A model for the assessment of bluetongue virus serotype 1 persistence in Spain.

Bluetongue virus (BTV) is an arbovirus of ruminants that has been circulating in Europe continuously for more than two decades and has become endemic in some countries such as Spain. Spain is ideal for BTV epidemiological studies since BTV outbreaks from different sources and serotypes have occurred continuously there since 2000; BTV-1 has been reported there from 2007 to 2017. Here we develop a model for BTV-1 endemic scenario to estimate the risk of an area becoming endemic, as well as to identify the most influential factors for BTV-1 persistence. We created abundance maps at 1-km2 spatial resolution for the main vectors in Spain, Culicoides imicola and Obsoletus and Pulicaris complexes, by combining environmental satellite data with occurrence models and a random forest machine learning algorithm. The endemic model included vector abundance and host-related variables (farm density). The three most relevant variables in the endemic model were the abundance of C. imicola and Obsoletus complex and density of goat farms (AUC 0.86); this model suggests that BTV-1 is more likely to become endemic in central and southwestern regions of Spain. It only requires host- and vector-related variables to identify areas at greater risk of becoming endemic for bluetongue. Our results highlight the importance of suitable Culicoides spp. prediction maps for bluetongue epidemiological studies and decision-making about control and eradication measures.

Large-scale seroprevalence and risk factors associated with Bluetongue virus in Iran.

Bluetongue virus (BTV) remains as an economically major concern in the world. Seroprevalence and potential risk factors of BTV were assessed in a cross-sectional study at both the herd and animal levels in Iran. A total of 73 Epidemiologic Units (E.Unit), defined as a herd, flock or village including animals with equal chance of exposure to infectious agents, were randomly selected. Serum samples from all animals (n = 34,575) within the E.Units were collected and tested for BTV sero-group antibodies by using commercially competitive ELISA test. Using cluster analysis, 90.41 % (95 %, CI: 80.85 %-95.47 %) of the E.Units and 56.13 % (95 % CI: 55.61 %-56.66 %) of the tested animals were detected seropositive against BTV. A seroprevalence rate of 57.59 % (95 % CI: 48.01 %-66.63 %), 65.65 % (95 % CI: 59.10 %-73.74 %) and 27.63 % (95 % CI: 14.40 %-46.43 %) was estimated for sheep, goats and cattle, respectively. At E.Unit (herd) level, density was identified as a great risk factor for the infection (r2 = 0.891; P = 0.000), and particularly density of cattle significantly correlated with BTV infection within the E.Units (r2 = 0.247; P = 0.019). Using multilevel logistic regression, adjusted odds ratios (ORs) were estimated at individual level. A significantly less risk of BTV infection was evaluated for cattle than for sheep (OR = 0.42, 95 % CI: 0.38-0.47, P < 0.001), while no significant difference was observed between sheep and goat (OR = 1.03, 95 % CI: 0.97-1.10, P = 0.345). Animals over 2 years and between 6 months and 2 years expressed 2.22 (OR = 2.22, 95 % CI: 1.96-2.52, P < 0.001) and 2.18 (OR = 2.18, 95 % CI: 1.92-2.49, P < 0.001) times higher chance for the infection than animals under 6 months. Males were at significantly less risk of the infection than females (OR=0.68, 95 % CI: 0.63-0.74, P < 0.001). Animals kept in industrial farming systems displayed 0.46 (OR=0.46, 95 % CI: 032-0.66, P < 0.001) times less chance than animals kept in traditional farming system for BTV, while animals lived in semi-industrial farming system were found to be at 2.97 (OR=2.97, 95 % CI: 2.41-3.66, P < 0.001) times higher chance for BTV than animals lived in traditional farming system. Furthermore, seropositive animals exhibited a high amount of antibodies against BTV (s) suggesting that viral exposure may have frequently occurred during their lifetimes. This large - scale study yielded information on epidemiology of BTV in Iran that is prerequisite for further research, and also for evaluation of any cost-benefit control measure to be established in an enzootic zone of the virus.

Effectiveness and Cost Efficiency of Different Surveillance Components for Proving Freedom and Early Detection of Disease: Bluetongue Serotype 8 in Cattle as Case Study for Belgium, France and the Netherlands.

Quick detection and recovery of country's freedom status remain a constant challenge in animal health surveillance. The efficacy and cost efficiency of different surveillance components in proving the absence of infection or (early) detection of bluetongue serotype 8 in cattle populations within different countries (the Netherlands, France, Belgium) using surveillance data from years 2006 and 2007 were investigated using an adapted scenario tree model approach. First, surveillance components (sentinel, yearly cross-sectional and passive clinical reporting) within each country were evaluated in terms of efficacy for substantiating freedom of infection. Yearly cross-sectional survey and passive clinical reporting performed well within each country with sensitivity of detection values ranging around 0.99. The sentinel component had a sensitivity of detection around 0.7. Secondly, how effective the components were for (early) detection of bluetongue serotype 8 and whether syndromic surveillance on reproductive performance, milk production and mortality data available from the Netherlands and Belgium could be of added value were evaluated. Epidemic curves were used to estimate the timeliness of detection. Sensitivity analysis revealed that expected within-herd prevalence and number of herds processed were the most influential parameters for proving freedom and early detection. Looking at the assumed direct costs, although total costs were low for sentinel and passive clinical surveillance components, passive clinical surveillance together with syndromic surveillance (based on reproductive performance data) turned out most cost-efficient for the detection of bluetongue serotype 8. To conclude, for emerging or re-emerging vectorborne disease that behaves such as bluetongue serotype 8, it is recommended to use passive clinical and syndromic surveillance as early detection systems for maximum cost efficiency and sensitivity. Once an infection is detected and eradicated, cross-sectional screening for substantiating freedom of infection and sentinel for monitoring the disease evolution are recommended.

European Bluetongue Serotype 8: Disease Threat Assessment for U.S. Sheep.

Bluetongue virus (BTV) is an orbivirus transmitted by biting midges (Culicoides spp.) that can result in moderate to high morbidity and mortality primarily in sheep and white-tailed deer. Although only 5 serotypes of BTV are considered endemic to the United States, as many as 11 incursive serotypes have been detected in livestock and wildlife in the past 16 years. Introductions of serotypes, with unknown virulence and disease risk, are constant threats to US agriculture. One potential incursive serotype of particular concern is the European strain of BTV-8, which was introduced into Northern Europe in 2006 and caused unprecedented livestock disease and mortality during the 2006-2007 vector seasons. To assess disease risk of BTV-8 in a common white-faced American sheep breed, eight Polled Dorset yearlings were experimentally infected and monitored for clinical signs. Viremia and viral tissue distribution were detected and quantified by real-time qRT-PCR. Overall, clinical disease was moderate with no mortality. Viremia reached as high as 9.7 log10 particles/mL and persisted at 5 logs or higher through the end of the study (28 days). Virus distribution in tissues was extensive with the highest mean titers at the peak of viremia (day 8) in the kidney (8.38 log10 particles/mg) and pancreas (8.37 log10 particles/mg). Virus persisted in tissues of some sheep at 8 logs or higher by day 28. Results of this study suggest that should BTV-8 emerge in the United States, clinical disease in this common sheep breed would likely be similar in form, duration, and severity to what is typically observed in severe outbreaks of endemic serotypes, not the extraordinary disease levels seen in Northern Europe. In addition, a majority of exposed sheep would be expected to survive and act as significant BTV-8 reservoirs with high titer viremias for subsequent transmission to other livestock and wildlife populations.

Cost analysis of bluetongue virus serotype 8 surveillance and vaccination programmes in Austria from 2005 to 2013.

This study was designed to evaluate the costs between 2005 and 2013 of the national bluetongue virus (BTV) surveillance and vaccination programmes before, during and after the BTV serotype 8 (BTV-8) outbreak in Austria commencing in 2008. In addition to an assessment of the temporal development of costs, a spatial cost analysis was performed. Within the context of this study, the term 'costs' refers to actual financial expenditure and imputed monetary costs for contributions in-kind. Costs were financed directly by the private-public sectors, by the European Commission (EC), and (in-kind) by responsible national institutions and individuals (e.g. blood sampling by veterinarians). The total net cost of the BTV-8 surveillance and vaccination programmes arising from the outbreak amounted to €22.8 million (0.86% of the national agricultural Gross Value Added), of which 32% was allocated to surveillance and 68% to the vaccination programme. Of the total programme costs, the EC supplied €4.9 million, while the remaining costs (€18 million) were directly financed from national resources. Of the latter, €14.5 million was classed as public costs, including €2 million contributions in-kind, and €3.4 million as private costs. The assessment of the costs revealed heterogeneous temporal and spatial distributions. The methodology of this analysis might assist decision makers in calculating costs for other surveillance and intervention programmes. The assessment of contributions in-kind is of importance to public authorities as it increases visibility of the available resources and shows how they have been employed. This study also demonstrates the importance of tracking changing costs per payer over time.

Cost assessment of the movement restriction policy in France during the 2006 bluetongue virus episode (BTV-8).

This study aims at evaluating the costs of the movement restriction policy (MRP) during the 2006 BTV-8 epidemic in France for the producers of 6-9 month old Charolais beef weaned calves (BWC), an important sector that was severely affected by the restrictions imposed. This study estimates the change in the number of BWC sold that was due to the movement restrictions, and evaluates the economic effect of the MRP. The change in BWC sold by producers located inside the restriction zone (RZ) was analyzed for 2006 by using a multivariate matching approach to control for any internal validity threat. The economic evaluation of the MRP was based on several scenarios that describe farms' capacity constraints, feeding prices, and the animal's selling price. Results show that the average farmer experienced a 21% decrease in animals sold due to the MRP. The economic evaluation of the MRP shows a potential gain during the movement standstill period in the case of no capacity constraint faced by the farm and food self-sufficiency. This gain remains limited and close to zero in case of a low selling price and when animals are held until they no longer fit the BWC market so that they cannot be sold as an intermediate product. Capacity constraints represent a tremendous challenge to farmers facing movement restrictions and the fattening profit becomes negative under such conditions. The timing and length of the movement standstill period significantly affect the profitability of the strategy employed by the farmer: for a 5.5 month-long standstill period with 3.5 months of cold weather, farmers with capacity constraints have stronger incentives to leave their animals outside during the whole period and face higher mortality and morbidity rates than paying for a boarding facility for the cold months. This is not necessarily true for a shorter standstill period. Strategies are also sensitive to the feed costs and to the food self-sufficiency of the farm. Altogether, the present work shows the farmer's vulnerability to animal movement restrictions and quantifies the costs of the standstill. These results should assist decision-makers who seek to calculate adequate subsidies/aid or to efficiently allocate resources to prevent future outbreaks.

Expected utility of voluntary vaccination in the middle of an emergent Bluetongue virus serotype 8 epidemic: a decision analysis parameterized for Dutch circumstances.

In order to put a halt to the Bluetongue virus serotype 8 (BTV-8) epidemic in 2008, the European Commission promoted vaccination at a transnational level as a new measure to combat BTV-8. Most European member states opted for a mandatory vaccination campaign, whereas the Netherlands, amongst others, opted for a voluntary campaign. For the latter to be effective, the farmer's willingness to vaccinate should be high enough to reach satisfactory vaccination coverage to stop the spread of the disease. This study looked at a farmer's expected utility of vaccination, which is expected to have a positive impact on the willingness to vaccinate. Decision analysis was used to structure the vaccination decision problem into decisions, events and payoffs, and to define the relationships among these elements. Two scenarios were formulated to distinguish farmers' mindsets, based on differences in dairy heifer management. For each of the scenarios, a decision tree was run for two years to study vaccination behaviour over time. The analysis was done based on the expected utility criterion. This allows to account for the effect of a farmer's risk preference on the vaccination decision. Probabilities were estimated by experts, payoffs were based on an earlier published study. According to the results of the simulation, the farmer decided initially to vaccinate against BTV-8 as the net expected utility of vaccination was positive. Re-vaccination was uncertain due to less expected costs of a continued outbreak. A risk averse farmer in this respect is more likely to re-vaccinate. When heifers were retained for export on the farm, the net expected utility of vaccination was found to be generally larger and thus was re-vaccination more likely to happen. For future animal health programmes that rely on a voluntary approach, results show that the provision of financial incentives can be adjusted to the farmers' willingness to vaccinate over time. Important in this respect are the decision moment and the characteristics of the disease. Farmers' perceptions of the disease risk and about the efficacy of available control options cannot be neglected.

Bluetongue disease risk assessment based on observed and projected Culicoides obsoletus spp. vector densities.

Bluetongue is an arboviral disease of ruminants causing significant economic losses. Our risk assessment is based on the epidemiological key parameter, the basic reproduction number. It is defined as the number of secondary cases caused by one primary case in a fully susceptible host population, in which values greater than one indicate the possibility, i.e., the risk, for a major disease outbreak. In the course of the Bluetongue virus serotype 8 (BTV-8) outbreak in Europe in 2006 we developed such a risk assessment for the University of Veterinary Medicine Vienna, Austria. Basic reproduction numbers were calculated using a well-known formula for vector-borne diseases considering the population densities of hosts (cattle and small ruminants) and vectors (biting midges of the Culicoides obsoletus spp.) as well as temperature dependent rates. The latter comprise the biting and mortality rate of midges as well as the reciprocal of the extrinsic incubation period. Most important, but generally unknown, is the spatio-temporal distribution of the vector density. Therefore, we established a continuously operating daily monitoring to quantify the seasonal cycle of the vector population by a statistical model. We used cross-correlation maps and Poisson regression to describe vector densities by environmental temperature and precipitation. Our results comprise time series of observed and simulated Culicoides obsoletus spp. counts as well as basic reproduction numbers for the period 2009-2011. For a spatio-temporal risk assessment we projected our results from the location of Vienna to the entire region of Austria. We compiled both daily maps of vector densities and the basic reproduction numbers, respectively. Basic reproduction numbers above one were generally found between June and August except in the mountainous regions of the Alps. The highest values coincide with the locations of confirmed BTV cases.

Assessment of the risk of a bluetongue outbreak in Europe caused by Culicoides midges introduced through intracontinental transport and trade networks.

The importation of infected hosts and the arrival of windborne infected Culicoides (Diptera: Ceratopogonidae) were considered unlikely mechanisms for bluetongue virus (BTV) incursion into a BTV-free area during the recent BTV serotype 8 (BTV-8) epidemic in northern Europe. Therefore, alternative mechanisms need to be considered. Air, sea and land transport networks continue to expand, and an important consequence of this is vector-borne pathogen importation. One important aspect of bluetongue (BT) epidemiology not yet addressed is the potential movement of infected Culicoides via transport and trade networks. Therefore, a risk assessment model was constructed to assess the probability of a BTV outbreak as a consequence of the introduction of Culicoides via these networks. The model was applied to calculate the risk for a BTV-8 epidemic in Spain in 2007 caused by the introduction of Culicoides from affected northern European countries. The mean weighted annual risk for an outbreak caused by transportation of a single vector from an affected northern European country varied from 1.8 × 10(-7) to 3.0 × 10(-13), with the highest risks associated with Culicoides imported from Belgium, the Netherlands, Germany and France. For this mechanism to pose a significant risk to BTV-free countries, a large number of vectors would have to be transported.

A two year BTV-8 vaccination follow up: molecular diagnostics and assessment of humoral and cellular immune reactions.

The compulsory vaccination campaign against Bluetongue virus serotype eight (BTV-8) in Germany was exercised in the state of Bavaria using three commercial monovalent inactivated vaccines given provisional marketing authorisation for emergency use. In eleven Bavarian farms representing a cross sectional area of the state the immune reactions of sheep and cattle were followed over a two year period (2008-2009) using cELISA, a serum neutralisation test (SNT) and interferon gamma (IFN-γ) ELISPOT. For molecular diagnostics of BTV genome presence two recommended real time quantitative RT-PCR protocols were applied. The recommended vaccination scheme led to low or even undetectable antibody titers (ELISA) in serum samples of both cattle and sheep. A fourfold increase of the vaccine dose in cattle, however, induced higher ELISA titers and virus neutralising antibodies. Accordingly, repeated vaccination in sheep caused an increase in ELISA-antibody titers. BTV-8 neutralising antibodies occurred in most animals only after multiple vaccinations in the second year of the campaign. The secretion of interferon gamma (IFN-γ) in ELISPOT after in vitro re-stimulation of PBMC of BTV-8 vaccinated animals with BTV was evaluated in the field for the first time. Sera of BTV-8 infected or vaccinated animals neutralising BTV-8 could also neutralise an Italian BTV serotype 1 cell culture adapted strain and PBMC of such animals secreted IFN-γ when stimulated with BTV-1.

Economic evaluation of the surveillance and intervention programme for bluetongue virus serotype 8 in Switzerland.

Empirical analyses founded on sound economic principles are essential in advising policy makers on the efficiency of resource use for disease mitigation. Surveillance and intervention are resource-using activities directed at mitigation. Surveillance helps to offset negative disease effects by promoting successful intervention. Intervention is the process of implementing measures (e.g. vaccination or medication) to reduce or remove a hazard in a population. The scale and ratios in which the two are combined affect the efficiency of mitigation, its costs, benefits, and thus net effect on society's well-being. The Swiss national mitigation programme for bluetongue virus serotype 8 was used as case study to investigate the economic efficiency of mitigation. In 2008, Switzerland implemented a vaccination programme to avoid and reduce disease and infection in its ruminant population. To monitor the vaccination programme and the vector dynamics, a surveillance system consisting of serological and entomological surveillance was established. Retrospective analyses for the years 2008-2009 and prospective analyses for the years 2010-2012 were conducted to investigate if the mitigation programme was economically beneficial. In the retrospective analysis, the implemented programme (=comparative scenario) was compared to a hypothesised baseline scenario of voluntary vaccination and surveillance. In the prospective analysis, the comparative scenario assumed to continue was compared to two baseline scenarios: one of voluntary vaccination combined with surveillance and one of no vaccination combined with surveillance. For each scenario, monetary surveillance, intervention and disease costs were calculated. The comparison of baseline and comparative scenarios yielded estimates for the total benefit (=disease costs avoided), margin over intervention cost and the net value of the programme. For 2008-2009, in aggregate, the mean biannual total benefit was 17.46 m Swiss francs (CHF) (1CHF=0.66€ at the time of analysis) and the mean net benefit after subtraction of the intervention and surveillance cost was 3.95 m CHF. For the three years 2010-2012, overall net costs were estimated at 12.93 m and 8.11 m CHF, respectively, for comparison of the implemented mitigation programme with the two baseline scenarios. It was concluded that the surveillance and intervention programme implemented in 2008-2009 was economically beneficial, while its continuation in the same form in 2010-2012 would produce net costs. These costs were due to the mean intervention cost remaining constant at a level of approximately 11 m CHF per year while the mean total benefit would be gradually reduced in 2010-2012 due to the reduced occurrence of disease in a fully vaccinated population.

Quantitative assessment of the probability of bluetongue virus overwintering by horizontal transmission: application to Germany.

Even though bluetongue virus (BTV) transmission is apparently interrupted during winter, bluetongue outbreaks often reappear in the next season (overwintering). Several mechanisms for BTV overwintering have been proposed, but to date, their relative importance remain unclear. In order to assess the probability of BTV overwintering by persistence in adult vectors, ruminants (through prolonged viraemia) or a combination of both, a quantitative risk assessment model was developed. Furthermore, the model allowed the role played by the residual number of vectors present during winter to be examined, and the effect of a proportion of Culicoides living inside buildings (endophilic behaviour) to be explored. The model was then applied to a real scenario: overwintering in Germany between 2006 and 2007. The results showed that the limited number of vectors active during winter seemed to allow the transmission of BTV during this period, and that while transmission was favoured by the endophilic behaviour of some Culicoides, its effect was limited. Even though transmission was possible, the likelihood of BTV overwintering by the mechanisms studied seemed too low to explain the observed re-emergence of the disease. Therefore, other overwintering mechanisms not considered in the model are likely to have played a significant role in BTV overwintering in Germany between 2006 and 2007.

A broad assessment of factors determining Culicoides imicola abundance: modelling the present and forecasting its future in climate change scenarios.

Bluetongue (BT) is still present in Europe and the introduction of new serotypes from endemic areas in the African continent is a possible threat. Culicoides imicola remains one of the most relevant BT vectors in Spain and research on the environmental determinants driving its life cycle is key to preventing and controlling BT. Our aim was to improve our understanding of the biotic and abiotic determinants of C. imicola by modelling its present abundance, studying the spatial pattern of predicted abundance in relation to BT outbreaks, and investigating how the predicted current distribution and abundance patterns might change under future (2011-2040) scenarios of climate change according to the Intergovernmental Panel on Climate Change. C. imicola abundance data from the bluetongue national surveillance programme were modelled with spatial, topoclimatic, host and soil factors. The influence of these factors was further assessed by variation partitioning procedures. The predicted abundance of C. imicola was also projected to a future period. Variation partitioning demonstrated that the pure effect of host and topoclimate factors explained a high percentage (>80%) of the variation. The pure effect of soil followed in importance in explaining the abundance of C. imicola. A close link was confirmed between C. imicola abundance and BT outbreaks. To the best of our knowledge, this study is the first to consider wild and domestic hosts in predictive modelling for an arthropod vector. The main findings regarding the near future show that there is no evidence to suggest that there will be an important increase in the distribution range of C. imicola; this contrasts with an expected increase in abundance in the areas where it is already present in mainland Spain. What may be expected regarding the future scenario for orbiviruses in mainland Spain, is that higher predicted C. imicola abundance may significantly change the rate of transmission of orbiviruses.

Assessment of bluetongue viraemia in sheep by real-time PCR and correlation with viral infectivity.

Inoculation of embryonated chicken eggs is the standard method for the titration of infectious Bluetongue virus (BTV). Here, six RNA extraction methods coupled with optimised dsRNA denaturation and real-time RT-PCR were evaluated for the quantitation of BTV in blood samples from experimentally infected sheep and results were correlated to infectious virus titres. An exogenous dsRNA internal control (IC) from the closely related Epizootic hemorrhagic disease virus (EHDV) was used to assess the efficiency of BTV genome extraction, dsRNA denaturation, RT, and PCR amplification. Recovery rates of IC and BTV dsRNA copies from extracted blood samples were highly correlated. Adjustment of BTV concentrations according to the IC recovery reduced variation in sample analyses among the different extraction methods and improved the accuracy of BTV quantitation. The EID(50)/ml titre, determined in blood samples from sheep infected experimentally with BTV-1 or BTV-9, correlated highly with the assessed concentration of BTV dsRNA copies. However, this correlation was consistent only during the first 28 days post-infection. The optimised extraction methods and quantitative RT-PCR could be useful for experimental studies of BTV transmission, pathogenesis and vaccine efficacy, or adapted further for the detection and quantitation of EHDV, African horse sickness virus and other dsRNA viruses.

Financial consequences of the Dutch bluetongue serotype 8 epidemics of 2006 and 2007.

This study calculates the financial consequences of the bluetongue serotype 8 (BTV8) epidemics of 2006 and 2007 in the Netherlands. We constructed a deterministic economic model that is compatible with the Dutch livestock production systems for cattle, sheep and goats. Two hundred cattle farms and 270 sheep farms were infected with BTV8 in the epidemic of 2006, whereas 30,417 cattle farms, 45,022 sheep farms and 35,278 goat farms were estimated to be infected in the epidemic of 2007. The net costs (costs minus benefits) of the BTV8 epidemic of 2006 (BT2006) was estimated at 32.4 million Euros. The net costs of the BTV8 epidemic of 2007 (BT2007) was valued at 164-175 million Euros, depending on the mortality and morbidity rates for cattle used. The losses account for 2%, 10% and 11% of the gross value of the primary production within Dutch pasture-based livestock farming that equals 1.6 billion Euros. Control measures accounted for 91% of the net costs of the BT2006, while diagnostic costs represented 7%. By contrast, for the BT2007 92% of the net costs were in the form of production losses and veterinary treatment fees, while only 6% were related to control measures. Furthermore, the control costs dropped from 29,630 in BT2006 to 10,990 in BT2007 mainly due to the costly indoor housing that was not obligatory during the BT2007 epidemic. The cattle sector suffered 88% and 85% of the net costs for the BT2006 and BT2007, respectively; the highest of all sectors.