Technological advances in veterinary diagnostics: opportunities to deploy rapid decentralised tests to detect pathogens affecting livestock.

Sustainable food production capable of feeding a growing human population is a significant global challenge, and is a priority encompassed within the United Nations Millennium Development Goal to 'eradicate extreme poverty and hunger'. Infectious diseases reduce the productivity of farm animals, and the globalised trade of animals and their products increases the threat of disease incursion. Accurate and rapid diagnostic tests are an essential component of contingency plans to detect, control and eradicate such diseases. Diagnosis involves a 'pipeline' that normally starts with clinical suspicion, followed by collecting samples, transporting specimens to a centralised laboratory setting (e.g. national/international Reference Laboratories), analysing these samples using a range of diagnostic tests and reporting the results. However, the transport of specimens from the field to the laboratory can be a lengthy process that can delay critical decision-making and severely affect the quality of the samples. This important limitation of centralised diagnostic testing has motivated the development of tools for the rapid, simple detection of livestock pathogens. Recent advances in the development of technologies for personalised human medicine have motivated the development of prototype diagnostic tests for a wide selection of diseases of livestock. However, many of these tests are not yet routinely used or commercially available. This paper critically reviews the most promising examples of such assays, and highlights the challenges that remain to transition these tests from applied research and development into routine use.

La production durable de denrées alimentaires pour nourrir une population humaine en constante augmentation constitue un vaste enjeu planétaire ainsi que l'une des priorités définies par les Nations Unies dans le cadre des Objectifs du Millénaire pour le développement visant à « éradiquer l'extrême pauvreté et la faim dans le monde ¼. D'une part, les maladies animales réduisent la productivité des animaux d'élevage ; d'autre part, la mondialisation des échanges d'animaux et de produits d'origine animale intensifie les risques d'incursion de maladies. L'utilisation de tests de diagnostic rapides et fiables est une composante essentielle des plans d'urgence visant à détecter, contrôler et éradiquer ces maladies. Une procédure de diagnostic est généralement constituée de plusieurs opérations, depuis la suspicion clinique, la collecte d'échantillons, leur transport vers un laboratoire central (par exemple un laboratoire de référence national/international), jusqu'à l'analyse de ces échantillons au moyen d'une série de tests diagnostiques et la notification des résultats. Néanmoins, le transport des échantillons depuis le terrain jusqu'au laboratoire est parfois un processus très long qui peut retarder la prise de décisions cruciales, voire compromettre gravement la qualité des échantillons. Cette limitation importante des procédures diagnostiques centralisées a incité à mettre au point des outils permettant une détection rapide et aisée des agents pathogènes affectant le bétail. Les progrès récents accomplis dans les technologies relevant de la médecine humaine personnalisée ont encouragé le développement de prototypes d'épreuves de diagnostic pour nombre de maladies du bétail. Toutefois, plusieurs de ces tests ne sont pas encore utilisés en routine ni disponibles commercialement. Les auteurs font le point sur les exemples les plus prometteurs de ces tests et soulignent les difficultés restant à résoudre pour que ces tests puissent évoluer d'une application en recherche et développement à une utilisation en routine.

El logro de una producción sostenible de alimentos en cantidad suficiente para abastecer a una población humana cada vez más numerosa es una difícil empresa que el mundo tiene ante sí, que además entronca con una de las prioridades plasmadas en los Objetivos de Desarrollo del Milenio de las Naciones Unidas: «erradicar la pobreza extrema y el hambre¼. Las enfermedades infecciosas merman la productividad de los animales de granja, al tiempo que el comercio mundializado de animales y sus derivados amplifica la amenaza de incursiones infecciosas. La existencia de pruebas de diagnóstico rápidas y exactas es un elemento básico de todo plan de emergencia encaminado a detectar, controlar y erradicar esas enfermedades. Las labores de diagnóstico entrañan un «circuito¼ que normalmente empieza con la sospecha clínica, sigue con la obtención de muestras, su transporte a un laboratorio central (como un laboratorio de referencia nacional o internacional) y su análisis mediante diversas pruebas de diagnóstico y culmina con la notificación de los resultados. Sin embargo, el transporte hasta un laboratorio de las muestras obtenidas sobre el terreno es a veces un proceso lento, que puede retrasar la adopción de decisiones cruciales y mermar sensiblemente la calidad de las muestras. Este importante inconveniente derivado de la realización centralizada de pruebas ha llevado a concebir herramientas que permitan detectar de forma rápida y sencilla patógenos presentes en el ganado. Los avances registrados últimamente en la obtención de tecnologías destinadas a la medicina humana personalizada han propiciado también la elaboración de prototipos de pruebas para diagnosticar numerosas enfermedades del ganado, aunque muchas de ellas todavía no se utilizan sistemáticamente ni están comercializadas. Los autores, tras examinar en clave crítica los más prometedores ejemplos de estos nuevos ensayos, señalan las dificultades que aún subsisten para que estas pruebas puedan pasar del ámbito de la investigación aplicada y el desarrollo al de su utilización sistemática.

Systematic review of surveillance systems and methods for early detection of exotic, new and re-emerging diseases in animal populations.

In this globalized world, the spread of new, exotic and re-emerging diseases has become one of the most important threats to animal production and public health. This systematic review analyses conventional and novel early detection methods applied to surveillance. In all, 125 scientific documents were considered for this study. Exotic (n = 49) and re-emerging (n = 27) diseases constituted the most frequently represented health threats. In addition, the majority of studies were related to zoonoses (n = 66). The approaches found in the review could be divided in surveillance modalities, both active (n = 23) and passive (n = 5); and tools and methodologies that support surveillance activities (n = 57). Combinations of surveillance modalities and tools (n = 40) were also found. Risk-based approaches were very common (n = 60), especially in the papers describing tools and methodologies (n = 50). The main applications, benefits and limitations of each approach were extracted from the papers. This information will be very useful for informing the development of tools to facilitate the design of cost-effective surveillance strategies. Thus, the current literature review provides key information about the advantages, disadvantages, limitations and potential application of methodologies for the early detection of new, exotic and re-emerging diseases.

New insights into the role of ticks in African swine fever epidemiology.

African swine fever (ASF), one of the most important diseases of swine, is present in many African countries, as well as in eastern Europe, Russia and Sardinia. It is caused by a complex virus, ASF virus (ASFV), for which neither vaccine nor treatment is available. ASFV affects swine of all breeds and ages, and also replicates in soft ticks of the genus Ornithodoros, facilitating ASFV persistence and reocurrence of disease. Depending on the involvement of these ticks, and the presence or not of sylvatic asymptomatic animals, several epidemiological cycles have been identified. The disease persists in East and southern African countries in a sylvatic cycle between O. porcinus (of the O. moubata species complex) and common warthogs. In some countries a domestic pig-tick cycle exists, whereas in other regions, notably West Africa, the role of soft ticks has not been demonstrated, and ASFV is transmitted between domestic pigs in the absence of tick vectors. Even in several East and Central African countries which have the sylvatic or domestic cycle, the majority of outbreaks are not associated with ticks or wild suids. In Europe, O. erraticus was detected and identified as a crucial vector for ASF maintenance in outdoor pig production on the Iberian Peninsula. However, in most parts of Europe, there is a lack of information about the distribution and role of Ornithodoros ticks in ASF persistence, particularly in eastern regions. This article reviews ASF epidemiology and its main characteristics, with a special focus on the distribution and role of soft ticks in ASF persistence in different settings. Information abouttick detection, control measures and future directions for research is also included.

Development and evaluation of a SYBR Green real-time RT-PCR assay for evaluation of cytokine gene expression in horse.

Cytokine secretion is one of the main mechanisms by which the immune system is regulated in response to pathogens. Therefore, the measurement of cytokine expression is fundamental to characterizing the immune response to infections. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) is widely used to measure cytokine mRNA levels, but assay conditions should be properly evaluated before analyzing important equine infections through relative quantification of gene expression. The aim of this study was to develop and evaluate a set of RT-qPCR assays for a panel of the most common cytokines in horses involved in innate and adaptive immune responses. Eight cytokines (interleukin (IL)-1ß, IL-2, IL-4, IL-10, IL-12, TNFα, IFNß and IFNγ) and a housekeeping gene (ß-actin) were detected and amplified with the same annealing temperature in a SYBR Green RT-qPCR assay of samples of mitogen-stimulated peripheral blood mononuclear cells from a healthy horse and whole blood from a horse infected with African horse sickness virus. The method gave good efficiency for all genes tested, allowing quantification of relative expression levels. These SYBR Green RT-qPCR assays may be useful for examining cytokine gene expression in horses in response to exposure to economically important pathogens.

African swine fever diagnosis update.

African swine fever (ASF) is one of the most complex livestock diseases. The significant losses that it causes, coupled with the lack of a vaccine against ASF virus and the possible resemblance with other swine hemorrhagic diseases, make early detection and laboratory diagnosis essential for controlling and managing the disease. All the techniques currently used to diagnose ASF have been fully validated showing high sensitivity and specificity to detect both antigen and antibodies against all 22 known genotypes; and enable the correct diagnosis of ASF in all possible epidemiological situations. Because no vaccine is available, the presence of antibodies always indicates previous infection, and serological diagnosis must always be performed in parallel with antigen detection to increase the sensitivity and specificity of the analyses. Recent developments in ASF diagnosis, specifically the new field diagnostic tests, have improved and facilitated the likelihood of ASF early detection, essential to fighting the disease.

Identification of suitable areas for West Nile virus outbreaks in equid populations for application in surveillance plans: the example of the Castile and Leon region of Spain.

The introduction and rapid spread of West Nile virus (WNV) into new areas such as the American continent, associated also with the severity of the disease in humans and equids has increased concerns regarding the need to better prevent and control future WNV incursions. WNV outbreaks in equids usually occur under specific climatic and environmental conditions and, typically, before detection of WNV cases in humans. Targeting surveillance strategies in areas and time periods identified as suitable for WNV outbreaks in equids may act as an early-warning system to prevent disease in both equids and humans. This study used a GIS-based framework to identify suitable areas and time periods for WNV outbreak occurrence in one of the most important areas of equid production in Spain, i.e. Castile and Leon. Methods and results presented here may help to improve the early detection and control of future WNV outbreaks in Spain and other regions.

[African swine fever in Russian Federation].

African swine fever (ASF) is an infectious viral disease that causes high economic losses due to the necessity of depopulation of pigs in affected areas, sanitary measures, trade restrictions, etc. The virus (ASFV) is relatively stable in the unprocessed meat products and environment. Thus, large areas are at risk due to free movement of people and products. The ASFV does not affect people and animals, except the wild and domestic pigs. Some ticks can become infected and carry the virus for years. Adaptation of the virus by changing into the less virulent form would mean the threat of an endemic situation to the area. The disease is endemic in domestic and wild pigs in most of sub-Saharan Africa and Sardinia, Italy. There is no treatment for ASF, and no vaccine has been developed. In case of infection with less virulent ASFV strains, the recovered pigs could spread the virus as long as their live. In terms of clinical symptoms, ASF is very similar to Classical Swine Fever. The methods of laboratory diagnostics are well developed and efficient for identification of ASFV and virus-specific antibodies. Experience of eradication of ASF in Spain suggests the importance of serological monitoring of pigs. In the spring of 2007, the ASF was detected in the Caucasus region. Same virus was detected in Georgia, Armenia, Azerbaijan, and Russia. The ASFV circulating in the Caucasus and the Russian Federation is a highly virulent virus. No reduction of the virulence was observed since the first outbreak in Georgia. In the last years, the ASF remained in the Caucasus, southern parts of Russia and appeared occasionally as far as St. Petersburg and St. Petersburg region, and in the area of Nizhny Novgorod. Domestic pigs play an important role in the ASFV spread; they transfer the virus to the wild boars. The virus circulates in the population of wild boars depending on their density in the area. Occasionally, the disease is spread from wild to domestic pigs. There is no evidence of ticks being involved in the process. Thus, the human activity in raising pigs is largely responsible for continuous spread of the disease. Despite vigorous monitoring and sanitary measures, the disease has not been stopped. The control strategy for ASF should consider International (especially Spanish) experience and local situation. The strategy is based on the number of important steps including rapid localization of the disease by trained specialists, setting up buffer zones, constant serologic monitoring of swine population and farms, improvement of diagnostic facilities, training of veterinary personnel, development of the system of information and international collaboration.

Reproductive ratio for the local spread of highly pathogenic avian influenza in wild bird populations of Europe, 2005-2008.

Highly pathogenic avian influenza (HPAI) has devastating consequences for the poultry industry of affected countries. Control of HPAI has been impaired by the role of wildlife species that act as disease reservoirs and as a potential source of infection for domestic populations. The reproductive ratio (R0) of HPAI was quantified in nine clusters of outbreaks detected in wild birds in Europe (2005-2008) for which population data were not available. The median value of R0 was similar (1·1-3·4) for the nine clusters and it was about tenfold smaller than the value estimated for poultry in The Netherlands in 2003. Results presented here will be useful to parameterize models for spread of HPAI in wild birds and to design effective prevention programmes for the European poultry sector. The method is suitable to estimate R0 in the absence of population data, which is a condition typically observed for many wildlife and certain domestic species and systems.

Evaluating surveillance in wild birds by the application of risk assessment of avian influenza introduction into Spain.

Early detection of highly pathogenic avian influenza (HPAI) in its natural reservoirs is a prerequisite for preventing disease spread to humans. The risk of introduction of H5N1 HPAI was assessed in order to design a risk-based surveillance system in Spain. Areas at highest risk for H5N1 HPAI followed a northeast-southwest direction, with two significant clusters located in the north and the southwest of the country. Most (83%) of the veterinary units (VUs) obtained fewer samples than would have been expected if samples had been collected using a risk-based design. In October 2009, a HPAI outbreak was reported for the first time in a Spanish layer farm located in a VU at high risk for HPAI, but no samples were collected. This risk-based surveillance approach will increase the cost-effectiveness of HPAI surveillance in Spain and can be easily extended to and adopted by other countries and regions throughout the world.

Bottlenose dolphins (Tursiops truncatus) aggressive behavior towards other cetacean species in the western Mediterranean.

Aggressive behavior of bottlenose dolphins (Tursiops truncatus) towards conspecifics is widely described, but they have also often been reported attacking and killing harbour porpoises (Phocoena phocoena) around the world. However, very few reports exist of aggressive interactions between bottlenose dolphins and other cetacean species. Here, we provide the first evidence that bottlenose dolphins in the western Mediterranean exhibit aggressive behavior towards both striped dolphins (Stenella coeruleoalba) and Risso's dolphins (Grampus griseus). Necropsies and visual examination of stranded striped (14) and Risso's (2) dolphins showed numerous lesions (external rake marks and different bone fractures or internal organ damage by blunt trauma). Indicatively, these lessons matched the inter-tooth distance and features of bottlenose dolphins. In all instances, these traumatic interactions were presumed to be the leading cause of the death. We discuss how habitat changes, dietary shifts, and/or human colonization of marine areas may be promoting these interactions.

A stochastic model to quantify the risk of introduction of classical swine fever virus through import of domestic and wild boars.

Classical swine fever (CSF) is a disease of pigs that imposes major hardship on the industry of infected regions. The recent history of CSF epidemics suggests that animal movements remain the main source of CSF virus (CSFV) infection for susceptible populations in Europe. This study presents an assessment of the risk of introducing CSFV into Spain through the importation of live susceptible animals. Results suggest that, if prevailing conditions persist, introduction of CSFV into Spain is likely to occur on average every 9 years and that introduction is almost three times more likely to occur via domestic pigs than through wild boars. The highest risk was concentrated in March and in the Northeastern provinces of Spain. Results were consistent with the time and location of previous CSFV introductions into the country. The methodology and the results presented here will contribute to improve the CSF prevention programme in Spain.

Rapid and sensitive detection of African horse sickness virus by real-time PCR.

A highly sensitive and specific TaqMan-MGB real-time RT-PCR assay has been developed and standardised for the detection of African horse sickness virus (AHSV). Primers and MGB probe specific for AHSV were selected within a highly conserved region of genome segment 7. The robustness and general application of the diagnostic method were verified by the detection of 12 AHSV isolates from all of the nine serotypes. The analytical sensitivity ranged from 0.001 to 0.15 TCID(50) per reaction, depending on the viral serotype. Real-time PCR performance was preliminarily assessed by analysing a panel of field equine samples. The same primer pair was used to standardise a conventional RT-PCR as an affordable, useful and simple alternative method in laboratories without access to real-time PCR instruments. The two techniques present novel tools to improve the molecular diagnosis of African horse sickness (AHS).

Determination of the immunotoxic potential of heavy metals on the functional activity of bottlenose dolphin leukocytes in vitro.

Heavy metals may affect the immune system of cetaceans. But no information exists on their effects on the bottlenose dolphin (Tursiops truncatus) immune system, although this species is a coastal top predator which can bioaccumulate high concentrations of them. This work studies the effects of Hg (1, 5 and 10mg/L), Al (2,5, 25 and 50mg/L), Cd (1, 10, 20 and 40mg/L), Pb (1, 10, 20 and 50mg/L) and Cr (1 and 10mg/L), on the function of phagocytes and lymphocytes isolated from the peripheral blood of bottlenose dolphins under in vitro conditions. Cell viability, apoptosis, lymphocyte proliferation and phagocytosis were evaluated. Viability and lymphoproliferation were measured with Alamar Blue assay, and apoptosis and phagocytosis were evaluated with flow cytometry. Apoptosis was detected as mechanism of cell death after cadmium and mercury exposure. A significant reduction in the lymphoproliferative response was registered by exposure to 1mg/L of mercury, 10mg/L of cadmium and 50mg/L of lead. Decreased phagocytosis was also observed at 5mg/L of mercury, 50mg/L of aluminium and 10mg/L of cadmium. Chromium did not present any effects on any immune assay at the concentrations tested. The concentrations of heavy metals that were found to affect the functional activity of bottlenose dolphin leukocytes are within the environmental ranges reported in the tissues of bottlenose dolphins. These results support the hypothesis that exposure to these contaminants, particularly mercury and cadmium could lead to a reduction in host resistance to disease in these animals.

Herpes simplex-like infection in a bottlenose dolphin stranded in the Canary Islands.

A bottlenose dolphin, stranded in the Canary Islands in 2001 exhibited non-suppurative encephalitis. No molecular detection of cetacean morbillivirus (CeMV) was found, but a herpesviral-specific band of 250 bp was detected in the lung and brain. The sequenced herpesviral PCR product was compared with GenBank sequences, obtaining 98% homology (p-distance of 0.02) with Human herpesvirus 1 (herpes simplex virus 1 or HSV-1). This is the first report of a herpes simplex-like infection in a stranded dolphin.

Gaps in African swine fever: Analysis and priorities.

African swine fever (ASF) causes greater sanitary, social and economic impacts on swine herds than many other swine diseases. Although ASF was first described in 1921 and it has affected more than fifty countries in Africa, Europe and South America, several key issues about its pathogenesis, immune evasion and epidemiology remain uncertain. This article reviews the main characteristics of the causative virus, its molecular epidemiology, natural hosts, clinical features, epidemiology and control worldwide. It also identifies and prioritizes gaps in ASF from a horizontal point of view encompassing fields including molecular biology, epidemiology, prevention, diagnosis and vaccine development. The purpose of this review is to promote ASF research and enhance its control.

Why is African swine fever still present in Sardinia?

African swine fever (ASF) is an infectious disease of swine that has been present in Sardinia since 1978. Soon after introduction of the disease, several control and eradication programmes were established with limited success. Some researchers attributed the persistence of the disease in central and eastern areas to certain socio-economic factors, the existence of some local and traditional farming practices (i.e., unregistered free-ranging pigs known as brado animals) and the high density of wild boar in the region. In the past, scarcity of swine data in Sardinia complicated the evaluation and study of ASF on the island. More complete, accurate and reliable information on pig farms has become available as a result of the most recent eradication programmes. Here, we perform statistical modelling based on these data and the known distribution of domestic pig and wild boar to identify the main risk factors that have caused ASF persistence in Sardinia. Our results categorized, identified and quantified nine significant risk factors, six of which have not been previously described. The most significant factors were the number of medium-sized farms, the presence of brado animals and the combination of estimated wild boar density and mean altitude above sea level. Based on these factors, we identified regions in eastern and central Sardinia to be at greatest risk of ASF persistence; these regions are also where the disease has traditionally been endemic. Based on these risk factors, we propose specific control measures aimed at mitigating such risks and eradicating ASF from the island.

Understanding African Swine Fever infection dynamics in Sardinia using a spatially explicit transmission model in domestic pig farms.

African swine fever virus (ASFV) has been endemic in Sardinia since 1978, resulting in severe losses for local pig producers and creating important problems for the island's veterinary authorities. This study used a spatially explicit stochastic transmission model followed by two regression models to investigate the dynamics of ASFV spread amongst domestic pig farms, to identify geographic areas at highest risk and determine the role of different susceptible pig populations (registered domestic pigs, non-registered domestic pigs [brado] and wild boar) in ASF occurrence. We simulated transmission within and between farms using an adapted version of the previously described model known as Be-FAST. Results from the model revealed a generally low diffusion of ASF in Sardinia, with only 24% of the simulations resulting in disease spread, and for each simulated outbreak on average only four farms and 66 pigs were affected. Overall, local spread (indirect transmission between farms within a 2 km radius through fomites) was the most common route of transmission, being responsible for 98.6% of secondary cases. The risk of ASF occurrence for each domestic pig farm was estimated from the spread model results and integrated in two regression models together with available data for brado and wild boar populations. There was a significant association between the density of all three populations (domestic pigs, brado, and wild boar) and ASF occurrence in Sardinia. The most significant risk factors were the high densities of brado (OR = 2.2) and wild boar (OR = 2.1). The results of both analyses demonstrated that ASF epidemiology and infection dynamics in Sardinia create a complex and multifactorial disease situation, where all susceptible populations play an important role. To stop ASF transmission in Sardinia, three main factors (improving biosecurity on domestic pig farms, eliminating brado practices and better management of wild boars) need to be addressed.

Risk assessment applied to Spain's prevention strategy against highly pathogenic avian influenza virus H5N1.

Notifiable avian influenza (NAI) had never been reported in Spain, until July 2006 when a dead Great Crested Grebe (Podiceps cristatus) was found positive to the highly pathogenic H5N1 subtype as part of the active wild bird surveillance plan. The current program of the Spanish Ministry of Agriculture, Fisheries, and Food (MAPA)'s strategic preventive plan against NAI is divided in the following parts: identification of risk areas and risk wild bird species, increased biosecurity measures, early detection of infection with surveillance intensification and development of rapid diagnostic tests, and other policies, which include continuing education and training to ensure early detection of the disease. In 2003 an active surveillance plan was introduced for domestic fowl; the plan was extended to wild birds in 2004. A total of 18,780 samples in poultry and 3687 samples in wild birds had been analyzed through December 2005 to detect the presence and spread of avian influenza subtypes H5 and H7. In the present work we suggest some contributions to be implemented in MAPA's action plan: 1) the identification of risks because of migratory birds, within the risk assessment of the introduction of NAI virus in Spain and 2) an interactive digital simulator of the disease developed for continuing education and training.

Early Detection of Infection in Pigs through an Online Monitoring System.

Late detection of emergency diseases causes significant economic losses for pig producers and governments. As the first signs of animal infection are usually fever and reduced motion that lead to reduced consumption of water and feed, we developed a novel smart system to monitor body temperature and motion in real time, facilitating the early detection of infectious diseases. In this study, carried out within the framework of the European Union research project Rapidia Field, we tested the smart system on 10 pigs experimentally infected with two doses of an attenuated strain of African swine fever. Biosensors and an accelerometer embedded in an eartag captured data before and after infection, and video cameras were used to monitor the animals 24 h per day. The results showed that in 8 of 9 cases, the monitoring system detected infection onset as an increase in body temperature and decrease in movement before or simultaneously with fever detection based on rectal temperature measurement, observation of clinical signs, the decrease in water consumption or positive qPCR detection of virus. In addition, this decrease in movement was reliably detected using automatic analysis of video images therefore providing an inexpensive alternative to direct motion measurement. The system can be set up to alert staff when high fever, reduced motion or both are detected in one or more animals. This system may be useful for monitoring sentinel herds in real time, considerably reducing the financial and logistical costs of periodic sampling and increasing the chances of early detection of infection.

Effectiveness and practicality of control strategies for African swine fever: what do we really know?

African swine fever (ASF) is a major pig health problem, and the causative virus is moving closer to Western European regions where pig density is high. Stopping or slowing down the spread of ASF requires mitigation strategies that are both effective and practical. Based on the elicitation of ASF expert opinion, this study identified surveillance and intervention strategies for ASF that are perceived as the most effective by providing the best combination between effectiveness and practicality. Among the 20 surveillance strategies that were identified, passive surveillance of wild boar and syndromic surveillance of pig mortality were considered to be the most effective surveillance strategies for controlling ASF virus spread. Among the 22 intervention strategies that were identified, culling of all infected herds and movement bans for neighbouring herds were regarded as the most effective intervention strategies. Active surveillance and carcase removal in wild boar populations were rated as the most effective surveillance and intervention strategies, but were also considered to be the least practical, suggesting that more research is needed to develop more effective methods for controlling ASF in wild boar populations.