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Intestinal mucus barrier disruption may occur with chronic inflammatory enteropathies. The lack of studies evaluating mucus health in dogs with chronic colitis arises from inherent challenges with assessment of the intestinal mucus layer. It is therefore unknown if reduced goblet cell (GBC) numbers and/or mucin 2 (MUC2) expression, which are responsible for mucus production and secretion, correlate with inflammation severity in dogs with granulomatous colitis (GC) or lymphocytic-plasmacytic colitis (LPC). It is undetermined if Ki-67 immunoreactivity, which has been evaluated in dogs with small intestinal inflammation, similarly correlates to histologic severity in GC and LPC. Study objectives included comparing Ki-67 immunoreactivity, GBC population and MUC2 expression in dogs with GC, LPC and non-inflamed colon; and exploring the use of ribonucleic acid (RNAscope®) in-situ hybridization (ISH) to evaluate MUC2 expression in canine colon. Formalin-fixed endoscopic colonic biopsies were obtained from 48 dogs over an eight-year period. A blinded pathologist reviewed all biopsies. Dogs were classified into the GC (n=19), LPC (n=19) or no colitis (NC) (n=10) group based on final histopathological diagnosis. Ki-67 immunohistochemistry, Alcian-Blue/PAS staining to highlight GBCs, and RNAscope® ISH using customized canine MUC2-targeted probes were performed. At least five microscopic fields per dog were selected to measure Ki-67 labelling index (KI67%), GBC staining percentage (GBC%) and MUC2 expression (MUC2%) using image analysis software. Spearman's correlation coefficients were used to determine associations between World Small Animal Veterinary Association histologic score (WHS) and measured variables. Linear regression models were used to compare relationships between WHS with KI67%, GBC%, and MUC2%; and between GBC% and MUC2%. Median WHS was highest in dogs with GC. Median KI67% normalised to WHS was highest in the NC group (6.69%; range, 1.70-23.60%). Median GBC% di

We describe results from a panel study in which pigs from a 17-sow African swine fever (ASF) positive herd in Thái Bình province, Vietnam, were followed over time to record the date of onset of ASF signs and the date of death from ASF. Our objectives were to (1) fit a susceptible-exposed-infectious-removed disease model to the data with transmission coefficients estimated using approximate Bayesian computation; (2) provide commentary on how a model of this type might be used to provide decision support for disease control authorities. For the outbreak in this herd, the median of the average latent period was 10 days (95% HPD (highest posterior density interval): 2 to 19 days), and the median of the average duration of infectiousness was 3 days (95% HPD: 2 to 4 days). The estimated median for the transmission coefficient was 3.3 (95% HPD: 0.4 to 8.9) infectious contacts per ASF-infectious pig per day. The estimated median for the basic reproductive number, R0, was 10 (95% HPD: 1.1 to 30). Our estimates of the basic reproductive number R0 were greater than estimates of R0 for ASF reported previously. The results presented in this study may be used to estimate the number of pigs expected to be showing clinical signs at a given number of days following an estimated incursion date. This will allow sample size calculations, with or without adjustment to account for less than perfect sensitivity of clinical examination, to be used to determine the appropriate number of pigs to examine to detect at least one with the disease. A second use of the results of this study would be to inform the equation-based within-herd spread components of stochastic agent-based and hybrid simulation models of ASF.

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.

BACKGROUND: We aimed to identify risk factors for sporadic campylobacteriosis in Australia, and to compare these for Campylobacter jejuni and Campylobacter coli infections. METHODS: In a multi-jurisdictional case-control study, we recruited culture-confirmed cases of campylobacteriosis reported to state and territory health departments from February 2018 through October 2019. We recruited controls from notified influenza cases in the previous 12 months that were frequency matched to cases by age group, sex, and location. Campylobacter isolates were confirmed to species level by public health laboratories using molecular methods. We conducted backward stepwise multivariable logistic regression to identify significant risk factors. RESULTS: We recruited 571 cases of campylobacteriosis (422 C. jejuni and 84 C. coli) and 586 controls. Important risk factors for campylobacteriosis included eating undercooked chicken (adjusted odds ratio [aOR] 70, 95% CI 13-1296) or cooked chicken (aOR 1.7, 95% CI 1.1-2.8), owning a pet dog aged < 6 months (aOR 6.4, 95% CI 3.4-12), and the regular use of proton-pump inhibitors in the 4 weeks prior to illness (aOR 2.8, 95% CI 1.9-4.3). Risk factors remained similar when analysed specifically for C. jejuni infection. Unique risks for C. coli infection included eating chicken pâté (aOR 6.1, 95% CI 1.5-25) and delicatessen meats (aOR 1.8, 95% CI 1.0-3.3). Eating any chicken carried a high population attributable fraction for campylobacteriosis of 42% (95% CI 13-68), while the attributable fraction for proton-pump inhibitors was 13% (95% CI 8.3-18) and owning a pet dog aged < 6 months was 9.6% (95% CI 6.5-13). The population attributable fractions for these variables were similar when analysed by campylobacter species. Eating delicatessen meats was attributed to 31% (95% CI 0.0-54) of cases for C. coli and eating chicken pâté was attributed to 6.0% (95% CI 0.0-11). CONCLUSIONS: The main risk factor for campylobacteriosis in Australia is consum

Q fever is an important zoonotic disease perceived to be an occupational hazard for those working with livestock. Outbreaks involving large numbers of people are uncommon, but the increasing case incidence coupled with changing environmental and industry conditions that promote transmission of Q fever has raised concerns that large and serious outbreaks could become more frequent. The aim of this study was to use expert opinion to better understand how large Q fever outbreaks might occur in an Australian context and to document factors believed to be drivers of disease transmission. Focus groups were conducted with human and animal health professionals across several Australian states. All discussions were recorded, transcribed verbatim and imported into NVIVO for thematic analysis. Four anthropogenic risk factors (disease awareness, industry practices, land use, human behaviour) and three ecological risk factors (physical environment, agent dissemination, animal hosts) emerged from the data. Analysis of expert opinions pointed to the existence of numerous scenarios in which Q fever outbreaks could occur, many of which depict acquisition in the wider community outside of traditional at-risk occupations. This perception of the expansion of Q fever from occupational-acquisition to community-acquisition is driven by greater overarching economic, political and socio-cultural influences that govern the way in which people live and work. Findings from this study highlight that outbreaks are complex phenomena that involve the convergence of diverse elements, not just that of the pathogen and host, but also the physical, political and socioeconomic environments in which they interact. A review of the approaches to prevent and manage Q fever outbreaks will require a multisectorial approach and strengthening of community education, communication and engagement so that all stakeholders become an integrated part of outbreak mitigation and response.

Kangaroos are considered to be an important reservoir of Q fever in Australia, although there is limited knowledge on the true prevalence and distribution of coxiellosis in Australian macropod populations. Serological tests serve as useful surveillance tools, but formal test validation is needed to be able to estimate true seroprevalence rates, and few tests have been validated to screen wildlife species for Q fever. In this study, we modified and optimized a phase-specific indirect immunofluorescence assay (IFA) for the detection of IgG antibodies against Coxiella burnetii in macropod sera. The assay was validated against the commercially available ID Screen Q fever indirect multispecies enzyme-linked immunosorbent assay (ELISA) kit (IDVet, Grabels, France) to estimate the diagnostic sensitivity and specificity of each assay, using Bayesian latent class analysis. A direct comparison of the two tests was performed by testing 303 serum samples from 10 macropod populations from the east coast of Australia and New Zealand. The analysis indicated that the IFA had relatively high diagnostic sensitivity (97.6% [95% credible interval [CrI], 88.0 to 99.9]) and diagnostic specificity (98.5% [95% CrI, 94.4 to 99.9]). In comparison, the ELISA had relatively poor diagnostic sensitivity (42.1% [95% CrI, 33.7 to 50.8]) and similar diagnostic specificity (99.2% [95% CrI, 96.4 to 100]) using the cutoff values recommended by the manufacturer. The estimated true seroprevalence of C. burnetii exposure in the macropod populations included in this study ranged from 0% in New Zealand and Victoria, Australia, up to 94.2% in one population from New South Wales, Australia.

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.

Coxiella burnetii, the etiological agent of Q fever, is an intracellular zoonotic pathogen transmitted via the respiratory route. Once released from infected animals, C. burnetii can travel long distances through air before infecting another host. As such, the ability to detect the presence of C. burnetii in air is important. In this study, three air samplers, AirPort MD8, BioSampler, and the Coriolis Micro, were assessed against a set of predetermined criteria in the presence of three different aerosolized C. burnetii concentrations. Two liquid collection media, phosphate-buffered saline (PBS) and alkaline polyethylene glycol (Alk PEG), were tested with devices requiring a collection liquid. Samples were tested by quantitative polymerase chain reaction assay (qPCR) targeting the single-copy com1 gene or multicopy insertion element IS1111. All air samplers performed well at detecting airborne C. burnetii across the range of concentrations tested. At high nebulized concentrations, AirPort MD8 showed higher, but variable, recovery probabilities. While the BioSampler and Coriolis Micro recovered C. burnetii at lower concentrations, the replicates were far more repeatable. At low and intermediate nebulized concentrations, results were comparable in the trials between air samplers, although the AirPort MD8 had consistently higher recovery probabilities. In this first study validating air samplers for their ability to detect aerosolized C. burnetii, we found that while all samplers performed well, not all samplers were equal. It is important that these results are further validated under field conditions. These findings will further inform efforts to detect airborne C. burnetii around known point sources of infection. IMPORTANCE Coxiella burnetii causes Q fever in humans and coxiellosis in animals. It is important to know if C. burnetii is present in the air around putative sources as it is transmitted via inhalation. This study assessed air samplers (AirPort MD8, BioSamp

BACKGROUND: Statistical models are regularly used in the forecasting and surveillance of infectious diseases to guide public health. Variable selection assists in determining factors associated with disease transmission, however, often overlooked in this process is the evaluation and suitability of the statistical model used in forecasting disease transmission and outbreaks. Here we aim to evaluate several modelling methods to optimise predictive modelling of Ross River virus (RRV) disease notifications and outbreaks in epidemiological important regions of Victoria and Western Australia. METHODOLOGY/PRINCIPAL FINDINGS: We developed several statistical methods using meteorological and RRV surveillance data from July 2000 until June 2018 in Victoria and from July 1991 until June 2018 in Western Australia. Models were developed for 11 Local Government Areas (LGAs) in Victoria and seven LGAs in Western Australia. We found generalised additive models and generalised boosted regression models, and generalised additive models and negative binomial models to be the best fit models when predicting RRV outbreaks and notifications, respectively. No association was found with a model's ability to predict RRV notifications in LGAs with greater RRV activity, or for outbreak predictions to have a higher accuracy in LGAs with greater RRV notifications. Moreover, we assessed the use of factor analysis to generate independent variables used in predictive modelling. In the majority of LGAs, this method did not result in better model predictive performance. CONCLUSIONS/SIGNIFICANCE: We demonstrate that models which are developed and used for predicting disease notifications may not be suitable for predicting disease outbreaks, or vice versa. Furthermore, poor predictive performance in modelling disease transmissions may be the result of inappropriate model selection methods. Our findings provide approaches and methods to facilitate the selection of the best fit statistical model for pr

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.

Q fever is a zoonotic disease caused by the intracellular bacterium, Coxiella burnetii. Its primary mode of transmission is by inhalation of aerosols originating from infected animals and contaminated environments. The organism has a very low infective dose, can persist in the environment for long periods of time and large outbreaks fuelled by windborne spread have been previously reported. Detection of C. burnetii in the environment is therefore important during human and animal outbreak investigations and for the control and prevention of Q fever. This study aimed to systematically review and critically appraise the published literature on sampling methods used to detect C. burnetii from different environmental samples. A search of four electronic databases with subsequent hand searching identified 47 eligible articles published since 1935. These articles described sampling of dust, air, soil and liquids in attempts to detect C. burnetii during 19 Q fever outbreaks and in 28 endemic settings. Environmental positivity was most commonly associated with ruminant livestock populations. Evidence describing spatio-temporal characteristics and associated geographical dispersion gradients was limited. The most commonly tested sample type was dust which also yielded the highest bacterial loads of >108 bacteria/cloth. The MD8 (Sartorius) air sampler was used widely for air sampling. Soil was the only sample type for which a validated laboratory protocol was established specifically for C. burnetii. Each environmental sample type has its advantages and limitations which are discussed in detail and a simplified framework to guide decisions around environmental sampling for C. burnetii is provided. In any type of environmental sampling, it is recommended to use standardized and validated methods and to match the most ideal sampling strategy and timing with the research context. These conditions are essential to be considered when designing future Q fever management plans that

The aim of this study was to describe the spatiotemporal distribution of H5 HPAI outbreak reports for the period 2014-2017 and to identify factors associated with H5 HPAI outbreak reports. Throughout the study period, a total of 139 outbreaks of H5 HPAI in poultry were reported, due to either H5N1 (96 outbreaks) or H5N6 (43 outbreaks) subtype viruses. H5N1 HPAI outbreaks occurred in all areas of Vietnam while H5N6 HPAI outbreaks were only reported in the northern and central provinces. We counted the number of H5N1 and H5N6 outbreak report-positive districts per province over the four-year study period and calculated the provincial-level standardized morbidity ratio for H5N1 and H5N6 outbreak reports as the observed number of positive districts divided by the expected number. A mixed-effects, zero-inflated Poisson regression model was developed to identify risk factors for outbreak reports of each H5N1 and H5N6 subtype virus. Spatially correlated and uncorrelated random effects terms were included in this model to identify areas of the country where outbreak reports occurred after known risk factors had been accounted-for. The presence of an outbreak report in a province in the previous 6-12 months increased the provincial level H5N1 outbreak report risk by a factor of 2.42 (95% Bayesian credible interval [CrI] 1.27-4.60) while 1000 bird increases in the density of chickens decreased provincial level H5N6 outbreak report risk by a factor of 0.65 (95% CrI 0.38 to 0.97). We document distinctly different patterns in the spatial and temporal distribution of H5N1 and H5N6 outbreak reports. Most of the variation in H5N1 report risk was accounted-for by the fixed effects included in the zero-inflated Poisson model. In contrast, the amount of unaccounted-for risk in the H5N6 model was substantially greater than the H5N1 model. For H5N6 we recommend that targeted investigations should be carried out in provinces with relatively large spatially correlated random effect terms

Ross River virus (RRV) is Australia's most epidemiologically important mosquito-borne disease. During RRV epidemics in the State of Victoria (such as 2010/11 and 2016/17) notifications can account for up to 30% of national RRV notifications. However, little is known about factors which can forecast RRV transmission in Victoria. We aimed to understand factors associated with RRV transmission in epidemiologically important regions of Victoria and establish an early warning forecast system. We developed negative binomial regression models to forecast human RRV notifications across 11 Local Government Areas (LGAs) using climatic, environmental, and oceanographic variables. Data were collected from July 2008 to June 2018. Data from July 2008 to June 2012 were used as a training data set, while July 2012 to June 2018 were used as a testing data set. Evapotranspiration and precipitation were found to be common factors for forecasting RRV notifications across sites. Several site-specific factors were also important in forecasting RRV notifications which varied between LGA. From the 11 LGAs examined, nine experienced an outbreak in 2011/12 of which the models for these sites were a good fit. All 11 LGAs experienced an outbreak in 2016/17, however only six LGAs could predict the outbreak using the same model. We document similarities and differences in factors useful for forecasting RRV notifications across Victoria and demonstrate that readily available and inexpensive climate and environmental data can be used to predict epidemic periods in some areas. Furthermore, we highlight in certain regions the complexity of RRV transmission where additional epidemiological information is needed to accurately predict RRV activity. Our findings have been applied to produce a Ross River virus Outbreak Surveillance System (ROSS) to aid in public health decision making in Victoria.

Transmission network modelling to infer ‘who infected whom’ in infectious disease outbreaks is a highly active area of research. Outbreaks of foot-and-mouth disease have been a key focus of transmission network models that integrate genomic and epidemiological data. The aim of this study was to extend Lau’s systematic Bayesian inference framework to incorporate additional parameters representing predominant species and numbers of animals held on a farm. Lau’s Bayesian Markov chain Monte Carlo algorithm was reformulated, verified and pseudo-validated on 100 simulated outbreaks populated with demographic data Japan and Australia. The modified model was then implemented on genomic and epidemiological data from the 2010 outbreak of foot-and-mouth disease in Japan, and outputs compared to those from the SCOTTI model implemented in BEAST2. The modified model achieved improvements in overall accuracy when tested on the simulated outbreaks. When implemented on the actual outbreak data from Japan, infected farms that held predominantly pigs were estimated to have five times the transmissibility of infected cattle farms and be 49% less susceptible. The farm-level incubation period was 1 day shorter than the latent period, the timing of the seeding of the outbreak in Japan was inferred, as were key linkages between clusters and features of farms involved in widespread dissemination of this outbreak. To improve accessibility the modified model has been implemented as the R package ‘BORIS’ for use in future outbreaks.

Q fever, caused by the zoonotic bacterium Coxiella burnetii, is a globally distributed emerging infectious disease. Livestock are the most important zoonotic transmission sources, yet infection in people without livestock exposure is common. Identifying potential exposure pathways is necessary to design effective interventions and aid outbreak prevention. We used natural language processing and graphical network methods to provide insights into how Q fever notifications are associated with variation in patient occupations or lifestyles. Using an 18-year time-series of Q fever notifications in Queensland, Australia, we used topic models to test whether compositions of patient answers to follow-up exposure questionnaires varied between demographic groups or across geographical areas. To determine heterogeneity in possible zoonotic exposures, we explored patterns of livestock and game animal co-exposures using Markov Random Fields models. Finally, to identify possible correlates of Q fever case severity, we modelled patient probabilities of being hospitalized as a function of particular exposures. Different demographic groups consistently reported distinct sets of exposure terms and were concentrated in different areas of the state, suggesting the presence of multiple transmission pathways. Macropod exposure was commonly reported among Q fever cases, even when exposure to cattle, sheep or goats was absent. Males, older patients and those that reported macropod exposure were more likely to be hospitalized due to Q fever infection. Our study indicates that follow-up surveillance combined with text modelling is useful for unravelling exposure pathways in the battle to reduce Q fever incidence and associated morbidity.

Wobbly possum disease virus (WPDV) is an arterivirus that was originally identified in common brushtail possums (Trichosurus vulpecula) in New Zealand, where it causes severe neurological disease. In this study, serum samples (n = 188) from Australian common brushtail, mountain brushtail (Trichosurus cunninghami) and common ringtail (Pseudocheirus peregrinus) possums were tested for antibodies to WPDV using ELISA. Antibodies to WPDV were detected in possums from all three species that were sampled in the states of Victoria and South Australia. Overall, 16% (30/188; 95% CI 11.0-22.0) of possums were seropositive for WPDV and 11.7% (22/188; 95% CI 7.5-17.2) were equivocal. The frequency of WPDV antibody detection was the highest in possums from the two brushtail species. This is the first reported serological evidence of infection with WPDV, or an antigenically similar virus, in Australian possums, and the first study to find antibodies in species other than common brushtail possums. Attempts to detect viral RNA in spleens by PCR were unsuccessful. Further research is needed to characterise the virus in Australian possums and to determine its impact on the ecology of Australian marsupials.

Toxoplasma gondii is considered a disease risk for many native Australian species. Feral cats are the key definitive host of T. gondii in Australia and therefore, investigating the epidemiology of T. gondii in cat populations is essential to understanding the risk posed to wildlife. Test sensitivity and specificity are poorly defined for diagnostic tests targeting T. gondii in cats and there is a need for validated techniques. This study focused on the feral cat population on Phillip Island, Victoria, Australia. We compared a novel real-time PCR (qPCR) protocol to the modified agglutination test (MAT) and used a Bayesian latent class modelling approach to assess the diagnostic parameters of each assay and estimate the true prevalence of T. gondii in feral cats. In addition, we performed multivariable logistic regression to determine risk factors associated with T. gondii infection in cats. Overall T. gondii prevalence by qPCR and MAT was 79.5% (95% confidence interval 72.6-85.0) and 91.8% (84.6-95.8), respectively. Bayesian modelling estimated the sensitivity and specificity of the MAT as 96.2% (95% credible interval 91.8-98.8) and 82.1% (64.9-93.6), and qPCR as 90.1% (83.6-95.5) and 96.0% (82.1-99.8), respectively. True prevalence of T. gondii infection in feral cats on Phillip Island was estimated as 90.3% (83.2-95.1). Multivariable logistic regression analysis indicated that T. gondii infection was positively associated with weight and this effect was modified by season. Cats trapped in winter had a high probability of infection, regardless of weight. The present study suggests qPCR applied to tissue is a highly sensitive, specific and logistically feasible tool for T. gondii testing in feral cat populations. Additionally, T. gondii infection is highly prevalent in feral cats on Phillip Island, which may have significant impacts on endemic and introduced marsupial populations.

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hooved animals. Global outbreaks have highlighted the significant economic, trade, psychosocial and animal welfare impacts that can arise from the detection of disease in previously 'FMD-free' countries. Rapid and early diagnosis provides significant advantages in disease control and minimization of deleterious consequences. We describe the process of further development and validation of a reverse-transcription loop-mediated isothermal amplification foot-and-mouth disease virus (RT-LAMP-FMDV) test, using a published LAMP primer set, for use in the field. An internal positive control (IPC) was designed and introduced for use with the assay to mitigate any intrinsic interference from the unextracted field samples and avoid false negatives. Further modifications were included to improve the speed and operability of the test, for use by non-laboratory trained staff operating under field conditions, with shelf-stable reaction kits which require a minimum of liquid handling skills. Comparison of the assay performance with an established laboratory-based real-time reverse transcriptase PCR (rRT-PCR) test targeting the 3D region of FMD virus (Tetracore Inc) was investigated. LAMP has the potential to complement current laboratory diagnostics, such as rRT-PCR, as a preliminary tool in the investigation of FMD. We describe a strategic approach to validation of the test for use in the field using extracted RNA samples of various serotypes from Thailand and then finally unextracted field samples collected from FMD-suspected animals (primarily oral lesion swabs) from Bhutan and Australia. The statistical approach to validation was performed by Frequentist and Bayesian latent class methods, which both confirmed this new RT-LAMP-FMDV test as fit-for-purpose as a herd diagnostic tool with diagnostic specificity >99% and sensitivity 79% (95% Bayesian credible interval: 65, 90%) on unextracted field samples

Between 1991 and 2014 the per capita notification rate of salmonellosis in Australia increased from 31.9 to 69.7 cases per 100,000 people. Salmonella Typhimurium accounted for nearly half the human cases until the end of 2014. In this study, we used cluster analysis tools to compare S. Typhimurium isolates from a chicken-meat study with those reported to the National Enteric Pathogen Surveillance System (NEPSS) from the coincident human and non-human populations. There was limited phage type diversity within all populations and a lack of specificity of MLVA profiling within phage types. The chicken-meat study isolates were not significantly clustered with the human cases and at least 7 non-human sources, based on typing profiles (PT/MLVA combination), could be implicated as a source of human cases during the same period. In the absence of a strong surveillance system representative of all putative sources, MLVA and phage typing alone or in combination are insufficient to identify the source of human cases.

A number of transmission network models are available that combine genomic and epidemiological data to reconstruct networks of who infected whom during infectious disease outbreaks. For such models to reliably inform decision-making they must be transparently validated, robust, and capable of producing accurate predictions within the short data collection and inference timeframes typical of outbreak responses. A lack of transparent multi-model comparisons reduces confidence in the accuracy of transmission network model outputs, negatively impacting on their more widespread use as decision-support tools. We undertook a formal comparison of the performance of nine published transmission network models based on a set of foot-and-mouth disease outbreaks simulated in a previously free country, with corresponding simulated phylogenies and genomic samples from animals on infected premises. Of the transmission network models tested, Lau's systematic Bayesian integration framework was found to be the most accurate for inferring the transmission network and timing of exposures, correctly identifying the source of 73% of the infected premises (with 91% accuracy for sources with model support >0.80). The Structured COalescent Transmission Tree Inference provided the most accurate inference of molecular clock rates. This validation study points to which models might be reliably used to reconstruct similar future outbreaks and how to interpret the outputs to inform control. Further research could involve extending the best-performing models to explicitly represent within-host diversity so they can handle next-generation sequencing data, incorporating additional animal and farm-level covariates and combining predictions using Ensemble methods and other approaches.

This study aimed to elucidate virus, host and environmental dynamics of Vietnamese H5 highly pathogenic avian influenza viruses (HPAIVs) during 2014-2017. Epidemiologically, H5 HPAIVs were frequently detected in apparently healthy domestic and Muscovy ducks and therefore these are preferred species for H5 HPAIV detection in active surveillance. Virologically, clade 2.3.2.1c and 2.3.4.4 H5 HPAIVs were predominant and exhibited distinct phylogeographic evolution. Clade 2.3.2.1c viruses clustered phylogenetically in North, Central and South regions, whilst clade 2.3.4.4 viruses only detected in North and Central regions formed small groups. These viruses underwent diverse reassortment with existence of at least 12 genotypes and retained typical avian-specific motifs. These H5 HPAIVs exhibited large antigenic distance from progenitor viruses and commercial vaccines currently used in poultry. Bayesian phylodynamic analysis inferred that clade 2.3.2.1c viruses detected during 2014-2017 were likely descended from homologous clade viruses imported to Vietnam previously and/or preexisting Chinese viruses during 2012-2013. Vietnamese clade 2.3.4.4 viruses closely shared genetic traits with contemporary foreign spillovers, suggesting that there existed multiple transboundary virus dispersals to Vietnam. This study provides insights into the evolution of Vietnamese H5 HPAIVs and highlights the necessity of strengthening control measures such as, preventive surveillance and poultry vaccination.

Cat-borne parasites and their associated diseases have substantial impacts on human, livestock, and wildlife health worldwide. Despite this, large and detailed datasets that allow researchers to study broad-scale trends in the ecology of cat-borne diseases are either difficult to obtain or non-existent. One condition that is easily detected at slaughter is macroscopic sarcocystosis, a cat-borne parasitosis of sheep (Ovis aries). We conducted a cross-sectional study to describe the geographic distribution of sarcocystosis in sheep throughout South Australia and investigate ecosystem characteristics associated with the presence of disease. Data were obtained from two slaughterhouses which processed 3,865,608 sheep from 4,204 farms across 385,468 km2 of South Australia's land mass for the period 2007–2017. A Poisson point process model was developed to quantify environmental characteristics associated with higher densities of sarcocystosis-positive farms. Sarcocystosis was highly clustered on a large island off of the Australian coast and the density of sarcocystosis-positive farms increased in areas of low soil pH (intensity ratio: 0.86, 95% CI: 0.78, 0.95) and high clay content. We hypothesize that region was confounded by, and predominately acted as a proxy for, cat density. Our results have broader implications regarding the health, welfare, economic, and conservation impacts of other cat-borne parasitosis, such as toxoplasmosis.

Research aimed at understanding transmission networks, representing a network of “who infected whom” for an infectious disease outbreak, have been actively conducted in recent years. Transmission network models incorporating epidemiological and genetic data are valuable for elucidating disease transmission pathways. In this study, we reconstructed the transmission network of the foot-and-mouth disease (FMD) epidemic in Japan in 2010, and explored farm-level risk factors associated with increased risk of secondary transmission. A published, systematic Bayesian transmission network model was applied to epidemiological data of 292 infected farms and whole genome sequence data of 104 of the infected farms. This model can make inferences for known infected farms even lacking genetic data. After estimating the consensus network, the accuracy of the network was examined by comparison with epidemiological data. Then, risk factors inferred to have been sources of secondary transmission were explored using zero-inflated Poisson regression model. As far as we are aware, this study represents the largest FMD outbreak transmission network to be published by such means combining epidemiological and genetic data. The consensus network reasonably generated the epidemiological links, which were estimated from the actual epidemiological investigation. Among 292 farms, 101 farms (35%) were inferred to have been the sources of secondary transmission, and amongst these farms, the median number of secondary cases was 2 (min:1–max:18) farms. The farm-type (small and large -sized pig farms), the number of days from onset to notification, and the number of susceptible farms within a 1-km radius were significantly associated with secondary transmission. Transmission network modelling enabled inference of the connections between infected farms during the FMD epidemic and identified important factors for controlling the risk of secondary transmission. This study demonstrated that the predomina

To better understand factors influencing infectious agent dispersal within a livestock population information is needed on the nature and frequency of contacts between farm enterprises. This study uses social network analysis to describe the contact network within a vertically integrated broiler poultry enterprise to identify the potential horizontal and vertical transmission pathways for Salmonella spp. Nodes (farms, sheds, production facilities) were identified and the daily movement of commodities (eggs, birds, feed, litter) and people between nodes were extracted from routinely kept farm records. Three time periods were examined in detail, 1- and 8- and 17-weeks of the production cycle and contact networks were described for all movements, and by commodity and production type. All nodes were linked by at least one movement during the study period but network density was low indicating that all potential pathways between nodes did not exist. Salmonella spp. transmission via vertical or horizontal pathways can only occur along directed pathways when those pathways are present. Only two locations (breeder or feed nodes) were identified where the transmission of a single Salmonella spp. clone could theoretically percolate through the network to the broiler or processing nodes. Only the feed transmission pathway directly connected all parts of the network.

Background This was a panel study of the prevalence of C. burnetii infection in does in an endemic dairy goat enterprise in Victoria, Australia. Our first objective was to determine the prevalence of does shedding C. burnetii at the time of parturition and to quantify the concentration of genome equivalents (GE) present in each C. burnetii positive sample. Our second objective was to determine the proportion of positive does that were persistent shedders. Our final objective was to quantify the association between C. burnetii qPCR status at the time of kidding and daily milk volumes produced during the subsequent lactation. Results Vaginal swabs (n= 490) were collected from does at the time of kidding and analysed using a quantitative polymerase chain reaction (qPCR) assay. Shedding of C. burnetii was detected in 15% (95% CI: 12% to 18%) of the sampled does. Does were classified as qPCR-negative, qPCR-positive low and qPCR-positive high based on the estimated concentration of GE from the qPCR. Persistent shedding at relatively low concentrations was detected in 20% (95% CI: 10% to35%) of shedding does sampled again at their subsequent parturition. After controlling for possible confounders and adjusting for variation in daily milk yields at the individual doe level, daily milk yields for qPCR-positive high does were reduced by 17% (95% CI: 3% to 32%) compared to qPCR-negative does (p= 0.02). Conclusions Shedding concentrations of C. burnetii were highly skewed, with a relatively small group of does shedding relatively high quantities of C. burnetii. Further, high shedding does had reduced milk yields compared to qPCR-negative does. Early detection and culling of high shedding does would result in increased farm profitability and reduce the risk of Q fever transmission.

OBJECTIVE: To describe the epidemiological and clinical features of acute Q fever in Victoria from 1994 to 2013. DESIGN: Retrospective case series and spatiotemporal analyses of human notification data. METHODS: Records for all confirmed cases of Q fever in Victoria notified between 1994 and 2013 were reviewed. Clinical and epidemiological features of the cases were described and spatiotemporal analysis undertaken for all cases potentially acquired within Victoria. RESULTS: A total of 659 confirmed acute Q fever cases were notified over the study period. Cases decreased at a rate of 4.2% per annum (95% confidence interval (CI): 0.9, 7.4%). Notification rates decreased among abattoir workers and related occupations by 10.9% per annum (95% CI: 6.5, 15.0%), whereas those among dairy farmers rose by 14.9% per annum (95% CI: 4.7, 26.0%). The mean age of cases increased over the study period while the ratio of male to female cases decreased. Spatiotemporal analysis suggested endemic transmission, with 55% of cases associated with abattoirs and related businesses and a further 30% considered to have acquired the infection locally. In addition to abattoir-associated clusters, important foci for local acquisition included South and East Gippsland, Wodonga and an outbreak centred on a dairy goat farm west of Melbourne. CONCLUSIONS: There has been a reduction in cases of acute Q fever in Victoria over the past 20 years and a changing epidemiology with respect to age, sex and acquisition source. Epidemiological and spatiotemporal analyses suggested a low level of endemic transmission within the state, with multiple foci of increased zoonotic transmission.

Mast cell tumours (MCTs) are common tumours of the canine skin, and are estimated to represent up to 20% of all skin tumours in dogs. Tumour grade has a major impact on the incidence of local recurrence and metastatic potential. In addition to helping the clinician with surgical planning, knowledge of the tumour grade also assists in proper prognostication and client education. For pre-treatment biopsies to be useful, there must exist a high level of correlation between the histopathological grade obtained from the pre-treatment biopsy and the actual histopathological grade from the excisional biopsy. The aim of this study was to determine concordance of tumour grade between various biopsy techniques (wedge, punch, needle core) and the "gold standard" excisional biopsy method. We found an overall concordance rate of 96% based on the Patnaik grading system, and an overall concordance rate of 92% based on the Kiupel grading system. The accuracy of the various biopsy techniques (wedge, punch and needle core) when compared with excisional biopsy was 92%, 100% and 100%, respectively, based on the Patnaik grading system, and 90%, 95% and 100%, respectively, based on the Kiupel grading system. Of the cases with discordant results, the pre-treatment biopsies tended to underestimate the grade of the tumour. Based on these results, we conclude that pre-treatment biopsies are sufficiently accurate for differentiating low-grade from high-grade MCTs, regardless of biopsy technique or tumour location.

BACKGROUND: Acute polyradiculoneuritis (APN) is an immune-mediated peripheral nerve disorder in dogs that shares many similarities with Guillain-Barré syndrome (GBS) in humans, in which the bacterial pathogen Campylobacter spp. now is considered to be a major triggering agent. Little information is available concerning the relationship between APN and Campylobacter spp. in dogs. HYPOTHESIS/OBJECTIVES: To estimate the association between Campylobacter spp. infection and APN. Associations with additional potential risk factors also were investigated, particularly consumption of raw chicken. ANIMALS: Twenty-seven client-owned dogs suffering from suspected APN and 47 healthy dogs, client-owned or owned by staff members. METHODS: Case-control study with incidence density-based sampling. Fecal samples were collected from each enrolled animal to perform direct culture, DNA extraction, and polymerase chain reaction (PCR) for detection of Campylobacter spp. In some cases, species identification was performed by sequence analysis of the amplicon. Data were obtained from the medical records and owner questionnaires in both groups. RESULTS: In cases in which the fecal sample was collected within 7 days from onset of clinical signs, APN cases were 9.4 times more likely to be positive for Campylobacter spp compared to control dogs (P < 0.001). In addition, a significant association was detected between dogs affected by APN and the consumption of raw chicken (96% of APN cases; 26% of control dogs). The most common Campylobacter spp. identified was Campylobacter upsaliensis. CONCLUSIONS AND CLINICAL IMPORTANCE: Raw chicken consumption is a risk factor in dogs for the development of APN, which potentially is mediated by infection with Campylobacter spp.

The transmission of Salmonella enterica within a vertically integrated poultry operation was investigated longitudinally over an 18-month period (2013-2014). Thirty six percent of all samples collected (1503 of 4219) were positive for salmonellae with seven Salmonella enterica subsp. enterica serovars, and one Salmonella enterica subsp. salamae serovar detected. Both Salmonella enterica subsp. enterica serovars Infantis and Typhimurium were detected in all locations sampled. Salmonella Typhimurium was the most frequently detected serovar (63% of serotyped samples) with 8 phage types (PT) and 41 multiple-locus variable-number tandem-repeats analysis (MLVA) profiles identified. The most frequently identified phage types were PT135a and DT135. A total of 62 PT/MLVA combinations were observed. MLVA profiles 03-14-10-09-525 and 03-15-11-11-525 were the most frequently identified and 83% of the isolates shared at least one MLVA profile with an isolate from another phage type. The use of phage typing and MLVA profiling, on their own or in combination, were insufficient to understand the complexity of the epidemiological relationships between locations within this production system. Despite the high level of apparent diversity, cluster analysis was unable to differentiate the transmission pathways of all S. Typhimurium variants detected within the integrated enterprise. Using additional epidemiological information, the parent breeder rearing site was identified as the most likely point of introduction of two S. Typhimurium isolates into the production system with subsequent dissemination to the broiler flocks via the hatchery. This complexity is unable to be resolved in the absence of intensive sampling programs at all generations of the production system.

BACKGROUND: Educational initiatives targeting at-risk populations have long been recognized as a mainstay of ongoing rabies control efforts. Cluster-based studies are often utilized to assess levels of knowledge, attitudes and practices of a population in response to education campaigns. The design of cluster-based studies requires estimates of intra-cluster correlation coefficients obtained from previous studies. This study estimates the school-level intra-cluster correlation coefficient (ICC) for rabies knowledge change following an educational intervention program. METHODS: A cross-sectional survey was conducted with 226 students from 7 schools in Sikkim, India, using cluster sampling. In order to assess knowledge uptake, rabies education sessions with pre- and post-session questionnaires were administered. Paired differences of proportions were estimated for questions answered correctly. A mixed effects logistic regression model was developed to estimate school-level and student-level ICCs and to test for associations between gender, age, school location and educational level. RESULTS: The school- and student-level ICCs for rabies knowledge and awareness were 0.04 (95% CI: 0.01, 0.19) and 0.05 (95% CI: 0.2, 0.09), respectively. These ICCs suggest design effect multipliers of 5.45 schools and 1.05 students per school, will be required when estimating sample sizes and designing future cluster randomized trials. There was a good baseline level of rabies knowledge (mean pre-session score 71%), however, key knowledge gaps were identified in understanding appropriate behavior around scared dogs, potential sources of rabies and how to correctly order post rabies exposure precaution steps. After adjusting for the effect of gender, age, school location and education level, school and individual post-session test scores improved by 19%, with similar performance amongst boys and girls attending schools in urban and rural regions. The proportion of participants that were ab

Ross River virus (RRV) is a mosquito-borne virus endemic to Australia. The disease, marked by arthritis, myalgia and rash, has a complex epidemiology involving several mosquito species and wildlife reservoirs. Outbreak years coincide with climatic conditions conducive to mosquito population growth. We developed regression models for human RRV notifications in the Mildura Local Government Area, Victoria, Australia with the objective of increasing understanding of the relationships in this complex system, providing trigger points for intervention and developing a forecast model. Surveillance, climatic, environmental and entomological data for the period July 2000-June 2011 were used for model training then forecasts were validated for July 2011-June 2015. Rainfall and vapour pressure were the key factors for forecasting RRV notifications. Validation of models showed they predicted RRV counts with an accuracy of 81%. Two major RRV mosquito vectors (Culex annulirostris and Aedes camptorhynchus) were important in the final estimation model at proximal lags. The findings of this analysis advance understanding of the drivers of RRV in temperate climatic zones and the models will inform public health agencies of periods of increased risk.

In Vietnam, live bird markets are found in most populated centres, providing the means by which fresh poultry can be purchased by consumers for immediate consumption. Live bird markets are aggregation points for large numbers of poultry, and therefore, it is common for a range of avian influenza viruses to be mixed within live bird markets as a result of different poultry types and species being brought together from different geographical locations. We conducted a cross-sectional study in seven live bird markets in four districts of Thua Thien Hue Province in August and December, 2014. The aims of this study were to (i) document the prevalence of avian influenza in live bird markets (as measured by virus isolation); and (ii) quantify individual bird-, seller- and market-level characteristics that rendered poultry more likely to be positive for avian influenza virus at the time of sale. A questionnaire soliciting details of knowledge, attitude and avian influenza practices was administered to poultry sellers in study markets. At the same time, swabs and faecal samples were collected from individual poultry and submitted for isolation of avian influenza virus. The final data set comprised samples from 1,629 birds from 83 sellers in the seven live bird markets. A total of 113 birds were positive for virus isolation; a prevalence of 6.9 (95% CI 5.8-8.3) avian influenza virus-positive birds per 100 birds submitted for sale. After adjusting for clustering at the market and individual seller levels, none of the explanatory variables solicited in the questionnaire were significantly associated with avian influenza virus isolation positivity. The proportions of variance at the individual market, seller and individual bird levels were 6%, 48% and 46%, respectively. We conclude that the emphasis of avian influenza control efforts in Vietnam should be at the individual seller level as opposed to the market level.

Although many studies have reported the indirect immunofluorescence assay (IFA) to be more sensitive in detection of antibodies to Coxiella burnetii than the complement fixation test (CFT), the diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the assay have not been previously established for use in ruminants. This study aimed to validate the IFA by describing the optimization, selection of cutoff titers, repeatability, and reliability as well as the DSe and DSp of the assay. Bayesian latent class analysis was used to estimate diagnostic specifications in comparison with the CFT and the enzyme-linked immunosorbent assay (ELISA). The optimal cutoff dilution for screening for IgG and IgM antibodies in goat serum using the IFA was estimated to be 1:160. The IFA had good repeatability (>96.9% for IgG, >78.0% for IgM), and there was almost perfect agreement (Cohen's kappa > 0.80 for IgG) between the readings reported by two technicians for samples tested for IgG antibodies. The IFA had a higher DSe (94.8%; 95% confidence interval [CI], 80.3, 99.6) for the detection of IgG antibodies against C. burnetii than the ELISA (70.1%; 95% CI, 52.7, 91.0) and the CFT (29.8%; 95% CI, 17.0, 44.8). All three tests were highly specific for goat IgG antibodies. The IFA also had a higher DSe (88.8%; 95% CI, 58.2, 99.5) for detection of IgM antibodies than the ELISA (71.7%; 95% CI, 46.3, 92.8). These results underscore the better suitability of the IFA than of the CFT and ELISA for detection of IgG and IgM antibodies in goat serum and possibly in serum from other ruminants.

Governments are routinely involved in the biosecurity of agricultural and food imports and exports. This involves controlling the complex ongoing threat of the broad range of zoonoses: endemic, exotic and newly emerging. Policy-related decision-making in these areas requires accurate information and predictions concerning the effects and potential impacts of zoonotic diseases. The aim of this article was to provide information concerning the development and use of utility-based tools, specifically disability-adjusted life years (DALYs), for measuring the burden on human disease (morbidity and mortality) as a consequence of zoonotic infections. Issues and challenges to their use are also considered. Non-monetary utility approaches that are reviewed in this paper form one of a number of tools that can be used to estimate the monetary and non-monetary 'cost' of morbidity- and mortality-related consequences. Other tools derive from cost-of-illness, willingness-to-pay and multicriteria approaches. Utility-based approaches are specifically designed to capture the pain, suffering and loss of functioning associated with diseases, zoonotic and otherwise. These effects are typically complicated to define, measure and subsequently 'cost'. Utility-based measures will not be able to capture all of the effects, especially those that extend beyond the health sector. These will more normally be captured in financial terms. Along with other uncommon diseases, the quality of the relevant epidemiological data may not be adequate to support the estimation of losses in utility as a result of zoonoses. Other issues in their use have been identified. New empirical studies have shown some success in addressing these issues. Other issues await further study. It is concluded that, bearing in mind all caveats, utility-based methods are important tools in assessing the magnitude of the impacts of zoonoses in human disease. They make an important contribution to decision-making and priority set