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research priorities to fill knowledge gaps in wild boar management measures that could improve the control of african swine fever in wild boar populations.the european commission asked efsa to provide study designs for the investigation of four research domains (rds) according to major gaps in knowledge identified by efsa in a report published in 2019: (rd 1) african swine fever (asf) epidemiology in wild boar; (rd 2) asf transmission by vectors; (rd 3) african swine fever virus (asfv) survival in the environment, and (rd 4) the patterns of seasonality of asf in wild boar and domestic pigs in the eu. in this scientific opinion, the second rd on as ...202134354769
lateral flow assays for the detection of african swine fever virus antigen are not fit for field diagnosis of wild boar carcasses.african swine fever (asf) is one of the most important viral diseases of domestic pigs and wild boar. apart from endemic cycles in africa, asf is now continuously spreading in europe and asia. as asf leads to severe but unspecific clinical signs and high lethality, early pathogen detection is of utmost importance. recently, 'point-of-care' (poc) tests, especially immunochromatographic assays, have been intensively discussed for the use in remote areas but also in the context of on-farm epidemiol ...202134312995
rapid and sensitive rpa-cas12a-fluorescence assay for point-of-care detection of african swine fever virus.african swine fever (asf) is a serious contagious disease that causes fatal haemorrhagic fever in domestic and wild pigs, with high morbidity. it has caused devastating damage to the swine industry worldwide, necessitating the focus of attention on detection of the asf pathogen, the african swine fever virus (asfv). in order to overcome the disadvantages of conventional diagnostic methods (e.g. time-consuming, demanding and unintuitive), quick detection tools with higher sensitivity need to be e ...202134280234
a semi-automated system of luciferase immunoprecipitation assay for rapid and easy detection of african swine fever virus antibody.african swine fever (asf) is a highly contagious viral disease of domestic pigs and wild boars. for the disease surveillance and control, we developed a rapid and easy luciferase immunoprecipitation assay (mb-lips) to detect asf virus (asfv) antibody. the mb-lips is based on magnetic beads modified with protein a/g and the recombinant fusion protein of asfv p30 and luciferase, where p30 functioned as the recognition element and the luciferase as the signal component. incubation and washing could ...202134260273
evaluation of lesions and viral antigen distribution in domestic pigs inoculated intranasally with african swine fever virus ken05/tk1 (genotype x).the understanding of the pathogenic mechanisms and the clinicopathological forms caused by currently circulating african swine fever virus (asfv) isolates is incomplete. so far, most of the studies have been focused on isolates classified within genotypes i and ii, the only genotypes that have circulated outside of africa. however, less is known about the clinical presentations and lesions induced by isolates belonging to the other twenty-two genotypes. therefore, the early clinicopathological i ...202134207265
the role of interleukine-10 and interferon-γ as potential markers of the evolution of african swine fever virus infection in wild boar.african swine fever virus (asfv) is one of the most challenging pathogens to affect both domestic and wild pigs. the disease has now spread to europe and asia, causing great damage to the pig industry. although no commercial vaccine with which to control the disease is, as yet, available, some potential vaccine candidates have shown good results in terms of protection. however, little is known about the host immune mechanisms underlying that protection, especially in wild boar, which is the main ...202134203976
african swine fever virus as a difficult opponent in the fight for a vaccine-current data.prevention and control of african swine fever virus (asfv) in europe, asia, and africa seem to be extremely difficult in view of the ease with which it spreads, its high resistance to environmental conditions, and the many obstacles related to the introduction of effective specific immunoprophylaxis. biological properties of asfv indicate that the african swine fever (asf) pandemic will continue to develop and that only the implementation of an effective and safe vaccine will ensure a reduction ...202134201761
research priorities to fill knowledge gaps in the control of african swine fever: possible transmission of african swine fever virus by vectors.the european commission requested that efsa provide study designs for the investigation of four research domains according to major gaps in knowledge identified by efsa in a report published in 2019: (i) the patterns of seasonality of african swine fever (asf) in wild boar and domestic pigs in the eu; (ii) the epidemiology of asf in wild boar; (iii) survival of asf virus (asfv) in the environment and (iv) transmission of asfv by vectors. in this scientific opinion, the fourth research domain on ...202134188718
research objectives to fill knowledge gaps in african swine fever virus survival in the environment and carcasses, which could improve the control of african swine fever virus in wild boar populations.the european commission requested that efsa provide study designs for the investigation of four research domains according to major gaps in knowledge identified by efsa in a report published in 2019: i) the patterns of seasonality of african swine fever (asf) in wild boar and domestic pigs in the eu; ii) the epidemiology of asf in wild boar; iii) survival of asf virus (asfv) in the environment and iv) transmission of asfv by vectors. in this scientific opinion, the third research domain on asfv ...202134188717
transmission dynamics of african swine fever virus, south korea, 2019.african swine fever (asf) is a substantial concern for global food production and security. however, lack of epidemiologic data in affected areas has limited the knowledge of the main drivers of asf virus (asfv) transmission. to assess the role of vehicle movements and wild boar populations in spreading asfv to pig farms in south korea, we combined data generated by asf surveillance on pig farms and of wild boars with nationwide global positioning system-based tracking data for vehicles involved ...202134152953
the impact of african swine fever virus on smallholder village pig production: an outbreak investigation in lao pdr.african swine fever virus (asfv) causes a deadly disease of pigs which spread through southeast asia in 2019. we investigated one of the first outbreaks of asfv in lao people's democratic republic amongst smallholder villages of thapangtong district, savannakhet province. in this study, two asfv affected villages were compared to two unaffected villages. evidence of asfv-like clinical signs appeared in pig herds as early as may 2019, with median epidemic days on 1 and 18 june in the two villages ...202134146447
research progress on live attenuated vaccine against african swine fever virus.african swine fever (asf) is an acute, hemorrhagic and severe infectious disease caused by african swine fever virus (asfv) in domestic pigs and various wild boars, with a mortality rate up to 100%. asf was first discovered in 1921 in kenya. asfv has a large genome and complex immune escape mechanism creating difficulties in the production of vaccines. recently, remarkable advances have been made in vaccine development all over the world especially in live-attenuated vaccine. this article aims t ...202134089790
genome variability of classical swine fever virus during the 2018-2020 epidemic in japan.although rna viruses exhibit extensive sequence diversity, the mutation rate must be limited to ensure protein functions that maintain the viral life cycle. here, we compared the whole genome sequences of 150 isolates of classical swine fever virus (csfv), obtained from a single epidemic that occurred in japan during 2018-2020. after the detection of the first case, the disease spread among both farm pigs and wild boars and caused severe impact on the pig industry. to evaluate the diversificatio ...202134058522
comparative analysis of swine leukocyte antigen gene diversity in european farmed pigs.in europe, swine represent economically important farm animals and furthermore have become a preferred preclinical large animal model for biomedical studies, transplantation and regenerative medicine research. the need for typing of the swine leukocyte antigen (sla) is increasing with the expanded use of pigs as models for human diseases and organ-transplantation experiments and their use in infection studies and for design of veterinary vaccines. in this study, we characterised the sla class i ...202134028065
identification of a new cell-penetrating peptide derived from the african swine fever virus cd2v protein.the african swine fever virus (asfv) is a huge and complex dna virus that can lead to the acute death of pigs and cause huge losses to the global swine industry. the cd2v protein is a transmembrane protein encoded by the asfv's ep402r gene, which can effectively inhibit the bystander lymphocyte proliferation in response to mitogens and mediate the absorption of red blood cells to asfv-infected cells. the cd2v protein contains repetitive amino acid sequences ([kpcppp]3 labeled as raas), which is ...202134006158
comparative analysis of full genome sequences of african swine fever virus isolates taken from wild boars in russia in 2019.in this study, we report on the full genome phylogenetic analysis of four asfv isolates obtained from wild boars in russia. these samples originated from two eastern and two western regions of russia in 2019. phylogenetic analysis indicated that the isolates were assigned to genotype ii and grouped according to their geographical origins. the two eastern isolates shared 99.99% sequence identity with isolates from china, poland, belgium, and moldova, whereas the western isolates had 99.98% sequen ...202133925986
asfv-g-∆i177l as an effective oral nasal vaccine against the eurasia strain of africa swine fever.the african swine fever virus (asfv) is currently causing a pandemic affecting wild and domestic swine from western europe to asia. no commercial vaccines are available to prevent african swine fever (asf), resulting in overwhelming economic losses to the swine industry. we recently developed a recombinant vaccine candidate, asfvg-δi177l, by deleting the i177l gene from the genome of the highly virulent asfv strain georgia (asfv-g). asfv-g-δi177l has been proven safe and highly efficacious in ch ...202133925435
modelling the spatial distribution of asf-positive wild boar carcasses in south korea using 2019-2020 national surveillance data.in september 2019, african swine fever (asf) was reported in south korea for the first time. since then, more than 651 asf cases in wild boars and 14 farm outbreaks have been notified in the country. despite the efforts to eradicate asf among wild boar populations, the number of reported asf-positive wild boar carcasses have increased recently. the purpose of this study was to characterize the spatial distribution of asf-positive wild boar carcasses to identify the risk factors associated with t ...202133922261
african swine fever in cameroon: a review.african swine fever (asf) is a hemorrhagic contagious porcine disease caused by the african swine fever virus. the disease poses enormous problems to the pork industry with pig mortality ranging from 30% to 100%, depending on the virulence of the virus circulating. cameroon, situated in central africa is one of the countries in which the african swine fever virus (asfv) has been endemic since its first outbreak in 1982. the disease is a major problem to the pig industry causing huge economic los ...202133916101
phylogenetic and phylodynamic analysis of a classical swine fever virus outbreak in japan (2018-2020).after 26 years, another classical swine fever virus (csfv) outbreak in domestic pigs and wild boars occurred in japan 2018. herein, we investigated the entry and the spatial dynamics of the csfv outbreak in japan using the nearly complete genomes of strains isolated from both wild boars and domestic pigs during this epidemic. phylogenetic analysis showed that the most recent common ancestor (mrca) of the japanese lineage emerged 146 days (95% highest posterior density (hpd): 85-216 days) before ...202133890426
detection and quantification of african swine fever virus in ma-104 cells.detection of live african swine fever virus (asfv) has historically relied on the use of primary swine macrophages (psm). psm do not replicate and have to be isolated fresh from donor swine. we previously identified that a ma-104 cells (atcc #crl-2378.1), a commercially available cell line isolated from african green monkey ( cercopithecus aethiops ) kidney epithelial cells, supports the detection of asfv from field samples with a sensitivity comparable to that of primary swine macrophages. coll ...202133855107
rapid detection of african swine fever virus using cas12a-based portable paper diagnostics.african swine fever virus (asfv) is a dsdna virus responsible for a severe, highly contagious, and lethal disease affecting both domestic and wild pigs. asfv has brought enormous economic loss to a number of countries, and effective vaccine and therapy are still lacking. therefore, a rapid, sensitive, and field-deployable detection of asfv is important for disease surveillance and control. herein, we developed a cas12a-mediated portable paper assay to rapidly and precisely detect asfv. we identi ...202033824296
african swine fever virus genotype ii in mongolia, 2019.african swine fever (asf) is a severe haemorrhagic disease of domestic and wild pigs caused by the african swine fever virus (asfv). in recent years, asf has steadily spread towards new geographical areas, reaching europe and asia. on january 15th, 2019, mongolia reported its first asf outbreak to the world organization for animal health (oie), becoming, after china, the second country in the region affected by the disease. following an event of unusual mortality in domestic pigs in bulgan provi ...202133818903
the potential negative impacts of the classical swine fever virus on wild boar population in gifu prefecture, japan.there is a possibility that classical swine fever (csf) virus outbreak has negative impacts on wild boar. to adequately manage native wild boar populations, wildlife managers need to gather the field data on wild boar and implement population management practices. we aimed to report the relative abundance index of wild boar before and after this outbreak. our results showed that relative abundance index declined from 2017 (8.88 wild boars/100 trap days) to 2019 (2.03 wild boars/100 trap days), b ...202133775990
identification of african swine fever virus genomic dnas in wild boar habitats within outbreak regions in south korea.an african swine fever (asf) outbreak in wild boars was first reported on october 2, 2019, in south korea. since then, additional cases were reported in south korea's border areas. we here report the identification of asf virus (asfv) dnas from two out of eight environmental abiotic matter samples collected from areas where asf-positive wild boar carcasses were found. comparative genomic investigations suggested that the contaminating asfv dnas originated from the wild boar whose carcass had bee ...202133774943
subgrouping and analysis of relationships between classical swine fever virus identified during the 2018-2020 epidemic in japan by a novel approach using shared genomic variants.classical swine fever (csf) is a worldwide devastating disease of the pig industry caused by classical swine fever virus (csfv). in september 2018, an outbreak of csf occurred in japan where the disease had been eradicated and was officially designated a csf-free country since 2015. following the detection of the first 2018 case on a farm in gifu prefecture, the disease spread among both farm pigs and wild boars and still continues. epigenome analysis using whole-genome information is helpful in ...202133730417
asf exit strategy: providing cumulative evidence of the absence of african swine fever virus circulation in wild boar populations using standard surveillance measures.efsa assessed the role of seropositive wild boar in african swine fever (asf) persistence. surveillance data from estonia and latvia investigated with a generalised equation method demonstrated a significantly slower decline in seroprevalence in adult animals compared with subadults. the seroprevalence in adults, taking more than 24 months to approach zero after the last detection of asfv circulation, would be a poor indicator to demonstrate the absence of virus circulation. a narrative literatu ...202133717352
african swine fever virus mgf-110-9l-deficient mutant has attenuated virulence in pigs.african swine fever virus (asfv) is the etiological agent of african swine fever (asf), an often lethal disease in domestic and wild pigs. asf represents a major threat to the swine industry worldwide. currently, no commercial vaccine is available because of the complexity of asfv or biosecurity concerns. live attenuated viruses that are naturally isolated or genetically manipulated have demonstrated reliable protection against homologous asfv strain challenge. in the present study, a mutant asf ...202133689140
efficacy of oral vaccine against classical swine fever in wild boar and estimation of the disease dynamics in the quantitative approach.classical swine fever virus (csfv) in the wild boar population has been spreading in japan, alongside outbreaks on pigs, since classical swine fever (csf) reemerged in september 2018. the vaccination using oral bait vaccine was initially implemented in gifu prefecture in march 2019. in the present study, antibodies against csfv in wild boar were assessed in 1443 captured and dead wild boars in gifu prefecture. after the implementation of oral vaccination, the increase of the proportion of seropo ...202133672749
swift and reliable easy lab methods for the sensitive molecular detection of african swine fever virus.african swine fever (asf) is a contagious viral hemorrhagic disease of domestic pigs and wild boars. the disease is notifiable to the world organisation for animal health (oie) and is responsible for high mortality and serious economic losses. pcr and real-time pcr (qpcr) are the oie-recommended standard methods for the direct detection of african swine fever virus (asfv) dna. the aim of our work was the simplification and standardization of the molecular diagnostic workflow in the lab. for vali ...202133669073
an immortalized porcine macrophage cell line competent for the isolation of african swine fever virus.african swine fever virus (asfv) is the etiological agent of african swine fever (asf), a fatal hemorrhagic disease of domestic pigs and wild boar. the virus primarily infects macrophage and monocyte host cells, these do not grow in vitro. many attempts have been made to establish sustainable asfv-sensitive cell lines, but which supported only low viral replication levels of limited, mostly artificially attenuated strains of asfv. here, we examined the competence of a novel cell line of immortal ...202133637799
antibody prevalence to african swine fever virus, mycobacterium bovis, foot-and-mouth disease virus, rift valley fever virus, influenza a virus, and brucella and leptospira spp. in free-ranging warthog (phacochoerus africanus) populations in south africa.the warthog (phacochoerus africanus) can be used as a model for investigating disease transmission at the human, wildlife, and livestock interface. an omnivore and scavenger, a warthog moves freely between natural ecotypes, farmland, and human communities and is susceptible to diseases of zoonotic, agricultural, and conservation concern. a retrospective study using 100 individual serum samples collected from may 1999 to august 2016 was performed to determine antibody prevalence to seven pathogen ...202133635986
t-cell responses in domestic pigs and wild boar upon infection with the moderately virulent african swine fever virus strain 'estonia2014'.infection with african swine fever virus (asfv) causes a highly lethal haemorrhagic disease in domestic and eurasian wild pigs. thus, it is a major threat to pig populations worldwide and a cause of substantial economic losses. recently, less virulent asfv strains emerged naturally, which showed higher experimental virulence in wild boar than in domestic pigs. the reason for this difference in disease progression and outcome is unclear but likely involves different immunological responses. unfor ...202133630409
african swine fever spread across asia, 2018-2019.african swine fever virus (asfv) is a highly contagious pathogen that causes disease in pigs, commonly characterized by acute haemorrhagic fever. prior to august 2018, african swine fever (asf) had not been reported in asia, but has since spread throughout china, mongolia, korea, vietnam, laos, cambodia, myanmar, the philippines, hong kong, indonesia, timor-leste and papua new guinea. using data collated from reports of confirmed cases, we applied spatio-temporal analysis to describe asfv spread ...202133599077
self-amplifying pestivirus replicon rna encoding influenza virus nucleoprotein and hemagglutinin promote humoral and cellular immune responses in pigs.self-amplifying replicon rna (reprna) promotes expansion of mrna templates encoding genes of interest through their replicative nature, thus providing increased antigen payloads. reprna derived from the non-cytopathogenic classical swine fever virus (csfv) targets monocytes and dendritic cells (dcs), potentially promoting prolonged antigen expression in the dcs, contrasting with cytopathogenic reprna. we engineered pestivirus reprna constructs encoding influenza virus h5n1 (a/chicken/yamaguchi/7 ...202133584723
no evidence for african swine fever virus dna in haematophagous arthropods collected at wild boar baiting sites in estonia.african swine fever (asf) is a highly pathogenic viral disease affecting all suidae, with ornithodoros moubata complex soft ticks acting as the biological arthropod vectors of the causative agent, african swine fever virus (asfv). while asfv is also transmissible via direct contact, pig products and fomites, other arthropods may be involved in virus transmission and persistence. therefore, we checked various groups of blood-feeding arthropods collected during summer 2017 in wild boar habitats on ...202133527715
social structure defines spatial transmission of african swine fever in wild boar.the spatial spread of infectious disease is determined by spatial and social processes such as animal space use and family group structure. yet, the impacts of social processes on spatial spread remain poorly understood and estimates of spatial transmission kernels (stks) often exclude social structure. understanding the impacts of social structure on stks is important for obtaining robust inferences for policy decisions and optimizing response plans. we fit spatially explicit transmission model ...202133468025
molecular profile of african swine fever virus (asfv) circulating in vietnam during 2019-2020 outbreaks.african swine fever (asf) is a highly infectious disease of pigs caused by african swine fever virus (asfv). in order to identify potential genetic variations among asfv strains circulating in vietnam, 26 asfv isolates from organs and blood samples collected from domestic pigs from 23 different provinces of northern, central and southern vietnam during 2019-2020 asf outbreaks were genetically characterized. nucleotide sequences were determined for a portion of the b646l (p72) gene, the complete ...202133454861
unpicking the secrets of african swine fever viral replication sites.african swine fever virus (asfv) is a highly contagious pathogen which causes a lethal haemorrhagic fever in domestic pigs and wild boar. the large, double-stranded dna virus replicates in perinuclear cytoplasmic replication sites known as viral factories. these factories are complex, multi-dimensional structures. here we investigated the protein and membrane compartments of the factory using super-resolution and electron tomography. click it chemistry in combination with stimulated emission dep ...202133429879
african swine fever virus structural protein p17 inhibits cell proliferation through er stress-ros mediated cell cycle arrest.african swine fever virus (asfv) is a highly pathogenic large dna virus that causes african swine fever (asf) in domestic pigs and wild boars. the p17 protein, encoded by the d117l gene, is a major transmembrane protein of the capsid and the inner lipid envelope. the aim of this study was to investigate the effects of p17 on cell proliferation and the underlying mechanisms of action. the effects of p17 on cell proliferation, cell cycle, apoptosis, oxidative stress, and endoplasmic reticulum (er) ...202033374251
detection of african swine fever virus in free-ranging wild boar in southeast asia.african swine fever (asf) is a highly contagious and fatal viral disease affecting both domestic and wild suids. the virus was introduced to southeast asia in early 2019 and has since spread rapidly throughout the region. although significant efforts have been made to track and diagnose the disease in domestic pigs, very little is known about asf in free-ranging wild boar and their potential role in maintaining the disease within southeast asia. through a collaboration between government and non ...202033351995
distinct african swine fever virus shedding in wild boar infected with virulent and attenuated isolates.since the reappearance of african swine fever virus (asfv), the disease has spread in an unprecedented animal pandemic in eurasia. asf currently constitutes the greatest global problem for the swine industry. the wild boar (sus scrofa) in which the pathogen has established wild self-sustaining cycles, is a key reservoir for asfv, signifying that there is an urgent need to develop an effective vaccine against this virus. current scientific debate addresses whether live attenuated vaccines (lavs), ...202033339147
classical swine fever virus npro antagonises irf3 to prevent ifn-independent tlr3 and rig-i-mediated apoptosis.classical swine fever virus (csfv) is the causative agent of classical swine fever, a notifiable disease of economic importance that causes severe leukopenia, fever and haemorrhagic disease in domesticated pigs and wild boar across the globe. csfv has been shown to antagonise the induction of type i ifn, partly through a function of its n-terminal protease (npro) which binds irf3 and targets it for proteasomal degradation. additionally, npro has been shown to antagonise apoptosis triggered by th ...202033328306
assessment of risk factors of african swine fever in india: perspectives on future outbreaks and control strategies.african swine fever (asf) is one of the most important transboundary diseases of pigs. asf has been identified in india for the first time in domestic pigs from outbreaks reported in two of the northeastern states, arunachal pradesh and assam in 2020. a total of 11 asf outbreaks in different regions killed over 3700 pigs and devastated the economy of small-scale livestock owners of both the states. considering the first outbreak of asf in india, a generic risk assessment framework was determined ...202033322760
the contribution of insects to african swine fever virus dispersal: data from domestic pig farms in lithuania.outbreaks of african swine fever (asf) in domestic pig farms in lithuania typically begin in june and are detected through october, suggesting that insects might be involved in the transmission of the virus. entomological collecting was performed to obtain two data sets: from farms with asf outbreaks, and from farms without asf outbreaks but in an asf-infected area. target insects from the families muscidae, calliphoridae and tabanidae were analysed for the presence of asf virus (asfv) dna. musc ...202033314280
african swine fever: a permanent threat to indian pigs.india has 9 million pigs, of which 45% are in the north eastern (ne) states of india. viral diseases affecting pigs are a major concern of mortality causing huge loss to the pig farmers. one such disease is african swine fever (asf) that has already knocked the porous borders of ne states of india. asf is a highly contagious devastating disease of pigs and wild boars causing 100% mortality. the causative agent african swine fever virus (asfv) belongs to the genus asfivirus, family asfarviridae. ...202033281367
genetic analysis of african swine fever virus from the 2018 outbreak in south-eastern burundi.african swine fever (asf) is a contagious viral disease that causes high mortality, approaching 100%, in domestic pigs and wild boars. the disease has neither a cure nor a vaccine, and it is caused by an asf virus (asfv), the only member of the family asfarviridae, genus asfivirus, and the only known dna arbovirus. twenty-four genotypes of asfv have been described to date, and all of them have been described in africa. asf is endemic in burundi, and several outbreaks have been reported in the co ...202033251264
stability of african swine fever virus in soil and options to mitigate the potential transmission risk.understanding african swine fever virus (asfv) transmission is essential for strategies to minimize virus spread during an outbreak. asfv can survive for extended time periods in animal products, carcasses, and the environment. while the asfv genome was found in environments around infected farms, data on the virus survival in soil are scarce. we investigated different soil matrices spiked with asfv-positive blood from infected wild boar to see if asfv can remain infectious in the soil beneath i ...202033238521
achievements and challenges of classical swine fever eradication in brazil.classical swine fever virus (csfv) causes one of the most critical diseases in the porcine industry worldwide. in brazil, the first description of the infection was reported in 1888, and the national recognition of the first free zone (fz) occurred in 2001. brazil has been recently recognized (2015-2016) by the world organisation for animal health (oie) with an fz involving 15 states and the federal district, corresponding to 95% of the industrial production of pigs in the country, and a non-fre ...202033227889
the recombinant pseudorabies virus expressing african swine fever virus cd2v protein is safe and effective in mice.african swine fever (asf) leads to high mortality in domestic pigs and wild boar and is caused by the african swine fever virus (asfv). currently, no vaccine is commercially available for prevention, and the epidemic is still spreading. here, we constructed a recombinant pseudorabies virus (prv) (prv-δge/δgi/δtk-(cd2v)) that expresses the cd2v protein of asfv and evaluated its effectiveness and safety as a vaccine candidate in mice.202033198749
african swine fever virus mgf360-12l inhibits type i interferon production by blocking the interaction of importin α and nf-κb signaling pathway.african swine fever (asf) is an infectious transboundary disease of domestic pigs and wild boar and spreading throughout eurasia. there is no vaccine and treatment available. complex immune escape strategies of african swine fever virus (asfv) are crucial factors affecting immune prevention and vaccine development. mgf360 genes have been implicated in the modulation of the ifn-i response. the molecular mechanisms contributing to innate immunity are poorly understood. in this study, we demonstrat ...202033141406
whole-genome sequence of an african swine fever virus isolate from the czech republic.between june 2017 and april 2018, an outbreak of african swine fever (asf) affected wild boar in the southeast of the czech republic. here, we present the whole-genome sequence of the causative asf virus. it belongs to genotype ii and shows very high identity with other strains from eastern europe.202033122412
identification and isolation of two different subpopulations within african swine fever virus arm/07 stock.no efficient vaccines exist against african swine fever virus (asfv), which causes a serious disease in wild boars and domestic pigs that produces great industrial and ecological concerns worldwide. an extensive genetic characterization of the original asfv stocks used to produce live attenuated vaccine (lav) prototypes is needed for vaccine biosecurity and control. here, we sequenced for the first time the arm/07 stock which was obtained from an infected pig during the armenia outbreak in 2007, ...202033113838
fecal microbiota transplantation from warthog to pig confirms the influence of the gut microbiota on african swine fever susceptibility.african swine fever virus (asfv) is the causative agent of a devastating hemorrhagic disease (asf) that affects both domestic pigs and wild boars. conversely, asfv circulates in a subclinical manner in african wild pigs, including warthogs, the natural reservoir for asfv. together with genetic differences, other factors might be involved in the differential susceptibility to asf observed among eurasian suids (sus scrofa) and african warthogs (phacochoerus africanus). preliminary evidence obtaine ...202033077775
identification of african swine fever virus-like elements in the soft tick genome provides insights into the virus' evolution.african swine fever virus (asfv) is a most devastating pathogen affecting swine. in 2007, asfv was introduced into eastern europe where it continuously circulates and recently reached western europe and asia, leading to a socio-economic crisis of global proportion. in africa, where asfv was first described in 1921, it is transmitted between warthogs and soft ticks of the genus ornithodoros in a so-called sylvatic cycle. however, analyses into this virus' evolution are aggravated by the absence o ...202033032594
african swine fever in the lithuanian wild boar population in 2018: a snapshot.the first cases of african swine fever (asf) were detected in the lithuanian wild boar population in 2014. since then, the disease spread slowly through the whole country, affecting both, wild boar and domestic pigs. in the other baltic states, which both are also affected by asf since 2014, the recent course of asf prevalence suggests that the countries might be well under way of disease elimination. in contrast, in lithuania the epidemic seems to be still in full progress. in the present study ...202033028388
stability of african swine fever virus in carcasses of domestic pigs and wild boar experimentally infected with the asfv "estonia 2014" isolate.europe is currently experiencing a long-lasting african swine fever (asf) epidemic, both in domestic pigs and wild boar. there is great concern that carcasses of infected wild boar may act as long-term virus reservoirs in the environment. we evaluated the tenacity of asf virus (asfv) in tissues and body fluids from experimentally infected domestic pigs and wild boar, which were stored on different matrices and at different temperatures. samples were analysed at regular intervals for viral genome ...202033019736
rapid emergence of african swine fever virus variants with different numbers of a tandem repeat sequence in south korea.african swine fever virus variants with different numbers of a 10-bp tandem repeat were isolated in south korea soon after being identified in wild boar. the short emergence periods and sympatric distributions within a narrow geographical region suggest that the variants were sporadically generated in the pre-existing viral population.202033012101
persistent domestic circulation of african swine fever virus in tanzania, 2015-2017.african swine fever (asf) is a highly fatal viral hemorrhagic disease of domestic pigs that threatens livelihoods and food security. in africa, asf virus (asfv) circulates in sylvatic (transmission between warthogs and soft argasid ticks) and domestic (transmission between domestic pigs) cycles, with outbreaks resulting from asfv spill-over from sylvatic cycle. a number of outbreaks were reported in different parts of tanzania between 2015 and 2017. the present study investigated asfv transmissi ...202033004025
the spillover of african swine fever in western poland revealed its estimated origin on the basis of o174l, k145r, mgf 505-5r and igr i73r/i329l genomic sequences.the african swine fever epidemic occurred in poland at the beginning of 2014 and, up to date, the disease has been spreading mainly in the eastern part of the country. unexpectedly, in november 2019 an infected wild boar case was confirmed in lubuskie voivodship in western poland. during the following weeks, several dozen african swine fever virus (asfv)-positive animals were notified in the neighboring area, causing severe concern regarding further spread of the disease to the mostly pig-dense ...202032992547
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 teste ...202032967098
identification of novel testing matrices for african swine fever surveillance.african swine fever (asf) is a devastating viral disease of pigs and wild boar, and it threatens global food security. we aimed to identify suitable sample matrices for use in asf surveillance programs. six pigs inoculated with asfv were sampled at postmortem. blood, bone marrow, ear biopsies, and oral, nasal, and rectal swabs were taken from all pigs. all samples were analyzed using 3 real-time pcr (rtpcr) assays and a lamp assay. asfv was detected at > 107 genome copies/ml in blood; bone marro ...202032964810
genetically distinct pestiviruses pave the way to improved classical swine fever marker vaccine candidates based on the chimeric pestivirus concept.classical swine fever (csf) is one of the most important viral diseases of pigs. in many countries, the use of vaccines is restricted due to limitations of subunit vaccines with regard to efficacy and onset of protection as well as failure of live vaccines to differentiate infected from vaccinated animals (diva principle). chimeric pestiviruses based on csf virus (csfv) and the related bovine viral diarrhea virus (bvdv) have been licensed as live marker vaccines in europe and asia, but cross-rea ...202032962557
multifaceted immune responses to african swine fever virus: implications for vaccine development.african swine fever (asf) is a highly contagious, often fatal viral disease caused by african swine fever virus (asfv), leading to high fever, severe hemorrhages with high lethality in domestic pigs and wild boar. in 2007, asf was reintroduced into europe. since then, asf has spread to many european and asian countries and now becomes a major concern to the swine industry worldwide. there have been various vaccine attempts, but no commercial asf vaccines are available so far. a key hurdle in dev ...202032932135
porcine immunoglobulin fc fused p30/p54 protein of african swine fever virus displaying on surface of s. cerevisiae elicit strong antibody production in swine.african swine fever virus (asfv) infects domestic pigs and european wild boars with strong, hemorrhagic and high mortality. the primary cellular targets of asfv is the porcine macrophages. up to now, no commercial vaccine or effective treatment available to control the disease. in this study, three recombinant saccharomyces cerevisiae (s. cerevisiae) strains expressing fused asfv proteins-porcine ig heavy chains were constructed and the immunogenicity of the s. cerevisiae-vectored cocktail asfv ...202032915442
isolation and propagation of classical swine fever virus in porcine wharton's jelly mesenchymal stem cells.classical swine fever (csf) is an extremely infectious and deadly disease of pigs and wild boars caused by the csf virus (csfv) which is a member of the pestivirus genus and the family flaviviridae. this study was designed to detect the permissibility and replication of csfv in mesenchymal stem cells (mscs) monolayer derived from porcine wharton's jelly. porcine wharton's jelly mscs (pwj-mscs) were ex vivo expanded and propagated for more than 81 generations and third passage pwj-mscs were chara ...202032885744
a wide-ranging antiviral response in wild boar cells is triggered by non-coding synthetic rnas from the foot-and-mouth disease virus genome.foot-and-mouth disease virus (fmdv) is the causative agent of a highly contagious viral disease that affects multiple cloven-hooved hosts including important livestock (pigs, cattle, sheep and goats) as well as several wild animal species. crossover of fmdv between domestic and wildlife populations may prolong virus circulation during outbreaks. the wild boar (sus scrofa) is considered a reservoir of various pathogens that can infect other wildlife, domestic animals, and humans. as wild boar and ...202032851049
clinical course and gross pathological findings in wild boar infected with a highly virulent strain of african swine fever virus genotype ii.african swine fever (asf) is a notifiable disease that in recent years has spread remarkably in europe and asia. eurasian wild boar (sus scrofa) plays a key role in the maintenance and spread of the pathogen. here we examined gross pathology of infection in wild boar with a highly virulent, hemadsorbing genotype ii asf virus (asfv) strain. to this end, six wild boars were intramuscularly inoculated with the 10 had50 arm07 asfv strain, and 11 wild boars were allowed to come into direct contact wi ...202032842614
evaluation of un-methylated dna enrichment in sequencing of african swine fever virus complete genome.african swine fever is a febrile hemorrhagic fever disease that is caused by the african swine fever virus (asfv) and is lethal for domestic pigs and wild boar. asfv also infects soft ticks of the genus ornithodoros, some species of which can act as a vector for asfv. whole genome sequencing of asfv is a challenge because, due to the size difference of the host genome versus the viral genome, the higher proportion of host versus virus dna fragments renders the virus sequencing poorly efficient. ...202032828806
comparative pathology of domestic pigs and wild boar infected with the moderately virulent african swine fever virus strain "estonia 2014".endemically infected european wild boar are considered a major reservoir of african swine fever virus in europe. while high lethality was observed in the majority of field cases, strains of moderate virulence occurred in the baltic states. one of these, "estonia 2014", led to a higher number of clinically healthy, antibody-positive animals in the hunting bag of north-eastern estonia. experimental characterization showed high virulence in wild boar but moderate virulence in domestic pigs. putativ ...202032824331
clinical validation of two recombinase-based isothermal amplification assays (rpa/raa) for the rapid detection of african swine fever virus.african swine fever (asf), caused by african swine fever virus (asfv), is a devastating infectious disease of domestic pigs and wild boars, and has tremendous negative socioeconomic impact on the swine industry and food security worldwide. it is characterized as a notifiable disease by world organisation for animal health (oie). no effective vaccine or treatment against asf has so far been available. early detection and rapid diagnosis are of potential significance to control the spread of asf. ...202032793160
expression of recombinant classical swine fever virus e2 glycoprotein by endogenous txnip promoter in stable transgenic cho cells.as the main immunogen that could stimulate neutralized antibody in pigs, recombinant e2 protein of csfv was expressed in cho-dhfr-cells driven by endogenous txnip promoter from chinese hamster. different fragments of txnip promoter were amplified by pcr from isolated genomic dna of cho cells and cloned into different expression vectors. compared with cmv promoter, cho-ptxnip-4-re2 (f12) cell clone with the highest yield of re2 protein was established by random insertion of the expression cassett ...202032774204
efficient inactivation of african swine fever virus by ozonized water.african swine fever virus (asfv) is the causative agent of african swine fever (asf), which is a devastating disease of domestic pigs and wild boar, causing significant economic losses to the pig industry worldwide. to evaluate the ability of ozonized water as a disinfectant to inactivate asfv, ozonized water of different concentrations was tested, and the viral reduction was determined by infectivity assay on porcine primary alveolar macrophages. the results showed that 2 log10 (99 %) reduction ...202032768237
genome sequences of three african swine fever viruses of genotypes iv and xx from zaire and south africa, isolated from a domestic pig (sus scrofa domesticus), a warthog (phacochoerus africanus), and a european wild boar (sus scrofa).we report here the genome sequences of three african swine fever virus isolates obtained from a domestic pig (zaire [zaire]), a warthog (rsa/w1/1999 [south africa]), and a european wild boar (rsa/2/2004 [south africa]) belonging to genotypes iv, xx, and xx, respectively. this report increases the number of genotype xx, wild boar, and warthog reference sequences available.202032763924
an expert opinion assessment of blood-feeding arthropods based on their capacity to transmit african swine fever virus in metropolitan france.to deal with the limited literature data on the vectorial capacity of blood-feeding arthropods (bfas) and their role in the transmission of african swine fever virus (asfv) in metropolitan france, a dedicated working group of the french agency for food, environmental and occupational health & safety performed an expert knowledge elicitation. in total, 15 different bfas were selected as potential vectors by the ad hoc working group involved. ten criteria were considered to define the vectorial ca ...202032750188
toward better control of classical swine fever in wild boars: susceptibility of boar-pig hybrids to a recent japanese isolate and effectiveness of a bait vaccine.we analyzed the pathogenicity of a recent japanese classical swine fever virus (csfv) to wild boars via an experimental infection using boar-pig hybrids as an alternative to wild boars. we also investigated the effectiveness of a bait vaccine against the csfv. naïve boar-pig hybrids and pigs showed clinical signs such as fever, leucopenia, anorexia and conjunctivitis following the experimental infection. in contrast, the boar-pig hybrids administered the bait vaccine did not show any clinical si ...202032736657
identification of a continuously stable and commercially available cell line for the identification of infectious african swine fever virus in clinical samples.african swine fever virus (asfv) is causing outbreaks both in domestic pigs and wild boar in europe and asia. in 2018, the largest pig producing country, china, reported its first outbreak of african swine fever (asf). since then, the disease has quickly spread to all provinces in china and to other countries in southeast asia, and most recently to india. outbreaks of the disease occur in europe as far west as poland, and one isolated outbreak has been reported in belgium. the current outbreak s ...202032731642
health profile of free-range wild boar (sus scrofa) subpopulations hunted in santa catarina state, brazil.wild boars represent a potential dissemination source of important pathogens to public and animal health, since they can host several pathogens. this study aimed to evaluate the health profile of wild boars hunted for population control in the state of santa catarina (sc), south of brazil. for this study, tissue and blood samples were collected from 61 wild boars hunted from october 2017 to november 2018. all 61 serum samples were screened for antibodies by different assays, presenting the follo ...202032725949
molecular characterization of african swine fever virus isolates in estonia in 2014-2019.after the extensive spread of the african swine fever virus (asfv) genotype ii in eastern europe, the first case of african swine fever (asf) in estonia was diagnosed in september 2014. by the end of 2019, 3971 asfv-positive wild boars were found, and 27 domestic pig outbreaks were reported. a selection of asfv isolates from wild boar and domestic pigs (during the period of september 2014-2019) was molecularly characterized using standardized genotyping procedures. one of the proven markers to c ...202032709020
putative role of arthropod vectors in african swine fever virus transmission in relation to their bio-ecological properties.african swine fever (asf) is one of the most important diseases in suidae due to its significant health and socioeconomic consequences and represents a major threat to the european pig industry, especially in the absence of any available treatment or vaccine. in fact, with its high mortality rate and the subsequent trade restrictions imposed on affected countries, asf can dramatically disrupt the pig industry in afflicted countries. in september 2018, asf was unexpectedly identified in wild boar ...202032698448
detection of african swine fever virus genotype xv in a sylvatic cycle in saadani national park, tanzania.african swine fever (asf) is a severe hemorrhagic disease of domestic pigs caused by asf virus (asfv). asfv is transmitted by soft ticks (ornithodoros moubata complex group) and by direct transmission. in africa, asf is maintained in transmission cycles of asymptomatic infection involving wild suids, mainly warthogs (phacochoerus africanus). asf outbreaks have been reported in many parts of tanzania; however, active surveillance has been limited to pig farms in a few geographical locations. ther ...202032696552
research gap analysis on african swine fever.the most significant knowledge gaps in the prevention and control of african swine fever (asf) were identified by the eu veterinary services and other stakeholders involved in pig production and wild boar management through an online survey. the respondents were asked to identify the major research needs in order to improve short-term asf risk management. four major gaps were identified: 'wild boar', 'african swine fever virus (asfv) survival and transmission', 'biosecurity' and 'surveillance'. ...201932626417
epidemiological analyses on african swine fever in the baltic countries and poland.african swine fever virus (asfv) has been notified in the baltic countries and the eastern part of poland from the beginning of 2014 up to now. in collaboration with the asf-affected member states (ms), efsa is updating the epidemiological analysis of asf in the european union which was carried out in 2015. for this purpose, the latest epidemiological and laboratory data were analysed in order to identify the spatial-temporal pattern of the epidemic and a risk factors facilitating its spread. cu ...201732625438
epidemiological analyses of african swine fever in the baltic states and poland: (update september 2016-september 2017).efsa assisted four countries in the analysis of epidemiological data on african swine fever (asf), collected until september 2017. the temporal analysis demonstrated that the average proportions of pcr and antibody-elisa positive samples from the hunted wild boar remained below 3.9 and 6.6, respectively. a peak in the asf incidence was observed 6 months after the first observed case, followed by a significant reduction of the number of cases and low levels of african swine fever virus (asfv) cir ...201732625356
one year of african swine fever outbreak in china.african swine fever (asf) is a major threat to domestic pigs and wild boars. since 2018, asf outbreaks have been ongoing in china. as of august 3, 2019, a total of 151 asf clusters of outbreaks reported in china have caused severe economic losses for the industry, the pig farmers and pork producers, due to the lack of an efficacious vaccine. the present study aims to analyze the epidemiologic characteristics of asf outbreak that occurred in several regions across china during the period august 2 ...202032598922
attempts at the development of a recombinant african swine fever virus strain with abrogated ep402r, 9gl, and a238l gene structure using the crispr/cas9 system.african swine fever (asf) is a pressing economic problem in a number of eastern european countries. it has also depleted the chinese sow population by 50%. managing the disease relies on culling infected pigs or hunting wild boars as sanitary zone creation. the constraints on the development of an efficient vaccine are mainly the virus' mechanisms of host immune response evasion. the study aimed to adapt a field asfv strain to established cell lines and to construct recombinant african swine fev ...202032587905
impaired t-cell responses in domestic pigs and wild boar upon infection with a highly virulent african swine fever virus strain.since african swine fever (asf) first appeared in the caucasus region in 2007, it has spread rapidly and is now present in numerous european and asian countries. in europe, mainly wild boar populations are affected and pose a risk for domestic pigs. in asia, domestic pigs are almost exclusively affected. an effective and safe vaccine is not available, and correlates of protection are far from being understood. therefore, research on immune responses, immune dysfunction and pathogenesis is mandat ...202032530090
a recombinase polymerase amplification combined with lateral flow dipstick for rapid and specific detection of african swine fever virus.african swine fever (asf) is an acute, hemorrhagic, highly contagious disease caused by african swine fever virus (asfv) infection of domestic pigs and wild boars, showing mortality rates up to 100 %. there are no effective vaccines or antiviral drugs available for asfv. therefore, disease control is mainly based on animal slaughtering and the enforcement of strict sanitary measures. in order to establish a rapid, sensitive and simple method for on-site detection of asfv, a recombinase polymeras ...202032492462
substitution of warthog nf-κb motifs into rela of domestic pigs is not sufficient to confer resilience to african swine fever virus.african swine fever virus (asfv) causes a lethal, haemorrhagic disease in domestic swine that threatens pig production across the globe. unlike domestic pigs, warthogs, which are wildlife hosts of the virus, do not succumb to the lethal effects of infection. there are three amino acid differences between the sequence of the warthog and domestic pig rela protein; a subunit of the nf-κb transcription factor that plays a key role in regulating the immune response to infections. domestic pigs with a ...202032488046
african swine fever virus: an emerging dna arbovirus.african swine fever virus (asfv) is the sole member of the family asfarviridae, and the only known dna arbovirus. since its identification in kenya in 1921, asfv has remained endemic in africa, maintained in a sylvatic cycle between ornithodoros soft ticks and warthogs (phacochoerus africanus) which do not develop clinical disease with asfv infection. however, asfv causes a devastating and economically significant disease of domestic (sus scrofa domesticus) and feral (sus scrofa ferus) swine. th ...202032478103
stability of african swine fever virus on heat-treated field crops.african swine fever (asf) is an infectious disease of pigs and represents a massive threat to animal health and the pig industry worldwide. the asf virus (asfv) is efficiently transmitted via blood and meat from infected animals and can be highly stable in the environment. there is therefore great concern about the potential role of contaminated raw materials used for feed or bedding in the spread of asfv. especially crops and derived products originating from areas with asf in wild boar and thu ...202032460443
detection of african swine fever virus in cell culture and wild boar tissues using a commercially available monoclonal antibody.following the introduction of african swine fever virus (asfv) into europe in 2007, asfv infection has spread continuously over the past years and it became a high level disease threat in europe and also asia. examination of suspect clinical cases for asf with rapid and sensitive laboratory methods can substantially contribute to the detection and characterization of new outbreaks. in this study two sensitive tests were developed for the detection of the p72 major capsid protein of asfv both in ...202032450150
monitoring relative abundance index and age ratios of wild boar (sus scrofa) in small scale population in gifu prefecture, japan during classical swine fever outbreak.although the first cases of classical swine fever were reported in 2018, no studies have explored this impact on wild boar populations in japan. comparing the relative abundance indices and age ratios in the wild boar population before and after the outbreak, we investigated the impact of classical swine fever virus on wild boar population dynamics in august 2017-december 2019. relative abundance indices declined from 2017 to 2019 drastically, while there were no significant differences in age r ...202032435014
african swine fever in a commercial pig farm: outbreak investigation and an approach for identifying the source of infection.african swine fever (asf) is a contagious disease of domestic and wild pigs caused by the african swine fever virus (asfv). the disease has spread globally in recent years with serious economic consequences to pork production. this report describes an asf outbreak that occurred in a large-scale chinese commercial pig farm. the outbreak started in 2018 and presents the spatial and temporal spread of infection in an intensive pig farm. pig houses adjacent to exit rooms had a higher risk of infecti ...202032419367
evaluation of seven commercial african swine fever virus detection kits and three taq polymerases on 300 well-characterized field samples.african swine fever virus (asfv) is a complex double stranded dna virus, responsible for a highly infectious and fatal disease in pigs and boars and for important deterioration of animal welfare. over the last decade, the disease spread to several european and asian countries causing unprecedented dramatic economic losses in pig industry. in the absence of a vaccine, affected countries rely on trustful diagnostic tests and adapted testing policies to set up control programs to fight against the ...202032360149
development of a triplex real-time pcr assay for detection and differentiation of gene-deleted and wild-type african swine fever virus.african swine fever (asf) is an infectious disease of domestic and wild pigs, caused by asf virus (asfv). in this study, a triplex real-time pcr assay was developed to detect and differentiate the gene-deleted and wild-type asfv strains. three pairs of primers and probes were designed to target the conserved region of b646l gene (p72), mgf_360-14l gene (located in the middle of mgf360-505r gene) and cd2v gene, respectively. gene-deleted (with mgf360-505r and / or cd2v genes deletion) and wild-ty ...202032333943
risk assessment of african swine fever virus exposure to sus scrofa in japan via pork products brought in air passengers' luggage.in recent years, african swine fever (asf) has become prevalent in many areas, including asia. the repeated detection of the asf virus (asfv) genome in pork products brought in air passenger's luggage (ppap) was also reported from japanese airports. in the present study, the risk of asfv exposure to susceptible hosts in japan via three different pathways was assessed. two quantitative stochastic risk assessment models were built to estimate the annual probability of asfv exposure to domestic pig ...202032326040
the epidemiological status of african swine fever in domestic swine herds in the tavush province region, armenia.keen the factors associated with the spread and persistence of african swine fever (asf) in the caucasus region remain to be fully identified. it is assumed that large naive populations of domestic free-ranging and wild pigs are critical to disease transmission and maintenance. nonetheless, 11 years since its epidemic introduction into the region in 2007, asf virus (asfv) is still circulating, suggesting that an endemic cycle has been established based on contact between free-ranging domestic pi ...201932286569
rapid detection of african swine fever virus using cas12a-based portable paper diagnostics.african swine fever virus (asfv) is a dsdna virus responsible for a severe, highly contagious, and lethal disease affecting both domestic and wild pigs. asfv has brought enormous economic loss to a number of countries, and effective vaccine and therapy are still lacking. therefore, a rapid, sensitive, and field-deployable detection of asfv is important for disease surveillance and control. herein, we developed a cas12a-mediated portable paper assay to rapidly and precisely detect asfv. we identi ...202032284877
african swine fever virus - the possible role of flies and other insects in virus transmission.african swine fever (asf) is an acute viral haemorrhagic disease of pigs and wild boars. it presents a serious threat to pig production worldwide, and since 2007, asf outbreaks have been recorded in the caucasus, eastern europe, and the baltic states. in 2014, the disease was detected in poland. asf is on the list of notifiable diseases of the world organisation for animal health (oie). due to the lack of an available vaccine and treatment, the countermeasures against the disease consist in earl ...202032258793
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