Evaluation of the accuracy of the IDvet serological test for Mycoplasma bovis infection in cattle using latent class analysis of paired serum ELISA and quantitative real-time PCR on tonsillar swabs sampled at slaughter.

Mycoplasma bovis (Mbovis) was first detected in cattle in New Zealand (NZ) in July 2017. To prevent further spread, NZ launched a world-first National Eradication Programme in May 2018. Existing diagnostic tests for Mbovis have been applied in countries where Mbovis is endemic, for detecting infection following outbreaks of clinical disease. Diagnostic test evaluation (DTE) under NZ conditions was thus required to inform the Programme. We used Bayesian Latent Class Analysis on paired serum ELISA (ID Screen Mycoplasma bovis Indirect from IDvet) and tonsillar swabs (qPCR) for DTE in the absence of a gold standard. Tested samples were collected at slaughter between June 2018 and November 2019, from infected herds depopulated by the Programme. A first set of models evaluated the detection of active infection, i.e. the presence of Mbovis in the host. At a modified serology positivity threshold of SP%> = 90, estimates of animal-level ELISA sensitivity was 72.8% (95% credible interval 68.5%-77.4%), respectively 97.7% (95% credible interval 97.3%-98.1%) for specificity, while the qPCR sensitivity was 45.2% (95% credible interval 41.0%-49.8%), respectively 99.6% (95% credible interval 99.4%-99.8%) for specificity. In a second set of models, prior information about ELISA specificity was obtained from the National Beef Cattle Surveillance Programme, a population theoretically free-or very low prevalence-of Mbovis. These analyses aimed to evaluate the accuracy of the ELISA test targeting prior exposure to Mbovis, rather than active infection. The specificity of the ELISA for detecting exposure to Mbovis was 99.9% (95% credible interval 99.7%-100.0%), hence near perfect at the threshold SP%=90. This specificity estimate, considerably higher than in the first set of models, was equivalent to the manufacturer's estimate. The corresponding ELISA sensitivity estimate was 66.0% (95% credible interval 62.7%-70.7%). These results confirm that the IDvet ELISA test is an appropriate tool for determining exposure and infection status of herds, both to delimit and confirm the absence of Mbovis.

Use of scenario tree modelling to plan freedom from infection surveillance: Mycoplasma bovis in New Zealand.

Since mid-2018, the New Zealand (NZ) Ministry for Primary Industries (MPI) has been operating an eradication program for an incursion of Mycoplasma bovis. Although NZ is still delimiting the outbreak, consideration is being given to how freedom from M. bovis will be demonstrated. Rapid demonstration of freedom will minimise the length of the program, significantly reducing its financial burden. This collaborative research was undertaken to help inform planning of surveillance to demonstrate freedom after M. bovis is believed eradicated. Scenario tree modelling (STM) involves assimilating multiple surveillance system components to determine whether disease is absent. STM has infrequently been used to plan appropriate surveillance but this was the approach used here. A stochastic simulation model was implemented in R. The model represented the NZ commercial dairy and non-dairy cattle industries and the current surveillance components that are also planned to be used to gather evidence of absence of M. bovis once it is eradicated. Different surveillance intensities and risk based versus random surveillance were simulated and compared for probability of freedom, financial cost of sampling and testing and the time to demonstrate freedom. The results indicate that the current surveillance components will enable demonstration of freedom. Surveillance components included bulk tank milk testing, herd testing and testing at meat processing plants, predominantly using an imperfect ELISA. Several combinations of surveillance components appeared most efficient achieving >95 % confidence of freedom over 2-4 years, whilst sampling 4-7 % of the non-dairy herds and less than 25 % of dairy herds annually. The results indicate that surveillance intensity can be lower than is currently occurring to support the delimiting phase, thereby saving significant resources in the post eradication phase (proof of freedom phases). Further consideration is required to enable the assumption of 100 % herd specificity made in the model to be achieved. The ELISA used is very specific, but will yield some false positives that must be resolved to their true status. This may occur for example through modified diagnostic test interpretation (e.g. cut point optimisation at individual and herd level) or resolution of putative false positive herds with epidemiological investigation. In conclusion this research demonstrates the utility of STM for planning surveillance programs, and in this instance has highlighted efficient and effective surveillance components for demonstrating freedom from M. bovis in NZ. It also highlights the need to achieve 100 % specificity for M. bovis in herds tested during the proof of freedom phases.

Brucella suis Seroprevalence and Associated Risk Factors in Dogs in Eastern Australia, 2016 to 2019.

Brucella suis is a zoonotic disease of feral pigs that also affects pig hunting dogs, pig hunters, veterinarians and veterinary staff. In recent years the incidence of B. suis in the eastern Australian states of New South Wales (NSW) and Queensland (QLD) has increased. A cross-sectional study was conducted to document the seroprevalence, geographical extent and risk factors for B. suis in dogs at-risk of contracting the disease. Eligible dogs were those that were known to hunt or consume feral pig meat. Dogs were enrolled through private veterinary clinics and/or directly by District Veterinarians in six regions of NSW and QLD. Blood was collected by venepuncture and tested for B. suis antibodies using the Rose Bengal Test (RBT) followed by a Complement Fixation Test (CFT) if they returned a positive RBT. Owners were invited to complete a questionnaire on the dogs' signalment, husbandry including hunting practices and locations, and any clinical signs referable to brucellosis. Of the 317 dogs included in the prevalence survey, 21 were seropositive returning a survey-adjusted true seroprevalence of 9.3 (95% CI 0.45 to 18) B. suis positive dogs per 100 dogs at-risk. True seroprevalence ranged from 0 to 24 B. suis positive dogs per 100 across eastern Australia, with the highest prevalence in central west NSW and southern QLD. Adjusted for other factors, dogs that shared a household with other seropositive dogs and those that traveled away from their home regions to hunt were more likely to be seropositive. Clinical signs at presentation were not predictive of serostatus, with seropositive and seronegative dogs equally likely to present with signs consistent with brucellosis. The results obtained from this study show that B. suis exposure is relatively common in dogs that have contact with feral pigs, with one in 10 testing seropositive. Further studies are needed to understand the progression and risk of transmission from seropositive dogs.

Mycoplasma bovis outbreak in New Zealand cattle: An assessment of transmission trends using surveillance data.

Mycoplasma bovis most likely infected New Zealand cattle in the latter half of 2015. Infection was detected in mid-2017 after which control activities were implemented. An official eradication programme commenced in mid-2018, which is ongoing. We examined farm-level tracing and surveillance data to describe the outbreak, analyse transmission trends and make inference on progress towards eradication. Results indicate that cattle movements were the primary means of spread. Although case farms were distributed throughout both islands of New Zealand, most animal movements off infected farms did not result in newly infected farms, indicating Mycoplasma bovis is not highly transmissible between farms. To describe and analyse outbreak trends, we undertook a standard descriptive outbreak investigation, including construction of an epidemic curve and calculation of estimated dissemination ratios. We then employed three empirical models-a non-linear growth model, time series model and branching process model based on time-varying effective reproduction numbers-to further analyse transmission trends and provide short-term forecasts of farm-level incidence. Our analyses suggest that Mycoplasma bovis transmission in New Zealand has declined and progress towards eradication has been made. Few incident cases were forecast for the period between 8 September and 17 December 2019. To date, no case farms with an estimated infection date assigned to this period have been detected; however, case detection is ongoing, and these results need to be interpreted cautiously considering model validation and other important contextual information on performance of the eradication programme, such as the time between infection, detection and implementation of movement controls on case farms.

Information delivery and the veterinarian-horse owner relationship in the context of Hendra virus in Australia.

Hendra virus (HeV) is an emerging bat-borne virus endemic in Australia that can be transmitted from horses to humans and has a high fatality rate for horses and people. Controversy surrounding HeV risk mitigation measures have strained the veterinarian-horse owner relationship. This study aimed to characterise the veterinarian-horse owner relationship in general and also in the context of HeV by analysing data derived from the 'Horse Owners and Hendra Virus: A Longitudinal Study to Evaluate Risk' (HHALTER) study. Australian horse owners were recruited via emails, social media and word-of-mouth for a series of five surveys that were administered online at six-monthly intervals over a two-year period to capture baseline knowledge, attitudes and practices of horse owners regarding HeV and any changes over time. In the current study, descriptive analyses of information sources were performed to understand the use of veterinarians as a HeV information source (Surveys 1 and 5; n = 1195 and n = 617). Ordinal logistic regression analyses were conducted to determine factors associated with the frequency of horse owner contact with a veterinarian (Survey 3; n = 636). This study found a relative increase over the study period in the proportion of horse owners who had used veterinarians as HeV information source in the last 12 months (from 51.9% to 88.3%). Owning more horses, being older, having a 'duty of care' for other people working with horses and deriving the main income from horse related business were factors associated with more frequent veterinary contact. Results suggest that traditional information sources such as workshops, information packs and risk training are likely to be used by horse owners. Smart phone applications should be considered for use in the future and require further investigation for horse health communication. The findings of this study may be helpful in optimising strategies for horse health information delivery.

Efficacy of citric acid and sodium hypochlorite as disinfectants against Mycoplasma bovis.

Mycoplasma bovis, a cattle pathogen of major economic importance across the globe, causes a range of diseases, including pneumonia and mastitis. Because of the limited options for effective treatment of these diseases, prevention and control are preferred to diagnosis and treatment. In this study, the efficacies of citric acid and sodium hypochlorite as disinfectants against M. bovis were tested using a modification of a standardised method for assessing the efficacy of disinfectants against bacteria. A citric acid concentration of 0.5 % was found to be an effective disinfectant, reducing infectivity by close to 106 fold, while sodium hypochlorite at 1% was found to have similar efficacy to 0.5 % citric acid. A 0.04 % concentration of sodium hypochlorite was effective against M. bovis only in the absence of any organic material. Under these conditions, 0.25 % citric acid found to have similar efficacy. These findings indicate that 0.5 % citric acid or 1 % sodium hypochlorite are likely to be effective disinfectants for M. bovis under field conditions and 0.04 % sodium hypochlorite or 0.25 % citric acid are likely to be effective following removal of organic material.

Australian horse owners and their biosecurity practices in the context of Hendra virus.

In recent years, outbreaks of exotic as well as newly emerging infectious diseases have highlighted the importance of biosecurity for the Australian horse industry. As the first potentially fatal zoonosis transmissible from horses to humans in Australia, Hendra virus has emphasised the need to incorporate sound hygiene and general biosecurity practices into day-to-day horse management. Recommended measures are widely publicised, but implementation is at the discretion of the individual owner. This cross-sectional study aimed to determine current levels of biosecurity of horse owners and to identify factors influencing the uptake of practices utilising data from an online survey. Level of biosecurity (low, medium, high), as determined by horse owners' responses to a set of questions on the frequency of various biosecurity practices performed around healthy (9 items) and sick horses (10 items), was used as a composite outcome variable in ordinal logistic regression analyses. The majority of horse owners surveyed were female (90%), from the states of Queensland (45%) or New South Wales (37%), and were involved in either mainly competitive/equestrian sports (37%) or recreational horse activities (35%). Seventy-five percent of owners indicated that they follow at least one-third of the recommended practices regularly when handling their horses, resulting in medium to high levels of biosecurity. Main factors associated with a higher level of biosecurity were high self-rated standard of biosecurity, access to personal protective equipment, absence of flying foxes in the local area, a good sense of control over Hendra virus risk, likelihood of discussing a sick horse with a veterinarian and likelihood of suspecting Hendra virus in a sick horse. Comparison of the outcome variable with the self-rated standard of biosecurity showed that over- as well as underestimation occurred. This highlights the need for continuous communication and education to enhance awareness and understanding of what biosecurity is and how it aligns with good horsemanship. Overall, strengthened biosecurity practices will help to improve animal as well as human health and increase preparedness for future disease outbreaks.

"We've learned to live with it"-A qualitative study of Australian horse owners' attitudes, perceptions and practices in response to Hendra virus.

Hendra virus causes sporadic zoonotic disease in Australia following spill over from flying foxes to horses and from horses to people. Prevention and risk mitigation strategies such as vaccination of horses or biosecurity and property management measures are widely publicised, but hinge on initiative and action taken by horse owners as they mediate management, care and treatment of their animals. Hence, underlying beliefs, values and attitudes of horse owners influence their uptake of recommended risk mitigation measures. We used a qualitative approach to investigate attitudes, perceptions and self-reported practices of horse owners in response to Hendra virus to gain a deeper understanding of their decision-making around prevention measures. Data presented here derive from a series of in-depth interviews with 27 horse owners from Hendra virus 'hot spot' areas in New South Wales and Queensland. Interviews explored previous experience, perceptions and resulting behaviour as well as communication around Hendra virus. All interviews were recorded, transcribed verbatim and analysed in NVivo using thematic analysis. Analysis revealed four major themes: perception of Hendra virus as a risk and factors influencing this perception, Hendra virus risk mitigation strategies implemented by horse owners, perceived motivators and barriers of these strategies, and interaction of perceived risk, motivators and barriers in the decision-making process. Although Hendra virus disease was perceived as a serious threat to the health of horses and humans, individual risk perception diverged among horse owners. Perceived severity, likelihood and unpredictability as well as awareness and knowledge of Hendra virus, trust in information obtained and information pathways, demographic characteristics and personal experience were the main factors influencing Hendra virus risk perceptions. Other key determinants of horse owners' decision-making process were attitudes towards Hendra virus risk mitigation measures as well as perceived motivators and barriers thereof. Horse owners' awareness of the necessity to consider individual Hendra virus risk and adequate risk management strategies was described as a learning process, which changed over time. However, different perceptions of risk, barriers and motivators in combination with a weighing up of advantages and disadvantages resulted in different behaviours. These findings demonstrate the multifactorial determinants of cognitive mediating processes and facilitate a better understanding of horse owners' perspectives on preventive horse health measures. Furthermore, they provide valuable feedback to industry and government stakeholders on how to improve effective risk communication and encourage uptake of recommended risk mitigation measures.

An investigation of classical swine fever virus seroprevalence and risk factors in pigs in Timor-Leste.

Classical swine fever virus (CSFV) is a highly infectious pathogen of pigs and believed to be a major constraint to pig production in Timor-Leste. The Ministry of Agriculture and Fisheries conducts vaccination campaigns in an attempt to control clinical disease, however, there is no empirical data available concerning the seroprevalence and distribution of CSFV in Timor-Leste. To help address this knowledge deficit, a cross-sectional study to determine seroprevalence was conducted in the three districts that border Indonesia. Data on farmer- and pig-level factors were also collected to look at their impact on CSFV serological status. Overall, true CSFV seroprevalence was estimated at 34.4%. Seroprevalence estimates varied widely between and within districts, subdistricts, and villages. Older pigs and pigs that had been vaccinated for CSFV were more likely to test positive for CSFV antibody. Pigs owned by farmers that experienced the sudden death of pigs in the 12 months prior to the survey were more likely to test positive for CSFV antibody, while pigs that had been sick in the previous three months were less likely to test positive for CSFV antibody. The final multivariable model accounted for a large amount of variation in the data, however, much of this variation was explained by the random effects with less than one percent of the variation explained by the fixed effects. This work further supports the need for a collaborative approach to whole-island CSFV control between West Timor, Indonesia and Timor-Leste. Further work is needed to better understand the risk factors for CSFV serological status in order to allocate resources for control. As CSFV is now endemic in Timor-Leste research involving a combination of serology, antigen detection and in-depth investigation of suspect cases over a period of time may be required.

An investigation of classical swine fever virus seroprevalence and risk factors in pigs in East Nusa Tenggara, eastern Indonesia.

Classical swine fever virus (CSFV) is a highly infectious disease of pigs. It has had significant impacts on East Nusa Tenggara, eastern Indonesia since its introduction in 1997. In spite of its importance to this region, little is known about its seroprevalence and distribution, and pig-level and farmer-level factors that may have an impact on the serological status of an individual pig. To address this knowledge deficit, a cross-sectional seroprevalence survey was conducted in 2010 involving 2160 pigs and 805 farmers from four islands in the region. Farmer questionnaires and pig record forms were used to collect data about the farmers and pigs surveyed. Blood was collected from each pig to determine its CSFV serological status. Apparent and true prevalence were calculated for each island, district, subdistrict, and village surveyed. CSFV serological status was used as an outcome variable in mixed effects logistic regression analyses. Overall true CSFV seroprevalence was estimated at 17.5% (lower CI 16.0%; upper CI 19.5%). Seroprevalence estimates varied widely across the islands, districts, subdistricts, and villages. Manggarai Barat, a district on the western end of Flores Island, contained pigs that were positive for antibody to CSFV. This result was unexpected, as no clinical cases had been reported in this area. Older pigs and pigs that had been vaccinated for CSFV were more likely to test positive for antibody to CSFV. The final multivariable model accounted for a large amount of variation in the data, however much of this variation was explained by the random effects with less than 2% of the variation explained by pig age and pig CSFV vaccination status. In this study we documented the seroprevalence of CSFV across four islands in East Nusa Tenggara, eastern Indonesia. We also identified risk factors for the presence of antibody to CSFV. Further investigation is needed to understand why clinical CSFV has not been reported on the western end of Flores Island, and to identify additional risk factors that explain CSFV serological status to inform disease control strategies.

The use of a modified Delphi approach to engage stakeholders in zoonotic disease research priority setting.

BACKGROUND: After the 2011 cluster of Hendra virus cases in horses in Australia, public health targeted education initiatives at people in the equine industry to reduce human exposure to potentially infected horses. 'Horse owners and Hendra Virus: A Longitudinal cohort study To Evaluate Risk' aims to enhance public health measures through improved understanding of Hendra virus risk perception and risk mitigation strategies among horse owners and horse care providers. This paper describes the stakeholder consultation that was undertaken to ensure the cohort study outcomes were relevant to diverse groups who play a role in Hendra virus policy development and implementation. METHODS: A two-round modified Delphi study with online questionnaires was conducted. In round one, stakeholders identified priority research areas. In round two, stakeholders rated and ranked topics that emerged from thematic analysis of the round one responses. Round two data were analysed using logistic regression. RESULTS: Of the 255 stakeholders contacted, 101 responded to round one. Over 450 topics were proposed. These were organized into 18 themes. Approximately two thirds of the round one respondents participated in round two. 'Hendra virus-related risk awareness and perception', 'personal health and safety', 'emergency preparedness', 'risk prevention, mitigation, and biosecurity', and 'Hendra virus vaccination in horses--attitudes/uptake' were the top five areas identified according to probability of being ranked extremely important. CONCLUSIONS: In this study, a modified Delphi approach was effective in guiding research into Hendra virus, a zoonotic disease of animal and human health significance. The findings support the notion that stakeholders should be engaged in zoonotic disease research priority setting. Such consultation will help to ensure that research initiatives are relevant and useful to stakeholders in the position to make use of new findings.

A focused ethnographic study of Alberta cattle veterinarians' decision making about diagnostic laboratory submissions and perceptions of surveillance programs.

The animal and public health communities need to address the challenge posed by zoonotic emerging infectious diseases. To minimize the impacts of future events, animal disease surveillance will need to enable prompt event detection and response. Diagnostic laboratory-based surveillance systems targeting domestic animals depend in large part on private veterinarians to submit samples from cases to a laboratory. In contexts where pre-diagnostic laboratory surveillance systems have been implemented, this group of veterinarians is often asked to input data. This scenario holds true in Alberta where private cattle veterinarians have been asked to participate in the Alberta Veterinary Surveillance Network-Veterinary Practice Surveillance, a platform to which pre-diagnostic disease and non-disease case data are submitted. Consequently, understanding the factors that influence these veterinarians to submit cases to a laboratory and the complex of factors that affect their participation in surveillance programs is foundational to interpreting disease patterns reported by laboratories and engaging veterinarians in surveillance. A focused ethnographic study was conducted with ten cattle veterinarians in Alberta. Individual in-depth interviews with participants were recorded and transcribed to enable thematic analysis. Laboratory submissions were biased toward outbreaks of unknown cause, cases with unusual mortality rates, and issues with potential herd-level implications. Decreasing cattle value and government support for laboratory testing have contributed to fewer submissions over time. Participants were willing participants in surveillance, though government support and collaboration were necessary. Changes in the beef industry and veterinary profession, as well as cattle producers themselves, present both challenges and opportunities in surveillance.

A focused ethnographic study of Sri Lankan government field veterinarians' decision making about diagnostic laboratory submissions and perceptions of surveillance.

The global public health community is facing the challenge of emerging infectious diseases. Historically, the majority of these diseases have arisen from animal populations at lower latitudes where many nations experience marked resource constraints. In order to minimize the impact of future events, surveillance of animal populations will need to enable prompt event detection and response. Many surveillance systems targeting animals rely on veterinarians to submit cases to a diagnostic laboratory or input clinical case data. Therefore understanding veterinarians' decision-making process that guides laboratory case submission and their perceptions of infectious disease surveillance is foundational to interpreting disease patterns reported by laboratories and engaging veterinarians in surveillance initiatives. A focused ethnographic study was conducted with twelve field veterinary surgeons that participated in a mobile phone-based surveillance pilot project in Sri Lanka. Each participant agreed to an individual in-depth interview that was recorded and later transcribed to enable thematic analysis of the interview content. Results found that field veterinarians in Sri Lanka infrequently submit cases to laboratories--so infrequently that common case selection principles could not be described. Field veterinarians in Sri Lanka have a diagnostic process that operates independently of laboratories. Participants indicated a willingness to take part in surveillance initiatives, though they highlighted a need for incentives that satisfy a range of motivations that vary among field veterinarians. This study has implications for the future of animal health surveillance, including interpretation of disease patterns reported, system design and implementation, and engagement of data providers.

A hidden Markov model for analysis of frontline veterinary data for emerging zoonotic disease surveillance.

Surveillance systems tracking health patterns in animals have potential for early warning of infectious disease in humans, yet there are many challenges that remain before this can be realized. Specifically, there remains the challenge of detecting early warning signals for diseases that are not known or are not part of routine surveillance for named diseases. This paper reports on the development of a hidden Markov model for analysis of frontline veterinary sentinel surveillance data from Sri Lanka. Field veterinarians collected data on syndromes and diagnoses using mobile phones. A model for submission patterns accounts for both sentinel-related and disease-related variability. Models for commonly reported cattle diagnoses were estimated separately. Region-specific weekly average prevalence was estimated for each diagnoses and partitioned into normal and abnormal periods. Visualization of state probabilities was used to indicate areas and times of unusual disease prevalence. The analysis suggests that hidden Markov modelling is a useful approach for surveillance datasets from novel populations and/or having little historical baselines.

Mobile phone-based infectious disease surveillance system, Sri Lanka.

Because many infectious diseases are emerging in animals in low-income and middle-income countries, surveillance of animal health in these areas may be needed for forecasting disease risks to humans. We present an overview of a mobile phone-based frontline surveillance system developed and implemented in Sri Lanka. Field veterinarians reported animal health information by using mobile phones. Submissions increased steadily over 9 months, with ≈4,000 interactions between field veterinarians and reports on the animal population received by the system. Development of human resources and increased communication between local stakeholders (groups and persons whose actions are affected by emerging infectious diseases and animal health) were instrumental for successful implementation. The primary lesson learned was that mobile phone-based surveillance of animal populations is acceptable and feasible in lower-resource settings. However, any system implementation plan must consider the time needed to garner support for novel surveillance methods among users and stakeholders.