Search Results (341)

Porcine reproductive and respiratory syndrome virus (PRRSV), an enveloped single-stranded positive-sense RNA virus, poses a significant threat to global swine production. Despite the availability of modified live virus and inactivated vaccines, their limited efficacy and safety concerns highlight the urgent need for novel antiviral therapeutics. This study aimed to investigate the molecular mechanisms by which lycopene inhibits PRRSV replication. Initial assessments confirmed that lycopene did not adversely affect cellular viability, cell cycle progression, or apoptosis. Using fluorescence microscopy, flow cytometry, immunoblotting, quantitative real-time PCR (qRT-PCR), and viral titration assays, lycopene was shown to exhibit potent antiviral activity against PRRSV. Mechanistic studies revealed that lycopene suppresses reactive oxygen species (ROS) production, which is critical for PRRSV proliferation. Additionally, lycopene attenuated PRRSV-induced inflammatory responses, as demonstrated by immunoblotting, ELISA, and qRT-PCR assays. These findings suggest that lycopene inhibits PRRSV replication by modulating ROS levels and mitigating inflammation, offering a promising avenue for the development of antiviral therapeutics. This study provides new insights and strategies for combating PRRSV infections, emphasizing the potential of lycopene as a safe and effective antiviral agent. Full article

Porcine reproductive and respiratory syndrome virus remains one of the most economically significant pathogens in swine production, with PRRSV-2 being the dominant variant in the United States. While lineage classification has traditionally relied on ORF5 sequencing, recent studies suggest that this single-gene approach may overlook key evolutionary events such as recombination. In this study, we performed whole-genome sequencing and phylogenetic analysis of seven PRRSV-2 isolates collected in the U.S. between 2006 and 2024. Using reference-guided assembly, lineage assignment, and recombination detection with RDP5 and SIMplot, we identified discordant phylogenetic placements between ORF5 and whole genomes in four of the seven isolates. These discordances were explained by multiple recombination events affecting different genomic regions, particularly ORF2–ORF7. In contrast, three isolates showed phylogenetic concordance and no strong evidence of recombination. Our findings demonstrate that recombination plays a significant role in shaping PRRSV-2 evolution and highlight the limitations of ORF5-based lineage classification. Whole-genome surveillance is therefore essential to accurately track viral diversity, detect recombinant strains, and inform control strategies. This work underscores the need for a broader adoption of full-genome analysis in routine PRRSV surveillance and research. Full article

The seasonal patterns of viral diseases in farm animals present significant challenges to global livestock productivity, with cold stress emerging as a potential modulator of host–pathogen interactions. This review synthesizes current knowledge on the expression dynamics of heat shock protein 70 (HSP70) in farm animals under cold-stress conditions and its potential roles as (1) a viral replication facilitator and (2) an immune response regulator. This review highlights cold-induced HSP70 overexpression in essential organs, as well as its effects on significant virus life cycles, such as porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV), and bovine viral diarrhea virus (BVDV), through processes like viral protein chaperoning, replication complex stabilization, and host defense modulation. By integrating insights from thermophysiology, virology, and immunology, we suggest that HSP70 serves as a crucial link between environmental stress and viral disease seasonality. We also discuss translational opportunities targeting HSP70 pathways to break the cycle of seasonal outbreaks, while addressing key knowledge gaps requiring further investigation. This article provides a framework for understanding climate-driven disease patterns and developing seasonally adjusted intervention strategies. Full article

Interferon-induced transmembrane protein 3 (IFITM3) is a member of the family of interferon-stimulated genes (ISGs) that inhibits a diverse array of enveloped viruses which enter host cells by endocytosis. Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped RNA virus causing significant economic losses to the swine industry. Very little is known regarding how IFITM3 restricts PRRSV. In this study, the role of IFITM3 in PRRSV infection was studied in vitro using MARC-145 cells. IFITM3 over-expression reduced PRRSV replication, while the siRNA-induced knockdown of endogenous IFITM3 increased PRRSV RNA copies and virus titers. The colocalization of the virus with IFITM3 was observed at both 3 and 24 h post infection (hpi). Quantitative analysis of confocal microscopic images showed that an average of 73% of IFITM3-expressing cells were stained positive for PRRSV at 3 hpi, while only an average of 27% of IFITM3-expressing cells were stained positive for PRRSV at 24 hpi. These findings suggest that IFITM3 may restrict PRRSV at the post-entry steps. Future studies are needed to better understand the mechanisms by which this restriction factor inhibits PRRSV. Full article

Since its emergence in China in 2018, African swine fever virus (ASFV) has posed a severe threat to the pig farming industry due to its high transmissibility and mortality rate. The clinical signs of ASFV infection often overlap with those caused by other swine viruses such as classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), and porcine circovirus type 2 (PCV2), making timely and precise diagnosis a considerable challenge. To address this, we established a TaqMan-based multiplex real-time quantitative PCR (qPCR) assay capable of simultaneously detecting ASFV, CSFV, PRRSV, PRV, and PCV2. Specific primer-probe sets were developed targeting conserved genomic regions: the ASFV P72 gene, CSFV 5’UTR region, PRRSV ORF6, PCV2 cap gene, and PRV gB gene. After thorough optimization, the assay demonstrated robust analytical performance, exhibiting strong target specificity with no cross-detection of non-target pathogens. The detection threshold was determined to be 10 copies/μL per virus, indicating high assay sensitivity. Repeatability analysis revealed low variability, with intra- and inter-assay coefficient of variation values remaining below 2.3%. When applied to 95 clinical samples, the multiplex assay yielded results that were fully consistent with those obtained using commercially available singleplex qPCR kits. In conclusion, the multiplex TaqMan qPCR method developed in this study is characterized by high specificity, sensitivity, and reproducibility. It provides a reliable and efficient diagnostic tool for the simultaneous detection and differential diagnosis of ASFV and other clinically similar viral infections in swine, thereby offering robust technical support for swine disease surveillance and control. Full article

Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2) represents one of the greatest threats to global pork production. Increased incidence of a genetic variant of Porcine Reproductive and Respiratory Syndrome Virus (variant 1H.18) was recently reported in the Midwestern USA Sequence comparisons in the ORF5 region indicate that 1H.18 was closely related to both sub-lineages L1H and L1C. To expand our understanding and attempt to elucidate the origin of the 1H.18, we sequenced a near-complete genome, covering all coding regions, and investigated the occurrence of recombination events that may have contributed to the emergence of the new variant. At least six distinct recombination events were identified across the coding portion of the genome. Evidence of recombination in the ORF5 region between variants 1H.31 and 1C.3 was detected. Our results suggest a likely origin of 1H.18 driven by recombination. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major viral threat to swine, causing significant economic loss in the global pig farming industry. This virus includes two major genotypes, PRRSV1 and PRRSV2, both characterized by high mutation rates and genetic variability, complicating the development of a universally effective vaccine and disease control. To address this challenge, this study utilizes immunoinformatics tools to identify conserved epitopes and design a multi-epitope vaccine candidate against PRRSV based on reverse vaccinology. The complete sequences of PRRSV-encoded proteins were retrieved worldwide, and the conserved fragments were identified through the alignment of polypeptide sequences. Subsequent screening was conducted to screen epitopes for their potential to be safe and to activate B cells, HTLs (helper T cells), and CTLs (cytotoxic T cells). By conjugating the selected epitopes with distinct adjuvant proteins, three vaccine candidates were designed and termed PRRSV-vaccine (PRRSV-V-1, PRRSV-V-2, and PRRSV-V-3, respectively). Furthermore, systematic evaluations of their physicochemical properties, structural stability, binding with pattern recognition receptors, and induction of the host immune system were performed. PRRSV-V-2 had the most promising physicochemical and structural characteristics, strong binding with toll-like receptors (TLR3 and TLR8), and the most vigorous reactions to host immune responses. As the most promising candidate, the recombinant PRRSV plasmid was in silico designed for expression in Escherichia coli. Our study proposed a novel approach to PRRSV vaccine development against PRRSV, offering a promising strategy for controlling the infection across diverse PRRSV strains in swine. Despite providing significant insights into vaccine design through computational methods, the results of this study remain predictive. So, it is open for the experimental validations of the scientific community to ensure its actual immunological properties, especially the safety and efficacy. Full article

Porcine circovirus type 2 (PCV2) is the main cause of porcine circovirus-associated disease (PCVAD). Despite the widespread use of anti-PCV2 vaccines, their efficacy varies, influenced by co-infection and evaluation methods. This study assessed the efficacy of Ingelvac CircoFLEX^® PCV2 vaccine under natural conditions. One hundred serum samples were collected from vaccinated and non-vaccinated piglets aged 21 to 173 days. PCR and antibody positivity rates did not show significant differences between the two groups, but PCV2 gene load at 91 days was significantly lower (p = 0.0095) in the vaccinated group. Anti-PCV2 antibody titers were also significantly lower in the vaccinated group at 91, 145, and 173 days (p < 0.0001). PCV2 was isolated from 50% of piglets in the non-vaccinated group (50%), compared with none (0%) in the vaccinated group, suggesting that PCV2 gene load in the non-vaccinated group did not correlate with viremia. Both groups were positive for antibodies to porcine reproductive and respiratory syndrome virus (PRRSV) at 63 days, prior to the surge in PCV2 gene load, suggesting PRRSV may enhance PCV2 replication. These findings highlight that while the vaccine reduced PCVAD damage, evaluation should incorporate methods such as virus isolation instead of relying solely on PCR. Full article

Porcine reproductive and respiratory syndrome (PRRS) is aworldwide spread disease. This study analyzed the changes in saliva analytes of pigs infected with PRRS virus (PRRSV) in different clinical conditions that can appear in PRRSV-positive farms. Biomarkers for inflammation (haptoglobin, total proteins), immune response (adenosine deaminase), tissue damage (lactate dehydrogenase), stress (alpha-amylase), and sepsis (calprotectin, aldolase, Serpin B12) were measured in pigs under three clinical scenarios: (1) no evident clinical signs, (2) clinical signs indicating PRRSV activation, and (3) secondary bacterial infection by Streptococcus suis. Haptoglobin and lactate dehydrogenase showed significant increases in pigs with PRRSV activation compared to pigs without clinical signs. Additionally, the levels of Serpin B12, aldolase, calprotectin, total proteins, and the activity of adenosine deaminase significantly increased in pigs with meningitis compared to pigs without clinical signs, but did not show significant differences between healthy pigs and those with PRRSV clinical signs without bacterial infection. In summary, PRRSV-infected pigs can show differences in selected saliva analytes depending on their clinical condition. These findings may have practical applications for detecting PRRSV infections and differentiating cases with associated meningitis. Full article

The global pork production sector continues to experience substantial financial burdens attributable to porcine reproductive and respiratory syndrome virus (PRRSV) infections. Despite the current epidemiological landscape in which NADC30-like strains predominate alongside cocirculating diverse PRRSV subtypes, highly pathogenic PRRSV (HP-PRRSV) remains a persistent threat. Furthermore, currently available commercial PRRS vaccine formulations exhibit restricted heterologous protection efficacy. The development of novel mRNA-based vaccines represents a promising strategy for PRRS mitigation protocols. In response to these epidemiological challenges, an HP-PRRSV strain (Lineage 8), designated as JX021, was isolated and characterized in this study. Pathogenicity experiments confirmed that JX021 induces severe clinical symptoms in piglets. Moreover, by combining immunoinformatics and literature-guided approaches, critical antigenic epitopes on HP-PRRSV (represented by the JXA1 strain) structural proteins were identified, enabling the design and synthesis of a multiepitope mRNA vaccine. The survival of piglets immunized with the mRNA vaccine was higher than that of the inactivated vaccine immunization group and the PBS group. Compared with the inactivated vaccine group, the mRNA vaccine group presented reductions in viremia and lung lesions. These findings provide new insights into the design and development of further PRRS vaccine research. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV) poses a significant economic challenge to Japan’s swine industry. This review synthesizes the epidemiological evolution of PRRSV in Japan by examining the available scientific literature from its initial Type 2 isolation in 1993 to recent events. Endemic Type 2 strains, initially dominated by Cluster III (Lineage 4), have diversified significantly. This diversification was marked by key events including the emergence of vaccine-associated Cluster II (Lineage 5) and incursions of virulent Cluster IV/Lineage 1F (MN184A-like) strains and Type 1 virus around 2008. By 2018–2020, Clusters II and IV predominated nationwide, a trend strongly linked to widespread modified live virus (MLV) vaccination. The recent detection of the globally significant NADC34-like (Lineage 1A) strain underscores ongoing foreign incursion risks. Current MLV vaccines face challenges, including safety concerns and limited cross-protection against diverse field strains. Consequently, effective control requires integrated strategies, comprising optimized vaccination, stringent biosecurity, advanced molecular surveillance, improved diagnostics, and coordinated regional control programs guided by systematic herd classification and stakeholder partnerships. The development of next-generation vaccines and sustained multi-stakeholder collaboration are critical for mitigating the impact of PRRSV in Japan. Full article

Due to its high genomic variability, the epidemiological landscape of porcine reproductive and respiratory syndrome virus (PRRSV) has become increasingly complex in recent years. From 2022 to 2023, we collected a total of 1044 clinical samples from pigs suspected of PRRSV infection in China and discovered a PRRSV-positive rate of 29.8% (311/1044) using RT-PCR targeting the nsp2 gene. Among these positive samples, NADC30/34-like PRRSV, highly pathogenic PRRSV (HP-PRRSV), and classical PRRSV strains accounted for 60.1%, 37.9%, and 4.5%, respectively. These results indicate that the most prevalent PRRSV strains in China are NADC30/34-like PRRSV, followed by HP-PRRSV. Two PRRSV strains, JX03 and HN08, were isolated, and TCID₅₀ assays were performed to determine their titers at different time points post-infection, revealing differences in their proliferation kinetics. Phylogenetic, amino acid sequence, and recombination analyses demonstrated that the JX03 and HN08 strains cluster within lineage 8 (HP-PRRSV) and sublineage 1.5 (NADC34-like PRRSV), respectively. Notably, the HN08 strain was identified as a recombinant between the NADC30-like and NADC34-like strains, while no recombination event was detected in the JX03 strain. Pathogenicity assessments showed that the JX03 strain exhibited higher pathogenicity than the CHN-HB-2018 strain (a NADC30-like PRRSV strain was previously isolated by our lab), as evidenced by differences in clinical signs and mortality rates in piglets. In contrast, HN08 displayed no obvious clinical symptoms or mortality, revealing lower pathogenicity than the CHN-HB-2018 strain. These findings provide valuable information on the epidemiological and genetic characteristics of PRRSV strains in China, laying a foundation for the development of effective strategies against PRRSV. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV) modified live virus (MLV) vaccination is used to control PRRSV. In China, farms conduct random sampling from sow herds every 4 to 6 months. They use the enzyme-linked immunosorbent assay (ELISA) method to monitor the immune status of the herd by tracking the positive rate or the sample-to-positive ratio. However, in farms that implement mass vaccination and have stable production, the positive rate of ELISA antibodies has decreased, especially in high-parity sows. This poses a considerable challenge to the current monitoring approach of PRRSV immunity. It remains unclear whether this reflects insufficient sensitivity of the kits for these special scenarios or the fact that the sows have truly lost immunity. In this study, 233 samples from four farms (A–D) across different regions of China were acquired. They were tested using four representative ELISA kits, two targeting the nucleocapsid protein (N) and two targeting the glycoprotein (GP) to evaluate PRRS immune status. The respective sample positive rates in A–D were 57.1–100%, 50.9–100%, 50–100%, and 75.7–100% using the kits. The positive rates using the four ELISA kits were 50.0–75.7%, 70.0–75.7%, 82.5–97.1%, and 100%, respectively, with poor agreement among them. The positive rates and humoral antibody levels for parity 1 and 2 sows were significantly lower than those with higher parities (>4). Eighty-eight ELISA-negative samples identified using ELISA kit A were verified using a viral neutralizing test (VNT), with only 15.9% of the samples testing negative. In conclusion, the ELISA antibody negativity issue existed, mostly occurring in specific farms tested using a specific kit. However, the low correlation with the VNT results and the poor agreements among the kits suggest that relying on one ELISA test is insufficient to monitor the immune status of PRRSV MLV-vaccinated herds. Full article

First reported in 1987, the porcine reproductive and respiratory syndrome virus (PRRSV) has significantly disrupted the major regions affected by PRRSV in the pig breeding industry. Recently, outbreaks of disease caused by recombinant PRRSV strains in China have raised serious concerns. Effective immunization and infection control in pig populations is critical, as the virus frequently undergoes mutation and recombination. This study characterized a novel recombinant PRRSV strain, BX/CH/22, isolated from Northeast China. Genetic analysis revealed that BX/CH/22 is a recombinant of JXA1, NADC 30-like, and NADC 34-like strains. Phylogenetic analysis of the non-structural protein (NSP) 2 region classified BX/CH/22 as JXA1 PRRSV-like, with a characteristic deletion of 30 discontinuous amino acids in NSP2. However, Open Reading Frame (ORF) 5 analysis classified it as NADC 30-like PPRSV, while whole-genome phylogenetic analysis classified it as NADC 34-like PPRSV. Recombination analysis revealed that BX/CH/22 contains an NADC 34-like PRRSV backbone, an NSP-coding region from NADC 30-like PRRSV, and an ORF2-ORF6 region from NADC 34-like PRRSV. The strain was isolated from serum samples obtained from commercial swine farms undergoing active PRRS outbreaks. In animal experiments, all BX/CH/22-challenged piglets exhibited persistent fever, with peak temperatures >40.5 °C at 4–9 dpi resolving by 11 dpi, accompanied by cough, anorexia, and lethargy. A significant reduction in daily weight gain was observed in infected groups compared to asymptomatic controls, with a 100% survival rate. Our findings provide early warning for PRRSV immune control strategies. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV) is considered a promising viral vector for the expression and delivery of foreign genes for the development of a new generation of multi-valent vaccines against PRRSV and other porcine viruses, as well as for analyses of the immune response against PRRSV and anti-PRRSV component screening. In the present study, the junction site between nsp1β and nsp2 in the PRRSV genome was tested for the insertion and expression of foreign genes. Three foreign genes, including eGFP, iLOV3, and TEVp, were inserted into the intergenic junction between nsp1β and nsp2 and expressed by the respective recombinant PRRSVs (rPRRSV-SH01-eGFP, rPRRSV-SH01-iLOV3, and rPRRSV-SH01-TEVp) in vitro in mammalian cells. Analysis of the growth kinetics of the rescued recombinant PRRSVs showed no significant differences between the recombinant PRRSVs and their parental viruses. The inserted genes were consistently present in the viral genome during serial passage in vitro (for at least 20 passages). In addition, rPRRSV-SH01-eGFP can be used as a reporter virus for rapid detection of neutralizing antibodies against PRRSV through a fluorescent focus unit reduction-based assay. These data demonstrate that the junction between nsp1β and nsp2 is a new site that is suitable for the insertion and expression of foreign genes, providing a new option to express and deliver foreign genes using PRRSV-based vectors for different purposes, such as the development of multi-valent vaccines against PRRSV and other porcine viruses. Full article