Unveiling the Role of Endoplasmic Reticulum Stress Pathways in Canine Demodicosis.

Canine demodicosis is a prevalent skin disease caused by overpopulation of a commensal species of Demodex mite, yet its precise cause remains unknown. Research suggests that T-cell exhaustion, increased immunosuppressive cytokines, induction of regulatory T cells and increased expression of immune checkpoint inhibitors may contribute to its pathogenesis. This study aimed to gain a deeper understanding of the molecular changes occurring in canine demodicosis using mass spectrometry and pathway enrichment analysis. The results indicate that endoplasmic reticulum stress promotes canine demodicosis through regulation of three linked signalling pathways: eIF2, mTOR, and eIF4 and p70S6K. These pathways are involved in the modulation of Toll-like receptors, most notably TLR2, and have been shown to play a role in the pathogenesis of skin diseases in both dogs and humans. Moreover, these pathways are also implicated in the promotion of immunosuppressive M2 phenotype macrophages. Immunohistochemical analysis, utilising common markers of dendritic cells and macrophages, verified the presence of M2 macrophages in canine demodicosis. The proteomic analysis also identified immunological disease, organismal injury and abnormalities and inflammatory response as the most significant underlying diseases and disorders associated with canine demodicosis. This study demonstrates that Demodex mites, through ER stress, unfolded protein response and M2 macrophages contribute to an immunosuppressive microenvironment, thereby assisting in their proliferation.

Descriptive epidemiology of smothering in Australian commercial free-range layer hen farms.

Since the early 2000 s the practice of free-range egg production has increased in developed countries, partly driven by consumer perception that free-range housing is better for hen welfare. While poultry in free-range systems have more behavioural opportunities compared with poultry in caged systems, free-range systems are associated with greater frequencies of infectious disease, predation and 'smothering', a condition where birds pile on top of one another with death occurring due to suffocation. Although the frequency of smothering deaths in Australian free-range layer poultry is anecdotally high, there is a lack of empirical evidence quantifying smothering cause-specific mortality rates and identifying factors that place birds at higher risk of death from smothering. This was a prospective cohort study of poultry flocks managed by three commercial free-range layer organisations in Eastern Australia. Flocks were enrolled into the study from 1 January 2019 to 29 March 2021 and were followed until the end of lay or until the end of the study on 31 March 2022, whichever occurred first. Throughout the follow-up period flock managers provided production details for each flock and details of smothering events using custom-designed logbooks.A total of 84 flocks were enrolled in the study: 32 from Organisation 1, 35 from Organisation 2 and 17 from Organisation 3. The number of birds per flock ranged from 16,000 to 45,000. The total mortality rate was 1131 deaths per 10,000 bird-years. Smothering mortality rate across the three organisations was 183 (minimum 133, maximum 223) deaths per 10,000 bird-years at risk. Smothering accounted for around 16% (minimum 9%, maximum 22%) of all deaths.We identified no distinctive temporal pattern in daily smothering risk as a function of either the number of days since placement or calendar date. The locations of smothering events in sheds and in the outdoor range were not consistent, with relatively large numbers of smothering events occurring in specific locations for some sheds but not others. To the best of our knowledge, this study is the largest prospective study of smothering mortality in commercial free-range layer flocks conducted to date. Estimates of smothering incidence rate and how that varies within and between flocks and organisations over time provides a critically important benchmark for further investigations into this substantial area of productivity loss.

Early life events associated with first lactation reproductive performance in southwest Victorian pasture-based dairy herds.

This was a prospective cohort study to determine how events from birth until first calving affect reproductive performance in the first lactation in pasture-based dairy herds in Victoria, Australia. Events during the preweaning (0 to 84 days), weaning to first breeding (85 to 473 days) and first breeding to first calving periods (474 to 804 days) were recorded and their association with reproductive performance during the first lactation was quantified. Reproductive performance outcomes included the number of days from first mating start date to first service (MSD-S1) and the number of days from first mating start date to first conception (MSD-CON). Predictors for reproductive performance included: passive transfer status as a calf; season of birth; age and breed of the dam; breed; the presence or absence of dystocia at the time of the heifer's birth; active feeding of colostrum versus being left on the dam for colostrum intake; presence of twinning; the presence or absence of preweaning diarrhoea; preweaning, prepubertal and postpubertal average daily gain; the presence or absence of periparturient disease at first calving; age at first calving; body condition score at first calving; sex of the first-born calf; the presence or absence of a stillborn calf at the first calving and requirement of assistance at first calving. Two Cox proportional hazards regression models were developed: the first for early life event variables associated with MSD-S1 and the second for early life events associated with MSD-CON. Heifers born in autumn and winter had 2.89 (95% CI 1.50 to 5.59, P = 0.002) times and 1.97 (95% CI 1.12 to 3.44, P = 0.018) times the daily hazard of being inseminated compared with heifers born in spring, respectively. For the MSD-S1 analyses heifers that gave birth to a live calf had 1.64 (95% CI 1.14 to 2.36, P = 0.008) times the daily hazard of being inseminated compared with heifers that had a stillborn calf. Increases in weight gain during the first breeding to first calving period by 0.1 kg/day increments increased the daily hazard of first insemination by a factor of 1.10 (95% CI 1.00 to 1.22, P = 0.043). Heifers that experienced periparturient disease had a significantly lower hazard of conception per day compared with heifers that did not experience periparturient disease at the first calving (HR 0.67, 95% CI 0.50 to 0.91, P = 0.009). Increases in weight gain during the first breeding to first calving period by 0.1 kg/day increased the daily hazard of conception by a factor of 1.10 (95% CI 1.01 to 1.21, P = 0.038). We conclude that of all the growth periods assessed in this study, events that occurred during the first breeding to first calving period (474 to 804 days) had the greatest association with reproductive performance in the first lactation. There should be a focus on increasing growth rates during this period and reducing the risk of stillbirth and periparturient disease to improve reproductive performance in the subsequent mating period after calving.

Bodyweight and body condition scores of Australian British shorthaired cats, 2008-2017.

Obesity is the most common nutritional problem in companion animals today, and Australian British shorthair (BSH) cats have been shown to have a greater likelihood of being overweight relative to other cat breeds. The objectives of this research were to quantify bodyweight (BW) and body condition scores (BCSs) of BSH cats attending first opinion practice in Australia for the period 2008-2017 and to determine if: (1) being classified as overweight was associated with geographical location (urban versus rural and socio-economic index); and (2) BW recorded in the first 12 months of life was associated with length of life beyond 12 months. Electronic medical records from BSH cats were obtained from VetCompass Australia and used for BW and BCS analysis. Desexed males (n = 971) had the highest mean BW followed by entire males (n = 79), desexed females (n = 815), and entire females (n = 82). The desexed males, desexed females, and entire females had a mean BCS classified as overweight using a 1-to-9 point BCS scale. The entire male population was the only group with a mean BCS classified as ideal. No statistically significant association between BW and urban-rural status and no consistent trend between BW and socioeconomic deprivation was found. For cats with at least one BW measurement in the first 12 months of life that was ≤3.3 kg, the age when 20 percent of the group had died or were euthanised was 12.3 (95% CI 11.7 to 13.1) years. For cats with at least one BW measurement in the first 12 months of life that was ≥3.3 kg age, the age when 20 percent of the group had died or were euthanised was 6.6 (95% CI 5.2 to 6.6) years. This was a substantial clinical difference in survival. The study concluded that a large proportion of BSH cats attending first opinion veterinary clinics in Australia between 2008 and 2017 (48%) were classified as overweight. Cats less than 12 months of age that were greater than 3.3 kg had a shortened lifespan beyond 12 months of age compared with cats that were less than 3.3 kg.

Scenario tree modelling to inform surveillance design for maintaining freedom from Coxiella burnetii infection in Australian commercial dairy goat herds.

We used scenario tree methods to determine how different disease detection methods might be used to provide quantitative evidence that Australian dairy goat herds are free of coxiellosis. The aim of our proposed C. burnetii surveillance programme is to find evidence of the absence of antigen as well as evidence of an absence of an immune response to C. burnetii infection in individual dairy goat herds. We defined a C. burnetii infected dairy goat herd as a herd in which at least one doe was showing evidence of either active infection or past C. burnetii exposure using four candidate surveillance system components (SSCs): (1) testing of individual doe whole blood using the C. burnetii com1 PCR; (2) testing of individual doe whole blood using the C. burnetii ELISA; (3) testing bulk tank milk (BTM) using the com1 PCR and the C. burnetii ELISA; and (4) investigations of abortions and stillborn kids submitted to a diagnostic laboratory for testing. Of eight candidate surveillance strategies (combinations of the SSCs listed above) individual doe ELISAs every six months combined with monthly BTM PCR and ELISA testing returned the highest surveillance system sensitivity of 0.963 (95% probability interval [PI] 0.911-0.982) for the lowest cost, at AUD 28.94 (95% PI 28.38-30.59) over a 12-month period, for every one percent increase in surveillance system sensitivity. Assuming a probability of disease freedom of 0.10 at the start of the surveillance program and a probability of C. burnetii introduction per month of 0.01 we estimate that 95% confidence that C. burnetii was absent from a herd could be achieved after a single round of individual doe ELISAs followed by period of 6 consecutive monthly BTM PCR and ELISA tests. The results of this study show that selection of the most efficient combination of surveillance system components requires a good understanding of initial herd C. burnetii status and the probability of introduction of infection and how this may change over time. Scenario tree analyses results have provided insight into the key determinants of C. burnetii detection ability.

National participation levels in the 2017-2018 Australian thoroughbred racing season.

INTRODUCTION: The social licence of the Australian thoroughbred (TB) industry relies on the general public's perception of how they manage the animal in their care. METHOD: This study examines the horse, race and activity records for the 37,704 horses racing and training in Australia from 1 August 2017 to 31 July 2018. Three-quarters (75%, n = 28,184) of TBs started in one of the 180, 933 race starts that occurred during the 2017-2018 Australian racing season. RESULTS: Horses participating in the 2017-2018 Australian racing season had a median age of 4 years, with geldings more likely to be aged 5 years and older. The majority of the TB racehorse population were geldings (51%, n = 19,210), with 44% (n = 16,617) females and only 5% (n = 1877) entire males. Horses aged 2 years were three times more likely to not start in a race in that year compared with older horses. At the end of the 2017-2018 racing season, 34% of the population had an inactive status recorded. Horses aged 2 years (median two starts) and 3 years (median five starts) had fewer starts than older horses (median seven starts). Eighty-eight percent (n = 158,339) of race starts were over distances of 1700 metres or less. Race starts by horses aged 2 years (46%, 3264 of 7100) were more likely to have occurred at a metropolitan meeting compared with starts by older horses. CONCLUSION: This study provides a national overview of the racing and training activities and TBs participating in the 2017-2018 Australian racing season.

Validating the female fertility estimated breeding value in Australian commercial dairy herds.

We conducted a retrospective cohort study to validate the efficacy of the Australian multitrait fertility estimated breeding value (EBV). We did this by determining its associations with phenotypic measures of reproductive performance (i.e., submission rate, first service conception rate, and early calving). Our secondary aim was to report the associations between these reproductive outcomes and management and climate-related factors hypothesized to affect fertility. Our study population included 38 pasture-based dairy herds from the northern Victorian irrigation region in Australia. We collected records for 86,974 cows with 219,156 lactations and 438,578 mating events from the date on which managers started herd recording until December 2016, comprising both fertility-related data such as insemination records, calving dates, and pregnancy test results, and systems-related data such as production, herd size, and calving pattern. We also collected hourly data from 2004 to 2017 from the closest available weather station to account for climate-related factors (i.e., temperature humidity index; THI). Multilevel Cox proportional hazard models were used to analyze time-to-event outcomes (days to first service, days to cow calving following the planned herd calving start date), and multilevel logistic regression models for binomial outcomes (conception to first service) in the Holstein-Friesian and Jersey breeds. A 1-unit increase in daughter fertility EBV was associated with a 5.4 and 8.2% increase in the daily hazard of calving in the Holstein-Friesian and Jersey breeds respectively. These are relative increases (i.e., a Holstein-Friesian herd with a 60% 6-wk in-calf rate would see an improvement to 63.2% with a 1-unit increase in herd fertility EBV). Similar results were obtained for submission and conception rate. Associations between 120-d milk yield and reproductive outcome were complicated by interactions with 120-d protein percentage and calving age, depending on the breed and outcome. In general, we found that the reproductive performance of high milk-yielding animals deteriorated faster with age than low milk-yielding animals, and high protein percentage exacerbated the differences between low and high milk-yielding animals. Climate-related factors were also associated with fertility, with a 1-unit increase in maximum THI decreasing first service conception rate by 1.2% for Holstein-Friesians but having no statistically significant association in the Jersey breed. However, THI had a negative association in both breeds on the daily hazard of calving. Our study validates the efficacy of the daughter fertility EBV for improving herd reproductive performance and identifies significant associations between 120-d milk and protein yields and THI on the fertility of Australian dairy cows.

Health surveillance representative of koala (Phascolarctos cinereus) distribution in Victoria, Australia.

Health surveillance of wildlife populations is essential for conservation and reduction of the impacts of disease. Population declines and areas of overabundance of koalas (Phascolarctos cinereus) can disrupt the overall survival of the species as well as its habitat. This retrospective study was conducted to describe population distributions, identify areas which need increased surveillance and improve koala health surveillance methodology by Wildlife Health Victoria: Surveillance (WHV:S) at the Veterinary School of The University of Melbourne. Twelve years of Victorian koala observation data from the Atlas of Living Australia combined with surveillance data from WHV:S were used to create choropleth maps, using Quantum Geographic Information Systems of populations and surveillance events, visually representing hot spots. This data was further used to calculate health surveillance efforts between 2008 to the beginning of 2020. Analysis ranked postcodes throughout Victoria from low surveillance efforts to high, using standardised surveillance ratio's 95% confidence interval upper limits which were mapped using a colour gradient. This identified postcodes which need increased surveillance effort, corresponding to areas with high koala observations and low surveillance submissions. This analysis can guide surveillance for postcodes with koalas that were under-represented and inform improved methodology of future surveillance by WHV:S. The specific advice for improvements to WHV:S includes utilisation of citizen science and syndromic surveillance, website improvement, increasing community awareness and more. The limitations of this study were discussed.

A cross-sectional survey of risk factors for the presence of Coxiella burnetii in Australian commercial dairy goat farms.

The largest Australian farm-based outbreak of Q fever originated from a dairy goat herd. We surveyed commercial dairy goat farms across Australia by testing bulk tank milk (BTM) samples using a commercial indirect enzyme-linked immunosorbent assay and two quantitative polymerase chain reactions (PCRs). Of the 66 commercial dairy goat herds on record, managers from 61 herds were contacted and 49 provided BTM samples. Five of the surveyed herds were positive on at least one of the diagnostic tests, thus herd-level apparent prevalence was 10% (95% confidence interval [CI] 4 to 22). True prevalence was estimated to be 3% (95% credible interval: 0 to 18). Herd managers completed a questionnaire on herd management, biosecurity and hygiene practices and risk factors were investigated using multivariable logistic regression. Herds with >900 milking does (the upper quartile) were more likely to be Coxiella burnetii positive (odds ratio = 6.75; 95% CI 1.65 to 27.7) compared with farms with ≤900 milking does. The odds of BTM positivity increased by a factor of 2.53 (95% CI 1.51 to 4.22) for each order of magnitude increase in the number of goats per acre. C. burnetii was not detected in samples from the majority of the Australian dairy goat herds suggesting there is an opportunity to protect the industry and contain this disease with strengthened biosecurity practices. Intensification appeared associated with an increased risk of positivity. Further investigation is required to discriminate the practices associated with an increased risk of introduction to disease-free herds, from practices associated with maintenance of C. burnetii infection in infected dairy goat herds.

A machine-learning assisted review of the use of habit formation in medication adherence interventions for long-term conditions.

Adherence to medication in long-term conditions is around 50%. The key components of successful interventions to improve medication adherence remain unclear, particularly when examined over prolonged follow-up periods. Behaviour change theories are increasingly interested in the utility of habit formation for the maintenance of health behaviour change, but there is no documentation on how habit has been conceptualised in the medication adherence intervention literature, or what effect the key technique identified in habit formation theory (context dependent repetition) has in these studies. To examine this, a machine-learning assisted review was conducted. Searches of MEDLINE, EMBASE and PSYCInfo and the reference list of a comprehensive systematic review of medication adherence interventions yielded 5973 articles. Machine learning-assisted title and abstract screening identified 15 independent RCTs published between 1976 and 2021, including 18 intervention comparisons of interest. Key findings indicate that conceptualisations of habit in the medication adherence literature are varied and behaviour change technique coding identified only six studies which explicitly described using habit formation. Future work should aim to develop this evidence base, drawing on contemporary habit theory and with explicit demonstration of what techniques have been used to promote habit formation.

Factors associated with racing performance and career duration for Victorian-born Thoroughbreds.

The number of horses leaving the Australian Thoroughbred (TB) racing industry each year is of concern to animal welfare advocates, public and regulators. A horse's previous athletic performance is a significant driver of retirement from racing. Racehorse performance can be measured in terms of the total number of starts, duration of racing and prize money earned. This study investigated Australian racing records for the 2005 and 2010 Victorian TB foal crops to identify factors associated with total number of starts, racing career duration, prize money earned and age of last race start-up to the 10-year-old racing season. Racing Australia registered 4,577 TB horses born in Victoria in 2005 (n = 2,506) and 2010 (n = 2,071) that raced in Australia. Horses that started racing at 2-years of age had fewer race starts in their first racing season but an increased total number of starts, prize money and duration of racing. The median age of last start (LS) was five (Q1 4; Q3 7) years. Horses that had won a race, had a maximum handicap rating of 61 or above and those racing over distances of more than 2,400 m had an increased racing career duration and an age of last race start greater than 6-years of age. Horses participating in jumps races (n = 63) had the longest careers and older age of LS. These horses were more likely to have had a handicap rating over 80 and were just as likely to start their racing careers as 2-years-olds.

Participation of Victorian Thoroughbreds in the racing industry: a whole-of-population benchmark.

The proportion of Thoroughbred (TB) horses that commence training and eventually race, is an important industry-level indicator of how successful it is at producing horses suitable for racing. It is also of interest to the wider community and is central to the issue of whether there is overbreeding in the Australian TB industry. This study investigated the training and racing records for the 2005 and 2010 Victorian TB foal crops to determine the proportion of TBs that train and race, and the age of entering training and first race start. Subsets of the foal crop were also examined to determine if premier yearling sale cohorts would produce selection bias. The Australian Stud Book (ASB) registered 7662 TB horses born in Victoria in 2005 (n = 4116) and 2010 (n = 3546). Of these, 5614 (73%) entered training and 4868 (64%) started in at least one race. Fourteen percent (n = 1045) of the study cohort had their first race start as 2-year-olds and 35% (n = 2644) had their first start as 3-year-olds. A higher proportion of TB horses that attended premier yearling sales entered training (93%) and raced (84%). This study established a baseline for training and racing milestones, such as the proportion and age that horses commenced training, and age of first start using population-level data. It also established that the use of subsets of the population can introduce selection bias.

Persistent HIV-1 transcription in CD4+ T cells from ART-suppressed individuals can originate from biologically competent proviruses.

HIV-1 is able to persist in the face of potent antiretroviral therapy (ART). A number of strategies are being explored to allow ART-free viral remission or viral eradication. In order to gauge the progress of these strategies, assays with which to measure viral reservoir size and activity are needed. In a large percentage of aviremic individuals on suppressive ART, viral transcripts can be detected in peripheral blood CD4+ T cells. While this cell-associated RNA has been considered as a marker of viral reservoir activity, it is unclear whether cell-associated viral transcripts in aviremic individuals originate from biologically competent proviruses as opposed to being a product of abortive transcription from defective proviruses. We assessed whether cell-associated viral RNA in peripheral blood CD4+ T cells from aviremic individuals on ART originated from biologically competent proviruses. We demonstrate that cell-associated viral RNA transcripts were highly related to viral sequences obtained by ex vivo outgrowth. This relationship was also observed when viral transcription in the outgrowth cultures was limited to donor CD4+ T cells. Our study indicates that cell-associated viral RNA warrants further consideration as a viral reservoir surrogate in individuals on suppressive ART.

Bayesian latent class analysis when the reference test is imperfect.

Latent class analysis (LCA) has allowed epidemiologists to overcome the practical constraints faced by traditional diagnostic test evaluation methods, which require both a gold standard diagnostic test and ample numbers of appropriate reference samples. Over the past four decades, LCA methods have expanded to allow epidemiologists to evaluate diagnostic tests and estimate true prevalence using imperfect tests over a variety of complex data structures and scenarios, including during the emergence of novel infectious diseases. The objective of this review is to provide an overview of recent developments in LCA methods, as well as a practical guide to applying Bayesian LCA (BLCA) to the evaluation of diagnostic tests. Before conducting a BLCA, the suitability of BLCA for the pathogen of interest, the availability of appropriate samples, the number of diagnostic tests, and the structure of the data should be carefully considered. While formulating the model, the model's structure and specification of informative priors will affect the likelihood that useful inferences can be drawn. With the growing need for advanced analytical methods to evaluate diagnostic tests for newly emerging diseases, LCA is a promising field of research for both the veterinary and medical disciplines.

L'analyse à classes latentes a permis aux épidémiologistes de surmonter les problèmes concrets posés par les méthodes traditionnelles d'évaluation des essais de diagnostic, qui nécessitent à la fois un test de référence absolue (étalon ou gold standard) et un grand nombre d'échantillons de référence aux caractéristiques appropriées. Au cours des quatre dernières décennies, les méthodes d'analyse à classes latentes ont acquis de l'ampleur et permettent aux épidémiologistes d'évaluer les essais diagnostiques et d'estimer les taux de prévalence réelle tout en recourant à des tests supposés imparfaits, grâce à l'utilisation de données et de scénarios divers et complexes, y compris dans les situations d'émergence de nouvelles maladies infectieuses. Les auteurs font un tour d'horizon des dernières évolutions dans ce domaine et donnent des orientations pratiques concernant la manière d'utiliser l'analyse bayésienne à classes latentes pour évaluer les performances d'un test de diagnostic. Avant de conduire une telle analyse, il convient de déterminer avec soin si elle est adaptée à l'agent pathogène considéré et si les échantillons disponibles sont appropriés et en nombre suffisant ; il convient également de prendre en compte le nombre de tests de diagnostic à évaluer et la structure des données utilisées. Lors de la conception du modèle, sa structure et la définition préalable des données informatives vont affecter la probabilité que le modèle génère des inférences utiles. Face à la nécessité croissante de disposer de méthodes analytiques sophistiquées pour évaluer les tests de diagnostic utilisés pour les maladies émergentes nouvelles, les analyses à classes latentes offrent des perspectives prometteuses pour la recherche, aussi bien dans le domaine de la santé vétérinaire que de la médecine humaine.

El análisis de clases latentes ha servido a los epidemiólogos para superar las limitaciones prácticas que imponen los métodos tradicionales de evaluación de pruebas de diagnóstico, que requieren a la vez una prueba de diagnóstico que sirva de patrón de referencia perfecto y un gran número de muestras de referencia adecuadas. En los últimos cuatro decenios, los métodos de análisis de clases latentes se han ido ampliando hasta permitir a los epidemiólogos evaluar pruebas de diagnóstico y calcular la prevalencia real empleando pruebas imperfectas ante muy diversas estructuras de datos y situaciones complejas, incluida la aparición de nuevas enfermedades infecciosas. Los autores, tras presentar a grandes líneas los últimos adelantos en cuanto a métodos de análisis de clases latentes, ofrecen indicaciones prácticas para aplicar el análisis bayesiano de clases latentes a la evaluación de pruebas de diagnóstico. Antes de proceder a un análisis bayesiano de este tipo conviene estudiar con detenimiento la idoneidad del método para el patógeno en cuestión, la disponibilidad de muestras apropiadas, el número de pruebas de diagnóstico y la estructura de los datos. A la hora de formular el modelo, la estructura del propio modelo y la especificación de los elementos informativos previos influirán en la probabilidad de poder extraer conclusiones provechosas. Ante la creciente necesidad de disponer de métodos analíticos avanzados con los que evaluar pruebas de diagnóstico de nuevas enfermedades emergentes, el análisis de clases latentes abre un promisorio campo de investigación para las disciplinas veterinarias y médicas.

Diagnostic test validation studies when there is a perfect reference standard.

Before tools became available to consider diagnostic test validation studies where a 'gold-standard' is not available, new diagnostic tests were compared to a reference standard assumed to be highly accurate if not perfect. This paper reviews such 'traditional' situations with examples and methods of study design and analysis. Three situations are described, two where a perfect reference is available for either positive or negative animals, and one where the reference is perfect for both. Thus, here the authors review circumstances to be considered when validating a diagnostic test with a credible reference standard. An appropriate study design requires an unbiased selection of animals from the population to which a new test will be applied. Examples for calculating sample size and data analysis are provided. Finally, the authors discuss situations where it may be appropriate to include influential variables ('covariates') in a diagnostic test validation study..

Avant la mise au point d'outils permettant de concevoir des études de validation pour des tests candidats en l'absence d'un étalon de référence à l'exactitude parfaite (référence absolue ou gold standard), les tests à valider étaient comparés à un étalon de référence censé présenter un niveau élevé d'exactitude, à défaut d'être parfait. Les auteurs décrivent ces situations « classiques ¼ en donnant quelques exemples et en précisant les méthodes employées pour la conception et l'analyse de ces études. Ils décrivent trois situations : dans deux d'entre elles, un étalon de référence parfait existe, soit pour les animaux positifs, soit pour les animaux négatifs ; dans la troisième situation, l'étalon de référence est parfait pour les deux catégories. Ainsi, les circonstances prises en compte ici sont celles de la validation d'un test diagnostique au moyen d'un réactif de référence crédible. Une conception d'étude appropriée passe par une sélection non biaisée des individus composant l'échantillon au sein de la population animale à laquelle sera appliqué le test. Les auteurs donnent quelques exemples de calcul de la taille de l'échantillon et d'analyse des données. Enfin, ils examinent les situations où il peut être opportun d'inclure des variables influentes (« covariance ¼) dans l'étude de validation d'un test diagnostique.

Antes de que apareciesen herramientas que permiten juzgar los estudios de validación de pruebas de diagnóstico en ausencia de un modelo o patrón de calibración, las nuevas pruebas de diagnóstico eran comparadas con un patrón de referencia considerado muy exacto, cuando no perfecto. Los autores se refieren aquí a este tipo de situaciones "clásicas", exponiendo ejemplos y métodos de diseño de estudios y análisis de los datos obtenidos. Los autores describen tres tipos de situación: cuando se dispone de una referencia perfecta para animales positivos; cuando se dispone de una referencia perfecta para animales negativos; y cuando la referencia es perfecta para ambos casos. Para cada tipo de situación examinan los aspectos que hay que tener en cuenta al validar una prueba de diagnóstico con un patrón de referencia fiable. El diseño adecuado de un estudio exige una selección no sesgada de los animales a partir de una población a la que vaya a aplicarse la nueva prueba. Los autores ofrecen ejemplos del modo de calcular el tamaño muestral y analizar los datos de un estudio. Por último, examinan situaciones en las que pueda ser conveniente incluir en el estudio de validación uno o más factores que puedan influir (covariables).

Emerging and endemic zoonotic diseases: surveillance and diagnostics.

In this paper, the authors: (a) list methods used to diagnose zoonotic diseases in humans and animals; (b) identify between-species differences in diagnostic approaches, providing commentary on the benefits that might arise from simultaneous interpretation of data from human and animal health surveillance systems; and (c) reiterate the importance of using species-specific, validated diagnostic tests for surveillance and disease outbreak investigations. Emerging and endemic zoonotic diseases are likely to provide a continued threat to global health in the short- to medium-term future. A good deal of knowledge about the drivers of infectious disease emergence has been developed based on numerous examples from the recent past. Sharing of diagnostic resources across human and animal health sectors, sharing of human and animal health surveillance data, development of skills in the interpretation of that data and awareness of issues related to the validation and interpretation of diagnostic test data are necessary prerequisites for an effective endemic disease surveillance system. A good understanding of the epidemiological patterns of endemic disease will allow human and animal health professionals be able to more quickly detect the presence of emerging disease threats.

Après avoir répertorié les méthodes utilisées pour diagnostiquer les maladies zoonotiques chez l'homme comme chez les animaux, les auteurs définissent les différentes approches diagnostiques suivant les espèces considérées et commentent les avantages qui pourraient découler d'une interprétation simultanée des données par les systèmes de surveillance en santé animale et en santé publique ; il réitèrent ensuite l'importance de recourir à des tests diagnostiques validés et spécifiques de l'espèce considérée dans le cadre de la surveillance et des enquêtes suite à l'apparition d'un foyer. Les maladies zoonotiques émergentes et endémiques représentent potentiellement une menace continue pour la santé mondiale à court et à moyen terme. Les facteurs favorisant l'émergence des maladies infectieuses sont désormais beaucoup mieux connus grâce aux enseignements tirés de nombreux exemples récents. Le partage des ressources diagnostiques entre les secteurs de la santé humaine et animale, les échanges des données de la surveillance sanitaire réunies par les deux secteurs, le renforcement des compétences en matière d'interprétation des données et la sensibilisation aux problématiques de la validation et de l'interprétation des données générées par les tests de diagnostic sont des conditions préalables à la mise en place d'un système efficace de surveillance des maladies endémiques. Une bonne compréhension des profils épidémiologiques des maladies endémiques permettra aux professionnels de la santé humaine et animale de détecter plus rapidement la présence de menaces émergentes.

Los autores proceden aquí a: a) relacionar los métodos empleados para diagnosticar enfermedades zoonóticas en personas y animales; b) señalar las diferencias que existen entre los distintos planteamientos de diagnóstico según la especie de que se trate, comentando asimismo las ventajas que podrían derivarse de la interpretación simultánea de los datos de los sistemas de vigilancia sanitaria y de los de vigilancia zoosanitaria; y c) reiterar la importancia que reviste el uso de pruebas de diagnóstico no solo validadas, sino también adaptadas específicamente a cada especie, para las labores de vigilancia y estudio de brotes. Lo más probable es que a corto y medio plazo las enfermedades zoonóticas, ya sean emergentes o endémicas, sigan constituyendo una amenaza para la salud mundial. Gracias a numerosos ejemplos del pasado reciente se ha ido constituyendo un buen conocimiento de los factores que propician la aparición de enfermedades infecciosas. Para disponer de un eficaz sistema de vigilancia de enfermedades endémicas hay una serie de requisitos previos indispensables: utilización compartida de los recursos de diagnóstico entre los sectores de la salud humana y la sanidad animal; intercambio de los datos de vigilancia sanitaria y de vigilancia zoosanitaria; adquisición de competencias para interpretar esos datos; y buen conocimiento de las cuestiones ligadas a la validación de pruebas de diagnóstico y a la interpretación de los datos que arrojan. Si los profesionales de la salud humana y la sanidad animal conocen debidamente los patrones epidemiológicos de las enfermedades endémicas, estarán en condiciones de detectar con más celeridad la presencia de enfermedades emergentes que constituyan una amenaza.

A descriptive retrospective study on mortality and involuntary culling in beef and dairy cattle production systems of Western Australia (1981-2018).

Identifying and quantifying the relative frequency of involuntary losses is an essential first step in developing fit-for-purpose herd health programmes. The objective of this study was to provide an estimate of the relative frequency of reasons for mortality among south-west Western Australian beef and dairy cattle, based on necropsy findings from a university-based veterinary pathology referral centre over 38 years. A total of 904 cattle were submitted for postmortem examination throughout the study period. Gastrointestinal, cardiopulmonary and reproductive conditions were the most common causes of mortality in cattle submitted for necropsy at Murdoch University for the period 1981-2018. In dairy cattle, the common problems were gastrointestinal (bloat, abomasal displacements) 18% (59/320), cardiovascular (traumatic reticulo-pericarditis) 9% (30/320) and respiratory conditions (pneumonia) 8% (27/320). In beef cattle, the most common conditions were gastrointestinal (bloat, rumen acidosis) 11% (39/358), reproductive (metritis) 11% (38/358), cardiovascular (traumatic reticulo-pericarditis) 7% (25/358), respiratory (pneumonia) 7% (24/358), lameness (fractures) 6%, (21/358) and hepatobiliary conditions (blue-green algae poisoning, hepatotoxicity) 6% (21/358). Selection bias and missing data were potential confounders in this study. Although necropsy investigations provide useful information on animal mortalities and avenues for future herd health programmes, there is a need to standardise data capture methods and disease definition criteria, and conduct more detailed recording of data both at the farm level and at necropsy diagnostic centres.

Pet health insurance reduces the likelihood of pre-surgical euthanasia of dogs with gastric dilatation-volvulus in the emergency room of an Australian referral hospital.

AIMS: To determine the association between the presence of pet health insurance and the risk of euthanasia at the time of diagnosis for dogs with gastric dilatation-volvulus (GDV). METHODS: Insurance status at the time of GDV diagnosis was sought for a cohort of 147 non-referred, confirmed cases of GDV that presented to the emergency department of a university-based veterinary hospital in Australia between 2008 and 2017. Insurance status was obtained from the medical record (n=18) or after contacting the owners by phone using a standardised questionnaire (n=129). Animal, clinical and outcome data was retrospectively compiled in a research database. The primary outcome measure was whether or not the dog was euthanised before surgery. The Mantel-Haenszel procedure was used to quantify the association between the presence of pet health insurance and the risk of euthanasia at the time of diagnosis for dogs with GDV, adjusting for the confounding effect of age at the time of presentation using Bayesian methods. RESULTS: Of the 69 dogs for which insurance information could be obtained, 10 (14%) cases were insured at the time of the GDV event and 59 (86%) cases were not. The majority of non-insured dogs (37/59; 63 (95% CI=50-74)%) were euthanised before surgery, while none (0 (95% CI=0-28)%) of the insured dogs were euthanised at that time (p<0.001). Of the 32 insured and non-insured dogs that underwent surgery, four dogs (13 (95% CI=5-28)%) did not survive to hospital discharge. Three dogs (9%) were euthanised during or after surgery and one dog (3%) experienced cardiopulmonary arrest during treatment. The majority of dogs for which insurance status was known did not survive to hospital discharge (41/69; 59%), and 90 (95% CI=7-96)% of deaths were caused by euthanasia prior to surgery. Uninsured dogs were 5.0 (95% credible interval=1.8-26) times more likely to undergo presurgical euthanasia compared with insured dogs. CONCLUSIONS: Euthanasia prior to treatment was most common cause of death in non-referred dogs with GDV; such euthanasia was entirely absent in the cohort of dogs that were insured. CLINICAL RELEVANCE: Financial considerations significantly contribute to mortality of dogs with GDV presented to an emergency room. Financial instruments to reduce the out-of-pocket expense for pet owners confronted with unexpected veterinary expenses have potential to reduce pet mortality.

Herd manager attitudes and intentions regarding the selection of high-fertility EBV sires in Australia.

Reproductive performance in dairy cattle has declined over the last 50 years as an unintended consequence of selection for high milk yield. Since the early 2000s, dairy geneticists have released successive versions of fertility estimated breeding values (EBV) to assist in reversing this trend. At the herd level, fertility EBV can help managers accelerate improvements in reproductive performance by acting as a second selection criteria when used in tandem with a breeding index. However, use of the fertility EBV in sire selection currently varies between herd managers. The aim of this study was to better understand the reasons why herd managers choose or do not choose to select high-fertility EBV sires, using the Theory of Planned Behavior (TPB) as a social research framework. Thirty-five Victorian dairy herd managers were recruited as part of a larger study investigating the daughter fertility Australian Breeding Value and interviewed using a series of questions examining TPB constructs. The interviews were recorded and transcribed using template analysis. A wide range of herd manager types were enrolled into the study, with representation from diverse systems. Out of the 35 herd managers, 27 included fertility in their list of high-priority breeding objectives. A wide variation in results was consistent with previous studies that have demonstrated marked heterogeneity in herd manager attitudes toward bull selection. Herd manager-perceived barriers to selection of sires with high daughter fertility EBV included a lack of high daughter fertility bulls with other desirable traits, a lack of trust in the fertility EBV or in the Australian EBV system, difficulty in interpreting international proofs, information overload, semen prices, low bull reliability, and difficulty in understanding bull catalogs. Not all herd managers found the process problematic, however, particularly if a breeding consultant was employed to select all or most of the sires. Herd manager-perceived barriers for choosing to select daughter fertility as a breeding objective include a lack of awareness of the EBV, a lack of interest in genetics in general, low confidence in the impact of genetic selection for fertility, and a feeling that fertility was not important for their production system. The results of this study suggest that animal geneticists and on-farm service providers need to work together to allow the opportunities arising from appropriate use of fertility EBV to be realized more broadly across the dairy industry.

Hypoxia stimulates angiogenesis and a metabolic switch in human parathyroid adenoma cells.

Hypoxia, a primary stimulus for angiogenesis, is important for tumour proliferation and survival. The effects of hypoxia on parathyroid tumour cells, which may also be important for parathyroid autotransplantation in patients, are, however, unknown. We, therefore, assessed the effects of hypoxia on gene expression in parathyroid adenoma (PA) cells from patients with primary hyperparathyroidism. Cell suspensions from human PAs were cultured under normoxic or hypoxic conditions and then subjected to cDNA expression analysis. In total, 549 genes were significantly upregulated and 873 significantly downregulated. The most highly upregulated genes (carbonic anhydrase 9 (CA9), Solute carrier family 2A1 (SLC2A1) and hypoxia-inducible lipid droplet-associated protein (HIG2)) had known involvement in hypoxia responses. Dysregulation of oxidative phosphorylation and glycolysis pathway genes were also observed, consistent with data indicating that cells shift metabolic strategy of ATP production in hypoxic conditions and that tumour cells predominantly utilise anaerobic glycolysis for energy production. Proliferation- and angiogenesis-associated genes linked with growth factor signalling, such as mitogen-activated protein kinase kinase 1 (MAP2K1), Jun proto-oncogene (JUN) and ETS proto-oncogene 1 (ETS1), were increased, however, Ras association domain family member 1 (RASSF1), an inhibitor of proliferation was also upregulated, indicating these pathways are unlikely to be biased towards proliferation. Overall, there appeared to be a shift in growth factor signalling pathways from Jak-Stat and Ras signaling to extracellular signal-regulated kinases (ERKs) and hypoxia-inducible factor (HIF)-1α signalling. Thus, our data demonstrate that PAs, under hypoxic conditions, promote the expression of genes known to stimulate angiogenesis, as well as undergoing a metabolic switch.