 10 mg L -1 of either formulation of alfaxalone resulted in anaesthesia of similar quality and duration., (Copyright © 2022 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Buffalo flies (Haematobia irritans exigua ) are hematophagous ectoparasites of cattle causing production and welfare impacts in northern Australian herds. Skin lesions associated with buffalo fly infestation and Stephanofilaria nematode infection are manifested as focal dermatitis or ulcerated areas, most commonly on the medial canthus of the eye, along the lateral and ventral neck, and on the abdomen of cattle. For closely related horn flies ( Haematobia irritans irritans ), Staphylococcus aureus has been suggested as a contributing factor in the development of lesions. To investigate the potential role of bacterial infection in the pathogenesis of buffalo fly lesions, swabs were taken from lesions and normal skin, and bacteria were also isolated from surface washings of buffalo flies and surface-sterilized homogenized flies. Bacterial identification was conducted by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and strain typing by repetitive sequence-based PCR (rep-PCR) and DNA sequencing to determine species similarity and virulence factors. Of 50 bacterial isolates collected from lesions, 38 were identified as Staphylococcus agnetis and 12 as Staphylococcus hyicus, whereas four isolates from normal skin were S. hyicus and one was Mammaliicoccus sciuri. Of the Staphylococcus isolates isolated from buffalo flies, five were identified as S. agnetis and three as S. hyicus . Fifty percent of the buffalo fly isolates had rep-PCR genotypic patterns identical to those of the lesion isolates. Genome sequencing of 16 S. agnetis and four S. hyicus isolates revealed closely similar virulence factor profiles, with all isolates possessing exfoliative toxin A and C genes. The findings from this study suggest the involvement of S. agnetis and S. hyicus in buffalo fly lesion pathogenesis. This should be taken into account in the development of effective treatment and control strategies for lesions. IMPORTANCE Skin lesions in cattle associated with feeding by Haematobia fly species are a significant welfare issue in Australia, North and South America, and Europe. The development of these lesions has been attributed to a number of causal factors, but the exact etiology and pathogenesis were unclear. This study characterized Staphylococcus agnetis and Staphylococcus hyicus strains from cattle skin lesions and in vector flies and demonstrated their role in the pathogenesis of these lesions. These findings will aid the development of targeted and more effective treatment and control strategies for lesions associated with fly infestation in cattle.

Inter-pathologist variation is widely recognized across human and veterinary pathology and is often compounded by missing animal or clinical information on pathology submission forms. Variation in pathologist threshold levels of resident inflammatory cells in the tissue of interest can further decrease inter-pathologist agreement. This study applied a predictive modeling tool to bladder histology slides that were assessed by four pathologists: first without animal and clinical information, then with this information, and finally using the predictive tool. All three assessments were performed twice, using digital whole-slide images (WSI) and then glass slides. Results showed marked variation in pathologists' interpretation of bladder slides, with kappa agreement values of 7-37% without any animal or clinical information, 23-37% with animal signalment and history, and 31-42% when our predictive tool was applied, for digital WSI and glass slides. The concurrence of test pathologists to the reference diagnosis was 60% overall. This study provides a starting point for the use of predictive modeling in standardizing pathologist agreement in veterinary pathology. It also highlights the importance of high-quality whole-slide imaging to limit the effect of digitization on inter-pathologist agreement and the benefit of continued standardization of tissue assessment in veterinary pathology.

Buffalo flies (Haematobia irritans exigua) are ectoparasites of major animal health and production concern in north Australian beef herds. Skin lesions associated with buffalo fly infestation, cause hide damage and welfare issues and are manifested as dermatitis or ulcerated areas found most commonly near the medial canthus of the eye, along the lateral and ventral neck and on the abdomen. Buffalo flies can transmit a nematode, Stephanofilaria sp., which has been considered the main aetiological agent for buffalo fly lesions, but the role of nematodes in the development of the lesions has not been defined. To investigate the geographical distribution of Stephanofilaria, swabs were taken from the surface of lesions and buffalo flies were netted from the backs of beef cattle from 20 properties located in northern, central and southern Queensland. Swabs and buffalo flies were then tested for the presence of Stephanofilaria by qPCR. In northern and central Queensland, all properties except one, tested positive for the presence of Stephanofilaria in either buffalo flies or swabs, or in both. The infection rate varied amongst sites ranging from 0% to 100% in lesions and 0-34% in female buffalo flies. No nematodes were found in male buffalo flies. In contrast, none of the 66 lesion swabs or 1220 buffalo flies collected from southern Queensland tested positive for Stephanofilaria infection despite the frequent occurrence of lesions in the herds from which samples were collected. These findings suggest that infection with Stephanofilaria, although frequently detected, is not essential for the development of buffalo fly lesions and other factors may contribute to the initiation of lesions. This study also confirmed the potential for using surface swabs as a quicker and less invasive means of sampling lesions than dermal biopsies when testing for the presence of Stephanofilaria by qPCR, but further studies will be required to estimate the sensitivity of this technique. Understanding the pathogenesis of buffalo fly lesions will aid the development of optimal treatment and control strategies., (Copyright © 2022. Published by Elsevier B.V.)

This case series describes polycystic kidney disease in 3 (2 male, 1 female) 2-month-old, juvenile rainbow lorikeets ( Trichoglossus moluccanus ). The lorikeets diagnosed with polycystic kidney disease were the progeny of full sibling parents that were being intentionally line bred for the purpose of establishing a rainbow lorikeet with the blue color mutation. Clinically the juvenile lorikeets were presented with clinical signs of lethargy, dehydration, regurgitation, anorexia, polyuria, and pelvic limb paresis. Multiple abnormalities were identified on the complete blood count and plasma biochemistry panel, including a normocytic normochromic nonregenerative anemia, hyperuricemia, hyperphosphatemia, hypercalcemia, and azotemia. Severe renal dysfunction was diagnosed in all birds on the basis of clinical presentation, physical examination, and complete blood count and plasma biochemistry results. Radiographically marked renomegaly was noted in one of the cases. Although intensive critical care and supportive therapy was provided, 1 lorikeet died, and the remaining 2 were euthanatized because of client financial constraints and a rapid deterioration of their clinical condition associated with severe renal dysfunction. Postmortem pathology results found that all birds had marked renomegaly, visceral gout, and polycystic kidney disease. Because of the age of the birds and the line breeding within this group of lorikeets, the disease was believed to be inherited. Polycystic kidney disease should be considered as a possible differential diagnosis in juvenile psittacine birds with a history of line breeding when presented with severe renal dysfunction. From the current case series, polycystic kidney disease appears to carry a grave prognosis in juvenile rainbow lorikeets.

A magnesium alloy containing essential, non-toxic, biodegradable elements such as Ca and Zn has been fabricated using a novel twin-roll casting process (TRC). Microstructure, mechanical properties, in vivo corrosion and biocompatibility have been assessed and compared to the properties of the rare earth (RE) element containing WE43 alloy. TRC Mg-0.5 wt% Zn- 0.5 wt% Ca exhibited fine grains with an average grain size ranging from 70 to 150 μm. Mechanical properties of a TRC Mg-0.5Zn-0.5Ca alloy showed an ultimate tensile strength of 220 MPa and ductility of 9.3%. The TRC Mg-0.5Zn-0.5Ca alloy showed a degradation rate of 0.51 ± 0.07 mm/y similar to that of the WE43 alloy (0.47 ± 0.09 mm/y) in the rat model after 1 week of implantation. By week 4 the biodegradation rates of both alloys studied were lowered and stabilized with fewer gas pockets around the implant. The histological analysis shows that both WE43 and TRC Mg-0.5Zn-0.5Ca alloy triggered comparable tissue healing responses at respective times of implantation. The presence of more organized scarring tissue around the TRC Mg-0.5Zn-0.5Ca alloys suggests that the biodegradation of the RE-free alloy may be more conducive to the tissue proliferation and remodelling process., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

Background: Stephanofilaria spp. nematodes are associated with cutaneous lesions in cattle and other livestock and mammalian wildlife species. In Australia, Haematobia irritans exigua, commonly known as buffalo fly (BF) transmits a well-described but presently unnamed species of Stephanofilaria , which has been speculatively implicated in the aetiology of BF lesions. The sensitivity of current techniques for detecting Stephanofilaria spp. in skin lesions and vector species is low, and there is no genomic sequence for any member of the genus Stephanofilaria currently available in sequence databases., Methods: To develop molecular assays for the detection of the Australian Stephanofilaria sp., skin biopsies were collected from freshly slaughtered cattle with typical lesions near the medial canthus. Adult nematodes and microfilariae were isolated from the biopsies using a saline recovery technique. The nematodes were morphologically identified as Stephanofilaria sp. by scanning electron microscopy. DNA was extracted and the internal transcribed spacer 2 (ITS2) region of rDNA, and the cytochrome c oxidase subunit 1 ( cox1 ) region of mtDNA was amplified and sequenced. Stephanofilaria sp. specific polymerase chain reaction (PCR) and qPCR assays (SYBR Green ® and TaqMan™) were developed and optimised from the novel ITS2 sequence obtained. The specificity of each assay was confirmed by testing against nematode species Onchocerca gibsoni and Dirofilaria immitis , as well as host (bovine) and BF DNA., Results: Scanning electron microscopy of the anterior and posterior ends of isolated nematodes confirmed Stephanofilaria sp. A phylogenetic analysis of the cox1 sequence demonstrated that this species is most closely related to Thelazia callipaeda , a parasitic nematode that is a common cause of thelaziasis (or eyeworm infestation) in humans, dogs, and cats. Both conventional and qPCR assays specifically amplified DNA from Stephanofilaria sp. Conventional PCR, TaqMan™, and SYBR Green ® assays were shown to detect 1 ng, 1 pg, and 100 fg of Stephanofilaria DNA, respectively. Both qPCR assays detected DNA from single Stephanofilaria microfilaria., Conclusion: Molecular diagnostic assays developed in this study showed high specificity and sensitivity for Stephanofilaria sp. DNA. The availability of an accurate and sensitive PCR assay for Stephanofilaria will assist in determining its role in the pathogenesis of cattle skin lesions, as well as in understanding its epidemiological dynamics. This assay may also have application for use in epidemiological studies with other species of Stephanofilaria , most particularly closely related S. stilesi , but this will require confirmation.

The koala, Phascolarctos cinereus , is an iconic Australian wildlife species facing a rapid decline in South-East Queensland (SEQLD). For conservation planning, the ability to estimate the size of koala populations is crucial. Systematic surveys are the most common approach to estimate koala populations but because of their cost they are often restricted to small geographic areas and are conducted infrequently. Public interest and participation in the collection of koala sighting data is increasing in popularity, but such data are generally not used for population estimation. We modeled monthly sightings of koalas reported by members of the public from 1997 to 2013 in SEQLD by developing a self-exciting spatio-temporal point process model. This allowed us to account for characteristics that are associated with koala presence (which vary over both space and time) while accounting for detection bias in the koala sighting process and addressing spatial clustering of observations. The density of koalas varied spatially due to the heterogeneous nature of koala habitat in SEQLD, with a mean density of 0.0019 koalas per km 2 over the study period. The percentage of land areas with very low densities (0-0.0005 koalas per km 2 ) remained similar throughout the study period representing, on average, 66% of the total study area. The approach described in this paper provides a useful starting point to allow greater use to be made of incidental koala sighting data. We propose that the model presented here could be used to combine systematic koala survey data (which is spatially restricted, but more precise) with koala sighting data (which is incidental and often biased by nature, but often collected over large geographical areas). Our approach could also be adopted for modeling the density of other wildlife species where data is collected in the same manner., Competing Interests: None declared., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Background: We describe intratumoral injection of a slow-release emulsion of killed mycobacteria (complete Freund's adjuvant (CFA)) in three preclinical species and in human cancer patients., Methods: Efficacy and safety were tested in mammary tumors in mice, in mastocytomas in mice and dogs, and in equine melanomas. In mice, survival, tumor growth, and tumor infiltration by six immune cell subsets (by flow cytometry) were investigated and analyzed using Cox proportional hazards, a random slopes model, and a full factorial model, respectively. Tumor growth and histology were investigated in dogs and horses, as well as survival and tumor immunohistochemistry in dogs. Tumor biopsies were taken from human cancer patients on day 5 (all patients) and day 28 (some patients) of treatment and analyzed by histology. CT scans are provided from one patient., Results: Significantly extended survival was observed in mouse P815 and 4T1 tumor models. Complete tumor regressions were observed in all three non-human species (6/186 (3%) of mouse mastocytomas; 3/14 (21%) of canine mastocytomas and 2/11 (18%) of equine melanomas). Evidence of systemic immune responses (regression of non-injected metastases) was also observed. Analysis of immune cells infiltrating mastocytoma tumors in mice showed that early neutrophil infiltration was predictive of treatment benefit. Analysis of the site of mastocytoma regression in dogs weeks or months after treatment demonstrated increased B and T cell infiltrates. Thus, activation of the innate immune system alone may be sufficient for regression of some injected tumors, followed by activation of the acquired immune system which can mediate regression of non-injected metastases. Finally, we report on the use of CFA in 12 human cancer patients. Treatment was well tolerated. CT scans showing tumor regression in a patient with late-stage renal cancer are provided., Conclusion: Our data demonstrate that intratumoral injection of CFA has major antitumor effects in a proportion of treated animals and is safe for use in human cancer patients. Further trials in human cancer patients are therefore warranted. Our novel treatment provides a simple and inexpensive cancer immunotherapy, immediately applicable to a wide range of solid tumors, and is suitable to patients in developing countries and advanced care settings., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)