The Effectiveness of Protection and Surveillance Zones in Detecting Further African Swine Fever Outbreaks in Domestic Pigs-Experience of the Baltic States.

In the event of an outbreak of African swine fever (ASF) in pig farms, the European Union (EU) legislation requires the establishment of a restricted zone, consisting of a protection zone with a radius of at least 3 km and a surveillance zone with a radius of at least 10 km around the outbreak. The main purpose of the restricted zone is to stop the spread of the disease by detecting further outbreaks. We evaluated the effectiveness and necessity of the restricted zone in the Baltic States by looking at how many secondary outbreaks were detected inside and outside the protection and surveillance zones and by what means. Secondary outbreaks are outbreaks with an epidemiological link to a primary outbreak while a primary outbreak is an outbreak that is not epidemiologically linked to any previous outbreak. From 2014 to 2023, a total of 272 outbreaks in domestic pigs were confirmed, where 263 (96.7%) were primary outbreaks and 9 (3.3%) were secondary outbreaks. Eight of the secondary outbreaks were detected by epidemiological enquiry and one by passive surveillance. Epidemiological enquiries are legally required investigations on an outbreak farm to find out when and how the virus entered the farm and to obtain information on contact farms where the ASF virus may have been spread. Of the eight secondary outbreaks detected by epidemiological investigations, six were within the protection zone, one was within the surveillance zone and one outside the restricted zone. Epidemiological investigations were therefore the most effective means of detecting secondary outbreaks, whether inside or outside the restricted zones, while active surveillance was not effective. Active surveillance are legally prescribed activities carried out by the competent authorities in the restricted zones. Furthermore, as ASF is no longer a rare and exotic disease in the EU, it could be listed as a "Category B" disease, which in turn would allow for more flexibility and "tailor-made" control measures, e.g., regarding the size of the restricted zone.

Identification of predilection sites for wild boar carcass search based on spatial analysis of Latvian ASF surveillance data.

Targeted search for wild boar carcasses is essential for successful control of African swine fever (ASF) in wild boar populations. To examine whether landscape conditions influence the probability of finding ASF-positive carcasses, this study analyzed Global Positioning System (GPS) coordinates of Latvian wild boar carcasses and hunted wild boar, extracted from the CSF/ASF wild boar surveillance database of the European Union, and random coordinates in Latvia. Geographic information system (GIS) software was used to determine the landscape type and landscape composition of carcass detection sites and to measure distances from the carcasses to nearest waterbodies, forest edges, roads and settlements. The results of the automated measurements were validated by manually analyzing a smaller sample. Wild boar carcasses were found predominantly in forested areas and closer to waterbodies and forest edges than random GPS coordinates in Latvia. Carcasses of ASF-infected wild boar were found more frequently in transitional zones between forest and woodland shrub, and at greater distances from roads and settlements compared to ASF-negative carcasses and random points. This leads to the hypothesis, that ASF-infected animals seek shelter in quiet areas further away from human disturbance. A detailed collection of information on the environment surrounding carcass detection sites is needed to characterize predilection sites more accurately.

Retrospective analysis of sero-prevalence and bait uptake estimations in foxes after oral rabies vaccination programmes at European level: Lessons learned and paths forward.

Rabies caused by the Classical Rabies Virus (Lyssavirus rabies abbreviated RABV) in the European Union has been close to elimination mainly thanks to Oral Rabies Vaccination (ORV) campaigns targeting wildlife (primarily red foxes). ORV programmes co-financed by the European Commission include a monitoring-component to assess the effectiveness of the ORV campaigns at national level. This assessment is performed by a random collection of red foxes in the vaccinated areas with control of antibodies presence by serological analysis and control of bait uptake by detection of biomarkers (tetracycline incorporated into the baits) in the bones and teeth. ORV programmes aim to a vaccine coverage high enough to immunize (ideally) 70 % of the reservoir population to control the spread of the disease. European Union (EU) programmes that led to almost elimination of rabies on the territory have been traditionally found to have a bait uptake average of 70 % (EU countries; 2010-2020 period) while the seroconversion data showed an average level of 40 % (EU countries; 2010-2020 period). To better understand variations of these indicators, a study was been set up to evaluate the impact of several variables (linked to the vaccination programme itself and linked to environmental conditions) on the bait uptake and the seroconversion rate. Thus, pooling data from several countries provides more powerful statistics and the highest probability of detecting trends. Results of this study advocate the use of a single serological test across the EU since data variation due to the type of test used was higher than variations due to field factors, making the interpretation of monitoring results at EU level challenging. In addition, the results indicates a negative correlation between bait uptake and maximum temperatures reached during ORV campaigns questioning the potential impact of climatic change and associated increase of temperatures on the ORV programmes efficiency. Several hypotheses requesting additional investigation are drawn and discussed in this paper.

Do Seropositive Wild Boars Pose a Risk for the Spread of African Swine Fever? Analysis of Field Data from Latvia and Lithuania.

In 2020, ASF occurred in wild boars throughout Latvia and Lithuania, and more than 21,500 animals were hunted and tested for the presence of the virus genome and antibodies in the framework of routine disease surveillance. The aim of our study was to re-examine hunted wild boars that tested positive for the antibodies and negative for the virus genome in the blood (n = 244) and to see if the virus genome can still be found in the bone marrow, as an indicator of virus persistence in the animal. Via this approach, we intended to answer the question of whether seropositive animals play a role in the spread of the disease. In total, 2 seropositive animals out of 244 were found to be positive for the ASF virus genome in the bone marrow. The results indicate that seropositive animals, which theoretically could also be virus shedders, can hardly be found in the field and thus do not play an epidemiological role regarding virus perpetuation, at least not in the wild boar populations we studied.

The First Eighteen Months of African Swine Fever in Wild Boar in Saxony, Germany and Latvia-A Comparison.

African swine fever (ASF) emerged in Latvia in 2014. In 2020, the virus has been detected in the German federal state, Saxony. In both regions, the virus was probably introduced by infected wild boar coming from affected neighboring countries. As the current ASF control strategy at EU level had not yet been developed at the time of ASF introduction into Latvia, disease control measures in both study areas differed over time. Assessing the course of ASF in Saxony and the implemented control strategies, the first 18 months of the ASF epidemic were epidemiologically compared between Saxony and Latvia. ASF wild boar surveillance data were analyzed and the prevalence of ASF virus-positive wild boar was estimated. For estimating the wild boar density, the numbers of wild boar per km² were calculated for the respective geographical areas. The number of samples collected from hunted wild boar and wild boar found dead was higher in Saxony. The ASF virus prevalence in Latvia was significantly higher than in Saxony, indicating that Saxony has had more time for getting prepared for dealing with an ASF incursion. Experience from other countries and the rapid implementation of new control strategies may have helped Saxony deal with ASF.

African Swine Fever Outbreak Investigations-The Significance of Disease-Related Anecdotal Information Coming from Laypersons.

Veterinarians who have conducted numerous investigations of African swine fever outbreaks in pig farms in various European countries over the years shared their experiences during a workshop in Germany in early 2020. One focus was on the so-called "anecdotal information" obtained from farmers, farm workers or other lay people during the outbreak investigations. Discussions revolved around how to correctly interpret and classify such information and how the subjective character of the statements can influence follow-up examinations. The statements of the lay persons were grouped into three categories according to their plausibility: (i) statements that were plausible and prompted further investigation, (ii) statements that were not plausible and could therefore be ignored, and (iii) statements that were rather implausible but should not be ignored completely. The easiest to deal with were statements that could be classified without doubt as important and very plausible and statements that were not plausible at all. Particularly difficult to assess were statements that had a certain plausibility and could not be immediately dismissed out of hand. We aim to show that during outbreak investigations, one is confronted with human subjective stories that are difficult to interpret but still important to understand the overall picture. Here, we present and briefly discuss an arbitrary selection of reports made by lay persons during outbreak investigations.

Eight Years of African Swine Fever in the Baltic States: Epidemiological Reflections.

African swine fever (ASF) was first detected in Lithuania, Latvia, and Estonia in 2014 and has since been circulating in the Baltic States with a similar epidemiological course characterized by persistence of the disease in the wild boar population and occasional spill-over infections in domestic pigs. The aim of the present study was to evaluate surveillance data on ASF in wild boar from the three countries to improve our understanding of the course of the disease. ASF surveillance and wild boar population data of the countries were analyzed. In all three countries, a decrease in the prevalence of ASF virus-positive wild boar was observed over time. Although somewhat delayed, an increase in the seroprevalence was seen. At the same time, the wild boar population density decreased significantly. Towards the end of the study period, the wild boar population recovered, and the prevalence of ASF virus-positive wild boar increased again, whereas the seroprevalence decreased. The decreasing virus prevalence has obviously led to virus circulation at a very low level. Together with the decreasing wild boar population density, the detection of ASF-infected wild boar and thus ASF control has become increasingly difficult. The course of ASF and its continuous spread clearly demonstrate the necessity to scrutinize current ASF surveillance and control strategies fundamentally and to consider new transdisciplinary approaches.

A practical guide for strategic and efficient sampling in African swine fever-affected pig farms.

In the case of African swine fever (ASF) outbreaks in pig farms, EU legislation requires a thorough epidemiological investigation to determine, among other tasks, the extent of infection in the affected farm. The main aim of this study was to implement a reliable sampling strategy to quickly obtain an overview of the extent of ASF virus spread in an affected pig farm. We developed and tested a three-step approach: (i) identification of sub-units within the affected farm, (ii) categorization of sub-units, and (iii) targeted selection of animals for testing. We used commercially available lateral flow devices (LFDs) to detect ASF antigen and antibodies under field conditions and compared them with routinely performed laboratory tests (qPCR, ELISA, IPT). The study was conducted in three commercial farms in Latvia that were affected by ASF in July 2020. One of the affected farms was relatively small with only 31 pigs, whereas the other two were large with 1800 and 9800 animals, respectively. The approach proved to be helpful and practical for efficient and reliably assess the ASF situation on the farm and to identify sub-units within a farm where infected animals are present and sub-units which might (still) be free of infection. This important epidemiological information helps to better estimate the high-risk period and to track the potential spread of infection outside the farm. It allows also to prioritize culling and, if appropriate, to pursue a partial culling strategy taking into account the absence of clinical signs, implemented biosecurity measures, quarantine and negative test results, among others. This might be of interest for large commercial farms where the infection was identified very early and has not yet spread widely. Due to its limited sensitivity, the antigen LFD test is useful for testing animals showing signs of disease.

Hunters' view on the control of African swine fever in wild boar. A participatory study in Latvia.

African swine fever (ASF) has massively spread in recent years and threatens the global pig industry. ASF has been present in Latvia since 2014. Hunters play a major role in the implementation of measures to control ASF and in passive disease surveillance. The probability to detect an ASF-positive wild boar is much higher in animals found dead than in hunted animals. Thus, the willingness and the motivation of hunters to support passive surveillance is of utmost importance. Using participatory methods, this study aimed to assess the acceptability of control measures for ASF in wild boar among hunters. In addition, new approaches to increase hunters' motivation to report wild boar found dead were investigated. A total of ten focus group discussions with hunters from different regions in Latvia were conducted. To assess the quantity and quality of contacts between hunters and stakeholders involved in the control of ASF, relation diagrams were used. Using ranking tools, the trust of the participants in stakeholders to implement control measures successfully was evaluated. Defined control measures were presented to the hunters and their acceptability investigated. An impact diagram and a list of defined motivation options for passive surveillance were offered to identify new ways to increase the willingness of hunters to support passive surveillance actively. A satisfactory and regular relationship was identified between the hunters, the Food and Veterinary Service (FVS) and the State Forest Service (SFS). The hunters' trust in these authorities was high. Although there is no vaccine against ASF, hunters were convinced of the potential of vaccination in controlling ASF. However, building fences was considered as useless and ineffective. To increase the willingness of hunters to support passive surveillance, reducing the infection pressure in the forests was regarded as most motivating. Furthermore, hunters would appreciate a decrease in their costs and workload. The study provides new insight into the concerns and experiences of hunters. Including their views and expectations in the further design and implementation of control and surveillance activities may help to improve current efforts to control ASF in wild boar populations. Although representing the perceptions of Latvian hunters, the main conclusions may be adaptable to adjust ASF control and surveillance in other countries.

African Swine Fever in Two Large Commercial Pig Farms in LATVIA-Estimation of the High Risk Period and Virus Spread within the Farm.

African swine fever (ASF) was first detected in Latvia in wild boar at the Eastern border in June 2014. Since then ASF has continued to spread in wild boar populations covering almost whole territory of the country. Sporadic outbreaks occurred at the same time in domestic pig holdings located in wild boar infected areas. Here we present the results of the epidemiological investigation in two large commercial farms. Several parameters were analyzed to determine the high risk period (HRP) and to investigate the ASF virus spread within the farm. Clinical data, mortality rates and laboratory results proved to be good indicators for estimating the HRP. The measures for early disease detection, particularly the enhanced passive surveillance that is targeting dead and sick pigs, were analyzed and discussed. Enhanced passive surveillance proved to be a key element to detect ASF at an early stage. The study also showed that ASF virus might spread slowly within a large farm depending mainly on direct contacts between pigs and the level of internal biosecurity. Findings suggest improvements in outbreak prevention, control measures and may contribute to a better understanding of ASF spreading patterns within large pig herds. Culling of all pigs in large commercial farms could be reconsidered under certain conditions.

African swine fever in Latvian wild boar-A step closer to elimination.

In 2014, African swine fever (ASF) emerged in Latvia for the first time. The majority of cases appeared in wild boar, but the presence of ASF in these animals constitutes a permanent threat to domestic pig holdings. Recent studies have shown an increase in serologically positive and a decrease in PCR-positive ASF cases in wild boar, possibly indicating a decline of ASF incidence. We aimed to investigate the course of the ASF epidemic in wild boar in Latvia, thus attaining further insights into the ASF epidemiology in this country with the goal of assessing the stage of the epidemic. Latvian ASF surveillance data of wild boar were utilized to estimate the seroprevalence and ASF virus (ASFV) prevalence in the wild boar population. Prevalence estimates were obtained for both the eastern and western part of the country and in addition for the 2014/2015 to 2018/2019 hunting seasons. Moreover, prevalence estimates for three different age classes were calculated. An increase in serologically positive yet PCR-negative wild boar samples from active surveillance was identified over time. When comparing the age groups, wild boar younger than one year displayed the ASFV prevalence to be higher than the seroprevalence, whereas older animals shared higher seroprevalence estimates. These findings support the assumption that only a small proportion of affected animals survive an infection, leading to an accumulation of their numbers over time. As a result, ASF elimination in a country with an infected wild boar population could possibly be achieved, if effective wild boar population management and surveillance is maintained and combined with the detection and removal of wild boar carcasses to reduce the viral load in the environment. In addition, the wild boar population should be kept as small as possible to break the ASFV infection cycle.

Epidemiological evaluation of Latvian control measures for African swine fever in wild boar on the basis of surveillance data.

A wild boar population infected with African Swine Fever (ASF) constitutes a constant threat to commercial pig farms and therefore to the economy of the affected country. Currently, ASF is still spreading in several countries and the implementation of intensive measures such as reducing wild boar population densities seems not to be able to stop the further spread of the disease. In addition, there are still substantial knowledge gaps regarding the epidemiology of the disease. To identify risk factors for a higher probability of a wild boar sample being virological or serological positive, comprehensive statistical analyses were performed based on Latvian surveillance data. Using a multivariable Bayesian regression model, the effects of implemented control measures on the proportion of hunted or found dead wild boar or on the estimated virus prevalence were evaluated. None of the control measures applied in Latvia showed a significant effect on the relevant target figure. Also, the estimated periodic prevalence of wild boar that had tested ASF positive by PCR appeared to remain unaffected over time. Therefore, there is an urgent need to reconsider the implemented control measures. The results of this study and the course of ASF in other affected countries, raise the question, whether an endemic situation of ASF in wild boar is reversible.

African swine fever virus introduction into the EU in 2014: Experience of Latvia.

African swine fever (ASF) virus was introduced in Latvia in June 2014. Thirty-two outbreaks in domestic pigs and 217 cases in wild boar were notified in 2014. Twenty-eight outbreaks (87.5%) were primary outbreaks. The contagiosity within pig herds was low. Failure to use simple biosecurity measures to reduce the chance of virus introduction, for example by inadvertent feeding of locally produced virus contaminated fodder were the main causes for the outbreaks in backyard holdings. The infection in wild boar survived locally in two different areas with a low prevalence and a slow spread. The persistence of the infection in wild boar within an area was most probably linked to wild boar scavenging the carcasses of infected wild boar.