Evaluation of Ki-67, goblet cell and MUC2 mucin RNA expression in dogs with lymphoplasmacytic and granulomatous colitis.

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% did not correlate with colonic inflammation overall. Median MUC2% normalised to WHS in the NC group (10.02%; range, 3.05-39.09%) was two- and three-fold higher than in the GC and LPC groups respectively. With increased colonic inflammation, despite minimal changes in GBC% overall, MUC2 expression markedly declined in the LPC group (-27.4%; 95%-CI, -49.8, 5.9%) and mildly declined in the GC and NC groups. Granulomatous colitis and LPC likely involve different pathways regulating MUC2 expression. Decreased MUC2 gene expression is observed in dogs with chronic colitis compared to dogs without colonic signs. Changes in MUC2 expression appear influenced by GBC activity rather than quantity in GC and LPC.

Testing the intrinsic mechanisms driving the dynamics of Ross River Virus across Australia.

The mechanisms driving dynamics of many epidemiologically important mosquito-borne pathogens are complex, involving combinations of vector and host factors (e.g., species composition and life-history traits), and factors associated with transmission and reporting. Understanding which intrinsic mechanisms contribute most to observed disease dynamics is important, yet often poorly understood. Ross River virus (RRV) is Australia's most important mosquito-borne disease, with variable transmission dynamics across geographic regions. We used deterministic ordinary differential equation models to test mechanisms driving RRV dynamics across major epidemic centers in Brisbane, Darwin, Mandurah, Mildura, Gippsland, Renmark, Murray Bridge, and Coorong. We considered models with up to two vector species (Aedes vigilax, Culex annulirostris, Aedes camptorhynchus, Culex globocoxitus), two reservoir hosts (macropods, possums), seasonal transmission effects, and transmission parameters. We fit models against long-term RRV surveillance data (1991-2017) and used Akaike Information Criterion to select important mechanisms. The combination of two vector species, two reservoir hosts, and seasonal transmission effects explained RRV dynamics best across sites. Estimated vector-human transmission rate (average ß = 8.04x10-4per vector per day) was similar despite different dynamics. Models estimate 43% underreporting of RRV infections. Findings enhance understanding of RRV transmission mechanisms, provide disease parameter estimates which can be used to guide future research into public health improvements and offer a basis to evaluate mitigation practices.

Exposure to Persistent Organic Pollutants in Australian Waterbirds.

There is growing worldwide recognition of the threat posed by persistent organic pollutants (POPs) to wildlife populations. We aimed to measure exposure levels to POPs in a Southern Hemisphere aquatic waterbird species, the nomadic gray teal (Anas gracilis), which is found across Australia. We collected wings from 39 ducks harvested by recreational hunters at two sites (one coastal, one inland) in Victoria, southeastern Australia, in 2021. We examined three groups of POPs: nine congeners of polychlorinated biphenyls (PCBs), 13 organochlorine pesticides (OCPs), and 12 polycyclic aromatic hydrocarbons (PAHs). The PCBs, OCPs, and PAHs were detected at quantifiable levels in 13%, 72%, and 100% of birds, respectively. Of the congeners we tested for in PCBs, OCPs, and PAHs, 33%, 38%, and 100% were detected at quantifiable levels, respectively. The highest levels of exposure to POPs that we found were to the PAH benzo[b]fluoranthene, occurring at a concentration range of 1.78 to 161.05 ng/g wet weight. There were some trends detected relating to differences between geographical sites, with higher levels of several PAHs at the coastal versus inland site. There were several strong, positive associations among PAHs found. We discuss potential sources for the POPs detected, including industrial and agricultural sources, and the likely role of large-scale forest fires in PAH levels. Our results confirm that while Australian waterbirds are exposed to a variety of POPs, exposure levels are currently relatively low. Additional future investigations are required to further characterize POPs within Australian waterbird species. Environ Toxicol Chem 2024;43:736-747. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Source attribution of campylobacteriosis in Australia, 2017-2019.

Campylobacter jejuni and Campylobacter coli infections are the leading cause of foodborne gastroenteritis in high-income countries. Campylobacter colonizes a variety of warm-blooded hosts that are reservoirs for human campylobacteriosis. The proportions of Australian cases attributable to different animal reservoirs are unknown but can be estimated by comparing the frequency of different sequence types in cases and reservoirs. Campylobacter isolates were obtained from notified human cases and raw meat and offal from the major livestock in Australia between 2017 and 2019. Isolates were typed using multi-locus sequence genotyping. We used Bayesian source attribution models including the asymmetric island model, the modified Hald model, and their generalizations. Some models included an "unsampled" source to estimate the proportion of cases attributable to wild, feral, or domestic animal reservoirs not sampled in our study. Model fits were compared using the Watanabe-Akaike information criterion. We included 612 food and 710 human case isolates. The best fitting models attributed >80% of Campylobacter cases to chickens, with a greater proportion of C. coli (>84%) than C. jejuni (>77%). The best fitting model that included an unsampled source attributed 14% (95% credible interval [CrI]: 0.3%-32%) to the unsampled source and only 2% to ruminants (95% CrI: 0.3%-12%) and 2% to pigs (95% CrI: 0.2%-11%) The best fitting model that did not include an unsampled source attributed 12% to ruminants (95% CrI: 1.3%-33%) and 6% to pigs (95% CrI: 1.1%-19%). Chickens were the leading source of human Campylobacter infections in Australia in 2017-2019 and should remain the focus of interventions to reduce burden.

Field epidemiology capacity of the national veterinary services of Lao PDR: An online survey.

Introduction: Capacity in veterinary epidemiology is critical to detect, respond and control infectious diseases. Laos veterinary service is limited by having only a small number of veterinarians who graduated from overseas. Animal science graduates support the majority of the Laos veterinary services. The veterinary program was established in 2009 at the National University of Laos. We aimed to understand the national veterinary epidemiology capacity and identify gaps and training needs. Method: A cross-sectional online study was conducted in 2021 targeting central (DLF), provincial (PAFO) and district (DAFO) government animal health officers and veterinary/animal science academics (n = 332). The questionnaire addressed skills, experiences and perceived training needs in outbreak investigation, disease surveillance, data management and analysis, epidemiological surveys, One Health, leadership and communication and biosecurity. A descriptive analysis was performed and associations between demographic factors and epidemiological skills were examined. Results and discussion: In total, 205 respondents completed the questionnaire (61.8% response rate). Respondents reported low or no skills and experience in data management and analysis, epidemiological surveys and One Health. In contrast, higher but limited skills and experiences were reported in outbreak investigation, disease surveillance and biosecurity. Previous epidemiology training was primarily associated with stronger experiences in veterinary epidemiology competencies, followed by respondents that had completed a veterinary degree, highlighting the value of the currently available epidemiology training and veterinary-trained personnel in Lao PDR. This study could help inform the Laos government in its policy planning for field veterinary epidemiology capacity and future training.

A repeatable CT protocol for quantifying caudal vena cava growth in medium and large breed dogs.

Developmental malformations can cause stunted or abnormal growth and clinical disease in dogs. In humans, measurements of the inferior vena cava are used as methods for detecting abnormal growth trajectories. The objectives of this retrospective, multicenter, analytical, cross-sectional study were to develop a repeatable protocol to measure the caudal vena cava (CVC) and generate growth curves in medium and large-breed dogs during development. Contrast-enhanced CT DICOM images from 438 normal dogs, aged from 1 to 18 months, from five specific breeds were included. A "best guess" measurement protocol was created. Dogs were stratified into medium or large breed groups based on growth rate trajectories. Linear regression models and logarithmic trend lines were used to evaluate the CVC growth over time. The CVC measurements were analyzed from four anatomical regions: thorax, diaphragm, intra-hepatic, and renal. The thoracic segment produced the most repeatable measurements with the highest explanatory power. The CVC thoracic circumference ranged from 2.5 to 4.9 cm from 1 to 18 months of age. Medium and large breeds had similar CVC growth trajectories, with comparable estimated marginal means, however medium dogs reached 80% of predicted final CVC size approximately 4 weeks earlier than large breed dogs. This new protocol provides a standardized technique for evaluation of the CVC circumference over time using contrast-enhanced CT and is most repeatable when taken at the thoracic level. This approach could be adapted for other vessels to predict their growth trajectories, generating healthy reference population data for comparison against patients with vascular anomalies.

Estimation of a Within-Herd Transmission Rate for African Swine Fever in Vietnam.

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.

Lead contamination in Australian game meat.

Lead-based ammunition (gunshot and bullets) frequently leaves small lead fragments embedded in the meat of wild-shot game animals. Australia produces several commercial game meat products from wild animals harvested with lead-based ammunition and has a growing population of recreational hunters. However, no studies have previously investigated the frequency of lead fragments or lead concentrations in Australian game meat. We examined 133 Australian minced game meat items of four types for evidence of lead contamination. Samples were meat from kangaroos (Macropus and Osphranter spp.; n=36) and Bennett's wallabies (Notamacropus rufogriseus; n=28) sold for human consumption, and deer ('venison'; multiple spp.; n=32) and stubble quail (Coturnix pectoralis; n=37) harvested for private consumption by recreational hunters. All packages were studied by digital radiography to detect the presence of radio-dense fragments, assumed to be lead fragments from ammunition. Visible fragments were absent in commercially available kangaroo products, but were present in 4%, 28% and 35% of wallaby, venison and quail, respectively. Mean meat lead concentrations (mg/kg wet weight) were 0.01 ± 0.01 for kangaroo, 0.02 ± 0.01 for wallaby, 0.12 ± 0.07 for venison, and 1.76 ± 3.76 for quail. The Australian food standards threshold for livestock meat (0.1 mg/kg w.w.) was not exceeded by any kangaroo or wallaby products but was exceeded by 53% and 86% of venison and quail, respectively. Radiography only detected 35% of samples that were above the food safety threshold. While average lead concentrations in commercially available macropod (kangaroo and wallaby) meat were low, those in recreationally harvested game meat may pose health risks for hunters and associated consumers.

Performance Evaluation and Validation of Air Samplers To Detect Aerosolized Coxiella burnetii.

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, BioSampler, and Coriolis Micro) for their efficacy in detecting C. burnetii. Our results show that all three devices could detect aerosolized bacteria effectively; however, at high concentrations the AirPort performed better than the other two devices, showing higher percent recovery. At intermediate and low concentrations AirPort detected at a level higher than or similar to that of other samplers. Quantification of samples was hindered by the limit of quantitation of the qPCR assay. Compared with the other two devices, the AirPort was easier to handle and clean in the field. Testing air around likely sources (e.g., farms, abattoirs, and livestock saleyards) using validated sampling devices will help better estimate the risk of Q fever to nearby communities.

Validation of an Indirect Immunofluorescence Assay and Commercial Q Fever Enzyme-Linked Immunosorbent Assay for Use in Macropods.

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.

Risk factors for campylobacteriosis in Australia: outcomes of a 2018-2019 case-control study.

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 consumption of chicken meat. However, contact with young pet dogs may also be an important source of infection. Proton-pump inhibitors are likely to increase vulnerability to infection.

Bayesian latent class analysis to estimate the optimal cut-off for the MilA ELISA for the detection of Mycoplasma bovis antibodies in sera, accounting for repeated measures.

The MilA ELISA has been identified as a highly effective diagnostic tool for the detection of Mycoplasma bovis specific antibodies and has been validated for serological use in previous studies. This study aimed to estimate the optimal cut-off and corresponding estimates of diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the MilA ELISA for testing bovine serum. Serum samples from 298 feedlot cattle from 14 feedlots across four Australian states were tested on entry into the feedlot and approximately 42 days later. The paired serum samples were tested with the MilA ELISA, BIO K302 (Bio-X Diagnostics, Belgium) and BIO K260 (Bio-X Diagnostics, Belgium). A cut-off of 135 AU was estimated to be optimal using Bayesian latent class analysis with three tests in multiple populations, accounting for conditional dependence between tests. At this cut-off, the DSe and DSp of the MilA ELISA were estimated to be 92.1 % (95 % highest probability density [HPD] interval: 87.4, 95.8) and 95.5 % (95 % HPD: 92.4, 97.8), respectively. The DSes of the BIO K260 and BIO K302 ELISAs were estimated to be 60.5 % (95 % HPD: 54.0, 66.9) and 44.6 % (95 % HPD: 38.7, 50.7), respectively. DSps were 95.6 % (95 % HPD: 92.9, 97.7) and 97.8 % (95 % HPD: 95.9, 99.0), respectively. Mycoplasma bovis seroprevalence was remarkably high at follow-up after 42 days on the feedlots. Overall, this study estimated a cut-off, DSe and DSp for the MilA ELISA with less dependence on prior information than previous analyses and demonstrated that the MilA ELISA has higher DSe than the BIO K260 and BIO K302 assays.

Evaluation of the MilA ELISA for the diagnosis of herd infection with Mycoplasma bovis using bulk tank milk and estimation of the prevalence of M. bovis in Australia.

Infection with Mycoplasma bovis has been identified as a growing threat in dairy industries worldwide and there is an urgent need for an inexpensive and accurate herd-level screening tool to identify herds that have been exposed to M. bovis. This study aimed to evaluate the use of the MilA ELISA for testing bulk tank milk (BTM) samples for antibodies against M. bovis and estimate a suitable cut-off and diagnostic sensitivity (DSe) and specificity (DSp) for this assay. An optimal cut-off was then applied for investigating the geographical and seasonal distribution of infection with M. bovis in Australia. A total of 5554 BTM samples from 2683 dairy herds were collected during March, August and December 2017. BTM samples were tested in the MilA ELISA and a cut-off of 29 antibody units (AU) was estimated to be optimal using Bayesian latent class analysis which makes no assumption about the true disease status of herds under investigation. At this cut-off, the DSe and DSp were estimated to be 96.6% (95% highest probability density [HPD] interval: 87.0, 99.8) and 94.2% (95% HPD: 89.9, 97.4), respectively. The diagnostic specifications were found to vary markedly with stage of the production cycle, suggesting that targeted sampling was needed to maximize accuracy. We also found distinct differences in the apparent prevalence of M. bovis in different dairying regions, as well as seasonal variation. The highest apparent prevalence of M. bovis was observed in samples collected in March and an overall drop in the proportion of positive herds was seen from March to December. Overall, this study provides insights into the dynamics of BTM antibodies against M. bovis in Australian dairy herds and how the MilA ELISA can be applied for bulk tank milk testing.

Identifying scenarios and risk factors for Q fever outbreaks using qualitative analysis of expert opinion.

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.

The Potential Role of Wild Suids in African Swine Fever Spread in Asia and the Pacific Region.

African swine fever (ASF) in Asia and the Pacific is currently dominated by ASF virus transmission within and between domestic pig populations. The contribution made by wild suids is currently not well understood; their distribution, density and susceptibility to the virus has raised concerns that their role in the epidemiology of ASF in the region might be underestimated. Whilst in the Republic of Korea wild suids are considered important in the spread and maintenance of ASF virus, there is an apparent underreporting to official sources of the disease in wild suids from other countires and regions. A review of the current literature, an analysis of the official reporting resources and a survey of the World Organisation of Animal Health Member delegates in Asia and the Pacific were used to assess the potential role of wild suids in ASF outbreaks, and also to gain insight into what ASF management or control strategies are currently implemented for wild suids. Applying appropriate population control and management strategies can be increased in some areas, especially to assist in the conservation of endangered endemic wild suids in this region.

A randomised controlled trial of the immunogenicity and safety of a formaldehyde-inactivated Coxiella burnetii vaccine in 8-week-old goats.

Coxiella burnetii causes Q fever in individuals exposed to infected ruminants. Vaccination in 3-4-month-old goats, has been reported to result in significantly greater reduction in C. burnetii shedding compared to goats vaccinated one month before breeding, the most commonly used strategy of controlling Q fever on infected intensively-managed herds. It is possible that an even greater reduction in the number of animals shedding C. burnetii could be achieved if vaccination were administered shortly after protection from colostrum antibodies wanes and animals become susceptible to infection with C. burnetii. This study aimed to evaluate the immunogenicity and safety of a formaldehyde-inactivated phase 1 C. burnetii vaccine in 8-week-old goats. Two injections, four weeks apart, elicited specific IgM and IgG responses in all vaccinated goats (n = 6), while no antibodies were detected in two control groups (n = 12). Swelling at the site of inoculation was observed in all the vaccinated and in 10/11 of the placebo-treated goats but receded after 3 weeks. Weight change and rectal temperatures were also comparable between vaccinated and control goats. The data indicated that this vaccine could be suitable for immunising 8-week-old goats, although further trials to determine level of protection against challenge are required.

Optimising predictive modelling of Ross River virus using meteorological variables.

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 predicting mosquito-borne disease notifications and outbreaks used for disease surveillance.

Using farmer observations for animal health syndromic surveillance: Participation and performance of an online enhanced passive surveillance system.

The challenge of animal health surveillance is to provide the information necessary to appropriately inform disease prevention and control activities within the constraints of available resources. Syndromic surveillance of farmers' disease observations can improve animal health data capture from extensive livestock farming systems, especially where data are not otherwise being systematically collected or when data on confirmed aetiological diagnoses are unavailable at the disease level. As it is rarely feasible to recruit a truly random sample of farmers to provide observational reports, directing farmer sampling to align with the surveillance objectives is a reasonable and practical approach. As long as potential bias is recognised and managed, farmers who will report reliably can be desirable participants in a surveillance system. Thus, one early objective of a surveillance program should be to identify characteristics associated with reporting behaviour. Knowledge of the demographic and managerial characteristics of good reporters can inform efforts to recruit additional farms into the system or aid understanding of potential bias of system reports. We describe the operation of a farmer syndromic surveillance system in Victoria, Australia, over its first two years from 2014 to 2016. Survival analysis and classification and regression tree analysis were used to identify farm level factors associated with 'reliable' participation (low non-response rates in longitudinal reporting). Response rate and timeliness were not associated with whether farmers had disease to report, or with different months of the year. Farmers keeping only sheep were the most reliable and timely respondents. Farmers < 43 years of age had lower response rates than older farmers. Farmers with veterinary qualifications and those working full-time on-farm provided less timely reports than other educational backgrounds and farmers who worked part-time on-farm. These analyses provide a starting point to guide recruitment of participants for surveillance of farmers' observations using syndromic surveillance, and provide examples of strengths and weaknesses of syndromic surveillance systems for extensively-managed livestock. Once farm characteristics associated with reliable participation are known, they can be incorporated into surveillance system design in accordance with the objectives of the system.

Coxiella burnetii in the environment: A systematic review and critical appraisal of sampling methods.

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 involve environmental sampling for C. burnetii.

Serological evidence for the presence of wobbly possum disease virus in Australia.

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.