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.

Evidence-Based African Swine Fever Policies: Do We Address Virus and Host Adequately?

African swine fever (ASF) is one of the most threatening diseases for the pig farming sector worldwide. Prevention, control and eradication remain a challenge, especially in the absence of an effective vaccine or cure and despite the relatively low contagiousness of this pathogen in contrast to Classical Swine Fever or Foot and Mouth disease, for example. Usually lethal in pigs and wild boar, this viral transboundary animal disease has the potential to significantly disrupt global trade and threaten food security. This paper outlines the importance of a disease-specific legal framework, based on the latest scientific evidence in order to improve ASF control. It compares the legal basis for ASF control in a number of pig-producing regions globally, considering diverse production systems, taking into account current scientific evidence in relation to ASF spread and control. We argue that blanket policies that do not take into account disease-relevant characteristics of a biological agent, nor the specifics under which the host species are kept, can hamper disease control efforts and may prove disproportionate.

Mathematical Approach to Estimating the Main Epidemiological Parameters of African Swine Fever in Wild Boar.

African swine fever (ASF) severely threatens the swine industry worldwide, given its spread and the absence of an available licensed vaccine, and has caused severe economic losses. Its persistence in wild boar (WB), longer than in domestic pig farms, and the knowledge gaps in ASF epidemiology hinder ASF virus (ASFV) eradication. Even in areas where disease is effectively controlled and ASFV is no longer detected, declaring eradication is difficult as seropositive WBs may still be detected. The aim of this work was to estimate the main ASF epidemiological parameters specific for the north of Sardinia, Italy. The estimated basic (R0) and effective (Re) reproduction numbers demonstrate that the ASF epidemic is declining and under control with an R0 of 1.139 (95% confidence interval (CI) = 1.123-1.153) and Re of 0.802 (95% CI = 0.612-0.992). In the last phases of an epidemic, these estimates are crucial tools for identifying the intensity of interventions required to definitively eradicate the disease. This approach is useful to understand if and when the detection of residual seropositive WB is no longer associated with any further ASFV circulation.

African Swine Fever Circulation among Free-Ranging Pigs in Sardinia: Data from the Eradication Program.

African swine fever virus (ASFV), the cause of a devastating disease affecting domestic and wild pigs, has been present in Sardinia since 1978. In the framework of the regional ASF eradication plan, 4484 illegal pigs were culled between December 2017 and February 2020. The highest disease prevalence was observed in the municipality with the highest free-ranging pig density, and culling actions drastically reduced ASFV circulation among these animals. ASFV-antibody were detected in 36.7% of tested animals, which were apparently healthy, thus, the circulation of low-virulence ASFV isolates was hypothesized. ASFV genome was detected in 53 out of 2726 tested animals, and virus isolation was achieved in two distinct culling actions. Two ASFV haemadsorbing strains were isolated from antibody-positive apparently healthy pigs: 55234/18 and 103917/18. Typing analysis revealed that both isolates belong to p72 genotype I, B602L subgroup X; phylogenetic analysis based on whole genome sequencing data showed that they were closely related to Sardinian ASFV strains collected since 2010, especially 22653/Ca/2014. Our data suggested the absence of immune-escaped ASFV variants circulating among free-ranging pigs, indicating that other elements contributed to virus circulation among these animals. Understanding factors behind disease persistence in endemic settings might contribute to developing effective countermeasures against this disease.

Evaluation of the cost-effectiveness of ASF detection with or without the use of on-field tests in different scenarios, in Sardinia.

African swine fever (ASF) is a highly contagious disease of domestic pigs and wild boars (WBs). Without a vaccine, early antibody and antigen detection and rapid diagnosis are crucial for the effective prevention of the disease and the employment of control measures. In Sardinia, where 3 different suid populations coexisted closely for a long time, the disease persists since 1978. The recent ASF eradication plan involves more stringent measures to combat free-ranging pigs and any kind of illegality in the pig industry. However, critical issues such as the low level of hunter cooperation with veterinary services and the time required for ASF detection in the WBs killed during the hunting season still remain. Considering the need to deliver true ASF negative carcasses as early as possible, this study focuses on the evaluation and validation of a duplex pen-side test that simultaneously detects antibodies and antigens specific to ASF virus, to improve molecular diagnosis under field conditions. The main goal was to establish the specificity of the two pen-side tests performed simultaneously and to determine their ability to detect the true ASF negative carcasses among the hunted WBs. Blood and organ samples of the WBs hunted during the 2018/2019 hunting seasons were obtained. A total of 160 animals were tested using the pen-side kit test; samples were collected for virological and serological analyses. A specificity of 98% was observed considering the official laboratory tests as gold standards. The new diagnostic techniques could facilitate faster and cost-effective control of the disease.

Correction: Socio-economic factors as indicators for various animal diseases in Sardinia.

[This corrects the article DOI: 10.1371/journal.pone.0217367.].

Socio-economic factors as indicators for various animal diseases in Sardinia.

The need to consider the role of social factors in the efficacy of farm management and, consequently, in the onset and persistence of diseases typical to animal farms is increasingly being realized increasingly worldwide. Many risk analysis studies have been conducted to assess the role of various factors in the development of animal diseases; however, very few have accounted for the role of social factors. The aim of this work was to bridge this gap, with the main hypothesis that different socio-economic factors could be valid indicators for the occurrence of different animal diseases. A socio-economic analysis was performed using demographic characteristics of the farmers and data from 44 social indicators released by the Italian Statistician National Institute of Statistics (ISTAT) database. African swine fever (ASF) in wild boars (WB) and domestic pigs and other endemic animal diseases and zoonoses in Sardinia were considered, such as cistic echinococcosis (CE), contagious agalactia (CA), trichinellosis, West Nile disease (WND), and bluetongue (BT). Seven different negative binomial regression models were fitted using the number of cases between 2011-2017. Three indicators-cultural demand, employment rate, and legality-showed a statistically significant association with risk for all the diseases considered, but with varying effects. Some indicators, such as the age and sex of the farmer, material deprivation index, number of farms and animals, micro-criminality index, and rate of reported thefts were common to ASF, CA, trichinellosis, and CE cases. Others such as the forest surface and the energy produced from renewable sources were common to BT, WND, and ASF in WB. Tourism in seasons other than summer was a valid predictor of ASF and trichinellosis, while out-of-region hospital use had a statistically significant role in CE risk identification. These results may help understand the social context in which these diseases may occur and thus guide the design and implementation of additional risk management measures that go beyond well-known veterinary measures.

Surveillance and control of African Swine Fever in free-ranging pigs in Sardinia.

African swine fever (ASF) is a notifiable infectious disease, caused by the ASF virus (ASFV), which is a DNA virus belonging to the family Asfarviridae, genus Asfivirus. This disease has gained importance in the last decade after its spread in several countries in Eastern and Central Europe, and more recently, in China. Despite the efforts made to eradicate it, ASF is still present on the Mediterranean island of Sardinia (Italy) and has been since 1978. ASF risk factors on the island have been analysed in previous studies; the role of free-ranging pigs in virus persistence has been suggested, but has not been fully elucidated. The most recent eradication plan provides more stringent measures to combat free-ranging pigs and any kind of illegality in the pig sector. From December 2017 to June 2018, a total of 29 depopulation actions were performed in 13 municipalities in central Sardinia, during which 2,281 free-ranging pigs were culled and more than 50% of them were tested for ASFV and antibody presence (1,218 and 1,416, respectively). A total of 651 pigs were seropositive, with a mean seroprevalence of 53.4% (CI 95% = 50.6-56.3), and 38 were ASFV positive (virus prevalence = 2.6%; CI 95% = 2.1-3.0). To the best of our knowledge, the present study is the first to provide a complete evaluation of this millennial system of pig farming and ASFV prevalence in free-ranging pigs. Furthermore, it has emphasised the necessity of combining the maintenance of an epidemiological surveillance program with continuous education of farmers and other people involved in pig husbandry, based on cultural and economic aspects.

Evaluation of biological and socio-economic factors related to persistence of African swine fever in Sardinia.

Despite the implementation of several eradication programmes, African swine fever (ASF), a viral disease in pigs caused by a DNA virus (ASFV), has been present in Sardinia (Italy) since 1978. Several studies have been carried out on the epidemiology of ASF in Sardinia, aimed at attaining a better understanding of the role of the risk factors related to ASFV persistence, but those studies did not address the social aspects involved. This work sought to bridge this gap, identifying the main social risk factors associated with ASF persistence. With this aim, this study takes into account not only the known "biological" risk factors identified in previous studies, but also the direct correlation between ASF persistence and well-known socio-economic aspects. The demographic characteristics, the Material Deprivation Index (IDM) and the non-compliance with the rules on ASF controls, including the traditional method of keeping free-range pigs has been evaluated. To assess the weight of each risk factor, data about pig farms, wild boar and social factors in Sardinia, were analysed using the Negative Binomial Regression Model. The main outcome was the number of domestic pig outbreaks occurring in Sardinian during 2011-2016. The effect in terms of the odds ratio (OR) was calculated to each factor included. The biological risk factors identified covered the number of animals (OR = 3.33, p < .0001, by 100 animals) and farms (OR = 1.07, p = .006, by 10 farms), the animal movements (OR = 1.64, p = .001, by 10 movements), the presence of illegal pigs (OR = 6.87, p < .0001) and the ASFV prevalence in wild boars (OR = 1.30, p = .001). Among the socio-economic factors, the compliance with control measures (OR = 0.90, p < .0001), the human population increasing by 1000 people (OR = 0.89, p < .0001), the growing age of the farmers (OR = 0.66, p = .025, by 5 years) and non-relationships with other farms (OR = 0.85, p < .001), decreased the ASF risk. The deprived condition (i.e. cultural and material deprivation, lack of resources and overcrowding index) increases the risk of about four times, as the low educational level (OR = 3.97, p = .002). Having highlighted the important role of social conditions, this risk definition allows understanding the Sardinian situation and may be useful to decision-makers to draft specific control strategies against this disease in the island, which should take into account local risk factors.

Approaches and Perspectives for Development of African Swine Fever Virus Vaccines.

African swine fever (ASF) is a complex disease of swine, caused by a large DNA virus belonging to the family Asfarviridae. The disease shows variable clinical signs, with high case fatality rates, up to 100%, in the acute forms. ASF is currently present in Africa and Europe where it circulates in different scenarios causing a high socio-economic impact. In most affected regions, control has not been effective in part due to lack of a vaccine. The availability of an effective and safe ASFV vaccines would support and enforce control-eradication strategies. Therefore, work leading to the rational development of protective ASF vaccines is a high priority. Several factors have hindered vaccine development, including the complexity of the ASF virus particle and the large number of proteins encoded by its genome. Many of these virus proteins inhibit the host's immune system thus facilitating virus replication and persistence. We review previous work aimed at understanding ASFV-host interactions, including mechanisms of protective immunity, and approaches for vaccine development. These include live attenuated vaccines, and "subunit" vaccines, based on DNA, proteins, or virus vectors. In the shorter to medium term, live attenuated vaccines are the most promising and best positioned candidates. Gaps and future research directions are evaluated.

Characterization of the interaction of African swine fever virus with monocytes and derived macrophage subsets.

African swine fever (ASF) is a devastating disease for which there is no vaccine available. The ASF virus (ASFV) primarily infects cells of the myeloid lineage and this tropism is thought to be crucial for disease pathogenesis. A detailed in vitro characterization of the interactions of a virulent Sardinian isolate (22653/14) and a tissue culture adapted avirulent strain (BA71V) of ASFV with porcine monocytes, un-activated (moMΦ), classically (moM1) and alternatively (moM2) activated monocyte-derived macrophages was conducted in an attempt to better understand this relationship. Using a multiplicity-of-infection (MOI) of 1, both viruses were able to infect monocytes and macrophage subsets, but BA71V presented a reduced ability to infect moM1 compared to 22653/14, with higher expression of early compared to late proteins. Using an MOI of 0.01, only 22653/14 was able to replicate in all the macrophage subsets, with initially lowest in moM1 and moM2. No differences were observed in the expression of CD163 between ASFV infected and uninfected bystander cells. ASFV down-regulated CD16 expression but did not modulate MHC class II levels in monocytes and macrophage subsets. BA71V-infected but not 22653/14-infected moMΦ and moM2 presented with a reduced expression of MHC class I compared to the mock-infected controls. Higher levels of IL-18, IL1-ß and IL-1α were released from moM1 after infection with BA71V compared to 22653/14 or mock-infected control. These results revealed differences between these ASFV strains, suggesting that virulent isolates have evolved mechanisms to counteract activated macrophages responses, promoting their survival, dissemination in the host and so ASF pathogenesis.