A bread‐based lyophilized C‐strain CSF virus vaccine as an oral vaccine in pigs

A live attenuated cereal‐based classical swine fever (CSF) vaccine for oral application, a form of vaccine that farmers can use themselves, demonstrated the improvement of CSF control in backyard production systems in endemic areas. Due to the dependency on very low storage temperature (−20°C) of the cereal‐based oral CSF vaccine, a lyophilized cereal‐based oral vaccine has been developed and tested. Although some studies showed total protection against a virulent virus strain, the production procedure is still considered complex. In this study, the lyophilized oral CSF virus, which is easy to produce and could be kept in the form of a bread‐based vaccine at 4°C for an extended time, was tested for the ability to induce a humoral immune response in pigs. Among the materials tested as a base for the CSF virus vaccine, plain sliced bread was considered the most appropriate base because of its absorbing ability and virus titre maintenance. Titres of bread‐based lyophilized CSF virus vaccine were stable at around 3.67 log₁₀ TCID₅₀ per ml for 7 months at 4°C. Pigs aged 5 and 8 weeks that orally received five bread‐based lyophilized CSF virus vaccine showed seroconversion of over 90% at 14 dpv. At 28 dpv, both age groups showed 100% seroconversion. In conclusion, the bread‐based lyophilized CSF virus vaccine can be an alternative choice for oral vaccination of pigs. Due to the simple process of production and the need for less virus titre, vaccine prices could be lowered. However, vaccine thermostability has to be improved to allow the vaccine delivery to be less dependent on functioning cool chains.     

Serological relationship between a novel ovine pestivirus and classical swine fever virus

The genus Pestivirus comprises globally distributed members of the family Flaviviridae, which cause severe losses in livestock. The most common species of the genus are bovine viral diarrhoea virus type 1 (BVDV‐1) and type 2 (BVDV‐2), classical swine fever virus (CSFV) and border disease virus (BDV). Recently, a novel ovine pestivirus was repeatedly detected in aborted lamb foetuses on a farm located in the Brescia Province (Italy). Complete genome characterization of this isolate showed that it was highly divergent from known pestivirus species and that it was genetically closely related to CSFV. The aim of this study was to determine the serological relatedness between the identified novel pestivirus and BVDV, BDV and CSFV selected strains for which homologous serum was available, by antigenic characterization performed using cross‐neutralization assays. The serological relatedness was expressed as the coefficient of antigenic similarity (R). Both field and specific antisera raised against the ovine pestivirus neutralized the CSFV reference strain Diepholz with titres significantly higher than those specific for the BDV and BVDV strains. Furthermore, the calculated R values clearly indicated that the novel ovine pestivirus is antigenically more related to CSFV than to ruminant pestiviruses, in agreement with the results of the genomic analysis. This would have severe consequences on CSFV serology in the event of a switch to porcine hosts with implications for CSFV surveillance and porcine health management.     

Revisiting the genetic diversity of classical swine fever virus: A proposal for new genotyping and subgenotyping schemes of classification

Classical swine fever (CSF ) is a highly contagious febrile viral disease caused by CSF virus (CSFV ), and it is considered one of the most important infectious diseases that affect domestic pigs and wild boar. Previous molecular epidemiology studies have revealed that the diversity of CSFV comprises three main genotypes and different subgenotypes defined using a reliable cut‐off to accurately classify CSFV at genotype and subgenotype levels. However, a growing number of CSFV both complete genome and full E2 gene sequences have been submitted to GenBank (more than 500 sequences are currently available, revised on December 1, 2017). Therefore, the aim of this study was to revisit the taxonomy of CSFV at genotype and subgenotype levels, to unify nomenclature and to provide an update to the classification of CSFV . We propose here a new genotyping scheme with five well‐defined CSFV genotypes (CSFV Genotypes 1–5) and 14 subgenotypes (seven for each of the CSFV Genotype 1 and CSFV Genotype 2). The findings showed in this study are relevant for molecular epidemiology approaches and will help to better understand the genetic diversity and spreading of CSFV at a global scale. The update in the classification of CSFV will allow the scientific community to establish more accurately the links among different outbreaks of the disease.     

Stability of classical swine fever virus and pseudorabies virus in animal feed ingredients exposed to transpacific shipping conditions

Classical swine fever virus (CSFV) and pseudorabies virus (PRV) are two of the most significant trade‐limiting pathogens affecting swine worldwide. Both viruses are endemic to China where millions of kilograms of feed ingredients are manufactured and subsequently imported into the United States. Although stability and oral transmission of both viruses through contaminated pork products has been demonstrated as a risk factor for transboundary spread, stability in animal feed ingredients had yet to be investigated. The objective of this study was to determine the survival of CSFV and variant PRV in 12 animal feeds and ingredients exposed to environmental conditions simulating a 37‐day transpacific shipment. Virus was detected by PCR, virus isolation and nursery pig bioassay. CSFV and PRV nucleic acids were stable throughout the 37‐day period in all feed matrices. Infectious CSFV was detected in two ingredients (conventional soybean meal and pork sausage casings) at 37 days post‐contamination, whereas infectious PRV was detected in nine ingredients (conventional and organic soybean meal, lysine, choline, vitamin D, moist cat and dog food, dry dog food and pork sausage casings). This study demonstrates the relative stability of CSFV and PRV in different feed ingredients under shipment conditions and provides evidence that feed ingredients may represent important risk factors for the transboundary spread of these viruses.     

The double‐antigen ELISA concept for early detection of Erns‐specific classical swine fever virus antibodies and application as an accompanying test for differentiation of infected from marker vaccinated animals

Emergency vaccination with live marker vaccines represents a promising control strategy for future classical swine fever (CSF) outbreaks, and the first live marker vaccine is available in Europe. Successful implementation is dependent on a reliable accompanying diagnostic assay that allows differentiation of infected from vaccinated animals (DIVA). As induction of a protective immune response relies on virus‐neutralizing antibodies against E2 protein of CSF virus (CSFV), the most promising DIVA strategy is based on detection of E^rns‐specific antibodies in infected swine. The aim of this study was to develop and to evaluate a novel E^rns‐specific prototype ELISA (pigtype CSFV E^rns Ab), which may be used for CSF diagnosis including application as an accompanying discriminatory test for CSFV marker vaccines. The concept of a double‐antigen ELISA was shown to be a solid strategy to detect E^rns‐specific antibodies against CSFV isolates of different genotypes (sensitivity: 93.5%; specificity: 99.7%). Furthermore, detection of early seroconversion is advantageous compared with a frequently used CSFV E2 antibody ELISA. Clear differences in reactivity between sera taken from infected animals and animals vaccinated with various marker vaccines were observed. In combination with the marker vaccine CP7_E2alf, the novel ELISA represents a sensitivity of 90.2% and a specificity of 93.8%. However, cross‐reactivity with antibodies against ruminant pestiviruses was observed. Interestingly, the majority of samples tested false‐positive in other E^rns‐based antibody ELISAs were identified correctly by the novel prototype E^rns ELISA and vice versa. In conclusion, the pigtype CSFV E^rns Ab ELISA can contribute to an improvement in routine CSFV antibody screening, particularly for analysis of sera taken at an early time point after infection and is applicable as a DIVA assay. An additional E^rns antibody assay is recommended for identification of false‐positive results in a pig herd immunized with the licensed CP7_E2alf marker vaccine.     

The evolution of African swine fever virus in Sardinia (1978–2014) as revealed by whole‐genome sequencing and comparative analysis

African swine fever (ASF) is a highly contagious and lethal viral disease of pigs and wild boars, which is enzootic in many African countries and on the Italian island of Sardinia, where it has been present since 1978. Previous genetic analyses of Sardinian ASF virus (ASFV) isolates have revealed that they all belong to p72 genotype I, with only minor sequence variations. However, these studies examined only a few selected genes. To distinguish between these closely related isolates and better investigate ASFV evolution in Sardinia, we sequenced the complete genomes of 12 Sardinian ASFV isolates collected between 1978 and 2012, and compared them with 47/Ss/2008 and 26544/OG10. Most of the observed changes occurred in a time‐dependent manner; however, their biological significance remains unclear. As a whole, our results demonstrate the remarkable genetic stability of these strains, supporting a single‐source introduction of the virus.     

Biological characterization of African swine fever virus genotype II strains from north‐eastern Estonia in European wild boar

Due to its impact on animal health and pig industry, African swine fever (ASF) is regarded as one of the most important viral diseases of pigs. Following the ongoing epidemic in the Transcaucasian countries and the Russian Federation, African swine fever virus was introduced into the Estonian wild boar population in 2014. Epidemiological investigations suggested two different introductions into the southern and the north‐eastern part of Estonia. Interestingly, outbreak characteristics varied considerably between the affected regions. While high mortality and mainly virus‐positive animals were observed in the southern region, mortality was low in the north‐eastern area. In the latter, clinically healthy, antibody‐positive animals were found in the hunting bag and detection of virus was rare. Two hypotheses could explain the different behaviour in the north‐east: (i) the frequency of antibody detections combined with the low mortality is the tail of an older, so far undetected epidemic wave coming from the east, or (ii) the virus in this region is attenuated and leads to a less severe clinical outcome. To explore the possibility of virus attenuation, a re‐isolated ASFV strain from the north‐eastern Ida‐Viru region was biologically characterized in European wild boar. Oronasal inoculation led to an acute and severe disease course in all animals with typical pathomorphological lesions. However, one animal recovered completely and was subsequently commingled with three sentinels of the same age class to assess disease transmission. By the end of the trial at 96 days post‐initial inoculation, all animals were completely healthy and neither virus nor viral genomes were detected in the sentinels or the survivor. The survivor, however, showed high antibody levels. In conclusion, the ASFV strain from north‐eastern Estonia was still highly virulent but nevertheless, one animal recovered completely. Under the experimental conditions, no transmission occurred from the survivor to susceptible sentinel pigs.     

A chemiluminescence immunoassay for rapid detection of classical swine fever virus E2 antibodies in pig serum samples

The high performance of chemiluminescence immunoassays (CLIAs) in diagnosis has been gradually recognized in recent years, but their application in the diagnosis of classical swine fever (CSF) has not been reported. Here, a recombinant E2 (rE2) protein and a peroxidase‐conjugated monoclonal antibody (MAb G5) were used to develop a competition‐based chemiluminescence immunoassay (cCLIA) for rapid and accurate detection of E2‐specific antibodies in pig serum. To evaluate the feasibility of cCLIA in the diagnosis of CSF, we developed a competition‐based enzyme‐linked immunosorbent assay (cELISA) as a control. Under the optimum test conditions, cCLIA showed a higher signal‐to‐noise ratio than that of the control cELISA. The best signal‐to‐noise ratios of cCLIA and cELISA were 70 and 17, respectively. Then, the diagnostic performance of the two assays was compared by examining a panel of pig serum samples (n = 285) with a confirmed status, and cCLIA showed higher diagnostic sensitivity (Dn) and diagnostic specificity (Dp) values than those of cELISA. The Dn and Dp of cCLIA were 97.49% and 96.08%, respectively, and those of cELISA were 93.97% and 94.12%, respectively. Furthermore, cCLIA can provide results within 20 min, whereas the control cELISA requires at least 1 hr. According to these findings, the newly developed cCLIA has potential application in the diagnosis of CSF and offers an alternative approach for efficient and rapid detection of E2‐specific antibodies.     

Comparative genomic analysis reveals an ‘open’ pan‐genome of African swine fever virus

The worldwide transmission of African swine fever virus (ASFV) drastically affects the pig industry and global trade. Development of vaccines is hindered by the lack of knowledge of the genomic characteristics of ASFV. In this study, we developed a pipeline for the de novo assembly of ASFV genome without virus isolation and purification. We then used a comparative genomics approach to systematically study 46 genomes of ASFVs to reveal the genomic characteristics. The analysis revealed that ASFV has an ‘open’ pan‐genome based on both protein‐coding genes and intergenic regions. Of the 151–174 genes found in the ASFV strains, only 86 were identified as core genes; the remainder were flexible accessory genes. Notably, 44 of the 86 core genes and 155 of the 324 accessory genes have been functionally annotated according to the known proteins. Interestingly, a dynamic number of taxis‐related genes were identified in the accessory genes, and two potential virulence genes were identified in all ASFV isolates. The ‘open’ pan‐genome of ASFV based on gene and intergenic regions reveals its pronounced natural diversity concerning genomic composition and regulation.     

Reduced specificity of Erns antibody ELISAs for samples from piglets with maternally derived antibodies induced by vaccination of sows with classical swine fever marker vaccine CP7_E2alf

Successful implementation of marker vaccines against classical swine fever virus is dependent on a reliable accompanying diagnostic assay that allows differentiation of infected from vaccinated animals (DIVA ) as well as the development of a testing scheme during emergency vaccination. In this context, special attention needs to be paid to breeding farms, because the offspring of marker vaccinated sows possess maternally derived antibodies (MDA s). So far, limited information is available on the influence of MDA s on serological testing in the context of a DIVA strategy. Therefore, two commercially available E^rns antibody ELISA s were compared, using serum samples of piglets with a high‐to‐moderate titre of MDA s against marker vaccine CP 7_E2alf. False‐positive results were detected by both E^rns antibody ELISA s for serum samples of piglets with an age of up to 4 weeks. Interestingly, most samples tested false‐positive in the first E^rns antibody ELISA were identified correctly by the other E^rns antibody ELISA and vice versa. In conclusion, in case of emergency vaccination of sows, the specificity of both ELISA s in newborn piglets younger than 4 weeks may be relatively low. This could be addressed in a testing strategy by either not sampling piglets up to the age of 4 weeks or using both ELISA s in a screening‐confirmation set‐up.     

Genome comparison of African swine fever virus China/2018/AnhuiXCGQ strain and related European p72 Genotype II strains

African swine fever was introduced into China in August 2018 and led to high mortality in domestic pigs. We reported the genome characterization of the China/2018/AnhuiXCGQ strain mainly based on next‐generation sequencing and comparison with related European p72 Genotype II strains. The genome was 189,393 bp long, encoding 181 open reading frames. Pair‐wise genome sequence comparison revealed 54–107 variation sites between China/2018/AnhuiXCGQ and the other genotype II virulent strains contributing to the change of expression or alteration of amino acid residues in 10–38 genes. China/2018/AnhuiXCGQ strain shared the highest similarity with POL/2015/Podlaskie strain. Phylogenetic analysis based on a 125 kb long conserved central region revealed that the China/2018/AnhuiXCGQ strain and four European genotype II strains were grouped into three clusters. This study expanded our knowledge on the genetic diversity and evolution of ASFV and provided valuable information for diagnosis improvement and vaccine development.     

Re‐emergence of genotype I of African swine fever virus in Ivory Coast

In July 2014, an outbreak of severe haemorrhagic disease in a domestic pig population, was reported in San‐Pedro, the second seaport city of Ivory Coast. Animals of all age groups developed clinical signs consistent with African swine fever (ASF). Tissue and serum samples from dead pigs were sent to the laboratory for diagnostic confirmation and molecular characterization based on the partial B646L (p72), the full E183L (p54) gene and the central variable region of the B602L gene. The PCR results confirmed the outbreak of ASF. Phylogenetic analyses based on p72 and p54 sequences showed that the San‐Pedro 2014 outbreak virus strain belongs to p72 genotype I. The Analysis of the tetrameric amino acid repeat regions of the B602L gene showed two repeat signatures which differ by an extra A = CAST in the second signature. The ASFV sequence of the San‐Pedro 2014 outbreak strain is closely related to historical and recent ASFV strains collected in Angola and Cameroon whose ships have repeatedly visited the seaport of San‐Pedro from March to June 2014. The 2014 viruses are distinct from the strains involved in the previous ASF wave in 1996 in Ivory Coast.     

Attenuated and non‐haemadsorbing (non‐HAD) genotype II African swine fever virus (ASFV) isolated in Europe,Latvia 2017

A non‐haemadsorbing (non‐HAD) ASF virus (ASFV) genotype II, namely Lv17/WB/Rie1, was isolated from a hunted wild boar in Latvia in 2017. Domestic pigs experimentally infected with the non‐HAD ASFV developed a nonspecific or subclinical form of the disease. Two months later, these animals were fully protected when exposed to other domestic pigs infected with a related virulent HAD genotype II ASFV.     

Positive selection pressure on E2 protein of classical swine fever virus drives variations in virulence,pathogenesis and antigenicity: Implication for epidemiological surveillance in endemic areas

Classical swine fever (CSF), caused by CSF virus (CSFV), is considered one of the most important infectious diseases with devasting consequences for the pig industry. Recent reports describe the emergence of new CSFV strains resulting from the action of positive selection pressure, due mainly to the bottleneck effect generated by ineffective vaccination. Even though a decrease in the genetic diversity of the positively selected CSFV strains has been observed by several research groups, there is little information about the effect of this selective force on the virulence degree, antigenicity and pathogenicity of this type of strains. Hence, the aim of the current study was to determine the effect of the positive selection pressure on these three parameters of CSFV strains, emerged as result of the bottleneck effects induced by improper vaccination in a CSF‐endemic area. Moreover, the effect of the positively selected strains on the epidemiological surveillance system was assessed. By the combination of in vitro, in vivo and immunoinformatic approaches, we revealed that the action of the positive selection pressure induces a decrease in virulence and alteration in pathogenicity and antigenicity. However, we also noted that the evolutionary process of CSFV, especially in segregated microenvironments, could contribute to the gain‐fitness event, restoring the highly virulent pattern of the circulating strains. Besides, we denoted that the presence of low virulent strains selected by bottleneck effect after inefficient vaccination can lead to a relevant challenge for the epidemiological surveillance of CSF, contributing to under‐reports of the disease, favouring the perpetuation of the virus in the field. In this study, B‐cell and CTL epitopes on the E2 3D‐structure model were also identified. Thus, the current study provides novel and significant insights into variation in virulence, pathogenesis and antigenicity experienced by CSFV strains after the positive selection pressure effect.     

No evidence for long‐term carrier status of pigs after African swine fever virus infection

This study targeted the assessment of a potential African swine fever virus (ASFV) carrier state of 30 pigs in total which were allowed to recover from infection with ASFV “Netherlands'86” prior exposure to six healthy sentinel pigs for more than 2 months. Throughout the whole trial, blood and swab samples were subjected to routine virological and serological investigations. At the end of the trial, necropsy of all animals was performed and viral persistence and distribution were assessed. Upon infection, a wide range of clinical and pathomorphological signs were observed. After an initial acute phase in all experimentally inoculated pigs, 66.6% recovered completely and seroconverted. However, viral genome was detectable in blood samples for up to 91 days. Lethal outcomes were observed in 33.3% of the pigs with both acute and prolonged courses. No ASFV transmission occurred over the whole in‐contact phase from survivors to sentinels. Similarly, infectious ASFV was not detected in any of the tissue samples from ASFV convalescent and in‐contact pigs. These findings indicate that the suggested role of ASFV survivors is overestimated and has to be reconsidered thoroughly for future risk assessments.     

Genetic characterization of African swine fever virus isolates from soft ticks at the wildlife/domestic interface in Mozambique and identification of a novel genotype

African swine fever virus (ASFV ) is one of the most threatening infectious diseases of pigs. There are not sufficient data to indicate the importance of the sylvatic cycle in the spread and maintenance of the disease locally and potentially, globally. To assess the capacity to maintain ASF in the environment, we investigated the presence of soft tickreservoirs of ASFV in Gorongosa National Park (GNP ) and its surrounding villages. A total of 1,658 soft ticks were recovered from warthog burrows and pig pens at the wildlife/livestock interface of the GNP and viral DNA was confirmed by nested PCR in 19% of Ornithodoros porcinus porcinus and 15% of O. p. domesticus . However, isolation of ASFV was only achieved in approximately 50% of the PCR ‐positive samples with nineteen haemadsorbing virus isolates recovered. These were genotyped using a combination of partial sequencing of the B646L gene (p72 ) and analysis of the central variable region (CVR ) of the B602L gene. Eleven isolates were classified as belonging to genotype II and homologous to contemporary isolates from southern Africa, the Indian Ocean and eastern Europe. Three isolates grouped within genotype V and were similar to previous isolates from Mozambique and Malawi. The remaining five isolates constituted a new, previously unidentified genotype, designated genotype XXIV . This work confirms for the first time that the virus currently circulating in eastern Europe is likely to have a wildlife origin, and that the large diversity of ASFV maintained in wildlife areas can act as a permanent sources of different strains for the domestic pig value chain in Mozambique and beyond its boundaries. Their genetic similarity to ASFV strains currently spreading across Europe justifies the need to continue studying the sylvatic cycle in this African country and other parts of southern Africa in order to identify potential hot spots of ASF emergence and target surveillance and control efforts.     

A modified‐live porcine reproductive and respiratory syndrome virus (PRRSV)‐1 vaccine protects late‐term pregnancy gilts against heterologous PRRSV‐1 but not PRRSV‐2 challenge

The objective of this study was to determine the efficacy of a commercially available porcine reproductive and respiratory syndrome virus (PRRSV )‐1 modified‐live virus (MLV ) vaccine against PRRSV ‐1 and PRRSV ‐2 challenge in late‐term pregnancy gilts. Gilts were vaccinated with the PRRSV ‐1 MLV vaccine at 4 weeks prior to breeding and then challenged intranasally with PRRSV ‐1 or PRRSV ‐2 at 93 days of gestation. After PRRSV ‐1 challenge, vaccinated pregnant gilts had a significantly longer gestation period, significantly higher numbers of live‐born and weaned piglets and a significantly lower number of stillborn piglets at birth compared to unvaccinated pregnant gilts. No significant improvement in reproductive performance was observed between vaccinated and unvaccinated pregnant gilts following PRRSV ‐2 challenge. Vaccinated pregnant gilts also exhibited a significantly improved reproductive performance after challenge with PRRSV ‐1 compared to vaccinated pregnant gilts following PRRSV ‐2 challenge. The PRRSV ‐1 MLV vaccine was able to reduce PRRSV ‐1 but not PRRSV ‐2 viremia in pregnant gilts. Vaccinated gilts also showed a significantly higher number of PRRSV ‐1‐specific IFN ‐γ‐secreting cells (IFN ‐γ‐SC ) compared to PRRSV ‐2‐specific IFN ‐γ‐SC . The data presented here suggest that the vaccination of pregnant gilts with a PRRSV ‐1 MLV vaccine provides good protection against PRRSV ‐1 but only limited protection against PRRSV ‐2 challenge in late‐term pregnancy gilts based on improvement of reproductive performance, reduction in viremia and induction of IFN ‐γ‐SC .     

Overcoming the challenges of pen‐side molecular diagnosis of African swine fever to support outbreak investigations under field conditions

African swine fever (ASF) is a devastating disease of pigs. Without a vaccine, early detection and rapid diagnosis of ASF is a crucial step towards effective disease control. In many countries where ASF is endemic, laboratory infrastructure including sampling and sample shipment is inadequate, and a rapid laboratory confirmation would require that the diagnosis is performed at regional laboratories close to the pig farms of concern, or even at the farm‐side. This study intended to evaluate measures including sample preparation methods, a dried‐down assay, and a portable, battery‐powered real‐time PCR instrument, to improve molecular diagnosis under field conditions. A simple dilution of blood samples, either in Phosphate‐buffered saline or a commercial buffer, worked similarly to beads‐based nucleic acid extraction using a magnet as the core equipment; the latter method did work as well for those samples with low viral load or high Ct values. The real‐time PCR assay using a Universal ProbeLibrary (UPL) probe tolerated suspected inhibitory substances present in the prepared samples better, whereas the dried‐down assay had a higher diagnostic sensitivity. Additionally, an inhibition control assay proved to be helpful in avoiding false negative results when interpreting negative results of samples that might be of low quality or with inadequate reduction in inhibitory substances. When tested with synthetic DNA standards, the portable instrument performed at a level approaching stationary thermocyclers. In summary, the developments of suitable sample preparation methods, robust and thermal‐stable real‐time PCR assays with inhibition control, and battery‐powered portable thermocyclers with middle‐throughput offer one way forward to provide rapid, reliable molecular diagnosis under challenging field conditions.     

The infectious cDNA clone of commercial HP‐PRRS JXA1‐R‐attenuated vaccine can be a potential effective live vaccine vector

Multiple commercial porcine reproductive and respiratory syndrome (PRRS) modified live vaccines are currently utilized in Chinese swine herds due to the limited cross‐protection of vaccines and coexistence of different PRRS viruses. In this study, an infectious cDNA clone of the highly pathogenic PRRS (HP‐PRRS) vaccine JXA1‐R strain was generated. We successfully rescued the virus from direct in vitro DNA transfection of rJXA1‐R clone, which has similar growth kinetics to the parental JXA1‐R virus in Marc‐145 cells. To further evaluate the potential use of the cloned rJXA1‐R virus as a live vector for foreign gene expression, the enhanced green fluorescent protein (EGFP) was inserted between non‐structural and structural genes. Our results showed that the dynamic expression of EGFP can be visualized by live cell imaging system during the infection in Marc‐145 cells. The availability of our cloned JXA1‐R viruses provides a crucial platform to study the fundamental biology of HP‐PRRS virus vaccine and also serves as a potential effective vector for developing live vector vaccines against swine pathogens.     

Diagnostic specificity of the African swine fever virus antibody detection enzyme‐linked immunosorbent assay in feral and domestic pigs in the United States

African swine fever (ASF) is a highly contagious haemorrhagic disease of pigs that has the potential to cause mortality nearing 100% in naïve animals. While an outbreak of ASF in the United States’ pig population (domestic and feral) has never been reported, an introduction of the disease has the potential to cause devastation to the pork industry and food security. During the recovery phase of an outbreak, an antibody detection diagnostic assay would be required to prove freedom of disease within the previously infected zone and eventually nationwide. Animals surviving an ASF infection would be considered carriers and could be identified through the persistence of ASF viral antibodies. These antibodies would demonstrate exposure to the disease and not vaccination, as there is no ASF vaccine available. A well‐established commercial enzyme‐linked immunosorbent assay (ELISA) detects antibodies against ASF virus (ASFV), but the diagnostic specificity of the assay had not been determined using serum samples from the pig population of the United States. This study describes an evaluation of the World Organization for Animal Health (OIE)‐recommended Ingezim PPA COMPAC ELISA using a comprehensive cohort (n = 1791) of samples collected in the United States. The diagnostic specificity of the assay was determined to be 99.4% (95% confidence interval (CI): [98.9, 99.7]). The result of this study fills a gap in understanding the performance of the Ingezim PPA COMPAC ELISA in the ASF naïve pig population of the United States.