Veterinary Biosciences - Research Publications

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    Performance Evaluation and Validation of Air Samplers To Detect Aerosolized Coxiella burnetii
    Abeykoon, AMH ; Poon, M ; Firestone, SM ; Stevenson, MA ; Wiethoelter, AK ; Vincent, GA ; Uzal, F (AMER SOC MICROBIOLOGY, 2022-10-26)
    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.
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    Estimation of a Within-Herd Transmission Rate for African Swine Fever in Vietnam
    Le, VP ; Lan, NT ; Canevari, JT ; Villanueva-Cabezas, JP ; Padungtod, P ; Trinh, TBN ; Nguyen, VT ; Pfeiffer, CN ; Oberin, MV ; Firestone, SM ; Stevenson, MA (MDPI, 2023-02)
    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.
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    A cross-sectional survey of risk factors for the presence of Coxiella burnetii in Australian commercial dairy goat farms
    Hou, KW ; Wiethoelter, AK ; Stevenson, MA ; Soares Magalhaes, RJ ; Lignereux, L ; Caraguel, C ; Stenos, J ; Vincent, G ; Aleri, JW ; Firestone, SM (WILEY, 2022-07)
    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.
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    Validation of an Indirect Immunofluorescence Assay and Commercial Q Fever Enzyme-Linked Immunosorbent Assay for Use in Macropods
    Tolpinrud, A ; Stenos, J ; Chaber, A-L ; Devlin, JM ; Herbert, C ; Pas, A ; Dunowska, M ; Stevenson, MA ; Firestone, SM ; Barrs, VR (AMER SOC MICROBIOLOGY, 2022-07-20)
    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.
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    Identifying scenarios and risk factors for Q fever outbreaks using qualitative analysis of expert opinion
    Tan, TS-E ; Hernandez-Jover, M ; Hayes, LM ; Wiethoelter, AK ; Firestone, SM ; Stevenson, MA ; Heller, J (WILEY, 2022-06)
    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.
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    Using farmer observations for animal health syndromic surveillance: Participation and performance of an online enhanced passive surveillance system
    Pfeiffer, C ; Stevenson, M ; Firestone, S ; Larsen, J ; Campbell, A (ELSEVIER, 2021-03)
    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.
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    Spatiotemporal and risk analysis of H5 highly pathogenic avian influenza in Vietnam, 2014-2017
    Lam, TN ; Stevenson, MA ; Firestone, SM ; Sims, LD ; Duc, HC ; Long, VN ; Tien, NN ; Kien, TL ; Isoda, N ; Matsuno, K ; Okamatsu, M ; Kida, H ; Sakoda, Y (ELSEVIER, 2020-05)
    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 to identify likely determinants of disease. Similarly, investigations should be carried out in provinces with relatively low spatially correlated random effect terms to identify protective factors for disease and/or reasons for failure to report.
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    Coxiella burnetii in the environment: A systematic review and critical appraisal of sampling methods
    Abeykoon, AMH ; Clark, NJ ; Soares Magalhaes, RJ ; Vincent, GA ; Stevenson, MA ; Firestone, SM ; Wiethoelter, AK (WILEY, 2021-05)
    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.
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    Unravelling animal exposure profiles of human Q fever cases in Queensland, Australia, using natural language processing
    Clark, NJ ; Tozer, S ; Wood, C ; Firestone, SM ; Stevenson, M ; Caraguel, C ; Chaber, A-L ; Heller, J ; Magalhaes, RJS (WILEY-HINDAWI, 2020-09)
    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.
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    Factors associated with racing performance and career duration for Victorian-born Thoroughbreds
    Flash, ML ; Crabb, HK ; Hitchens, PL ; Firestone, SM ; Stevenson, MA ; Gilkerson, JR (WILEY, 2022-01)
    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.