Search Results (97)

The distribution and genetic diversity of economically significant pathogens, including porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), porcine circovirus type 3 (PCV3), and porcine parvovirus 1 (PPV1), across extensive Russian territory within wild boars that serve as reservoirs remain poorly characterized. This study aimed to conduct a molecular epidemiological survey of these viruses in wild boar populations. The samples of 476 wild boars, collected across Russia between 2021 and 2025, were tested by PCR for the detection of viral genomes. While PRRSV was not detected, we found high detection rates for PCV2 (34.9%), PCV3 (35.5%), and PPV1 (25.4%). For PCV2, the co-circulation of two genotypes, PCV2b (5/53) and PCV2d (48/53), was observed. All 29 PCV3 sequences belonged to the PCV3a genotype. For PPV1, the presence of the PPV1a, PPV1b, and PPV1d genotype, as well as unclassified isolates, was shown. Co-infection of different viruses was detected: PCV2/PCV3 (16.0%), PCV2/PPV1 (6.9%), PCV2/PCV3/PPV1 (6.9%), and PCV3/PPV1 (4.4%). This is the first comprehensive study that demonstrates the wide dissemination and genetic diversity of PCV2, PCV3, and PPV1 within the wild boar population in Russia and highlights their role as a potential reservoir in viral evolution and spread. Full article

Porcine circovirus 3 (PCV3), first reported in 2016, is associated with diverse clinical conditions, including porcine dermatitis and nephropathy syndrome, reproductive disorders, and systemic inflammation, and affects pigs of all ages. To investigate the prevalence and genetic evolution of PCV3 in Hunan Province, China, 700 lymph node tissue specimens were collected from slaughterhouses and hazard-free disposal centers across 14 prefecture-level cities between 2021 and 2024 and screened using real-time quantitative PCR (qPCR). Epidemiological investigation revealed an overall PCV3 positivity rate of 29.4% (206/700) in the province. The highest prevalence was observed in Yiyang City (56%, 28/50), whereas no positive samples were detected in Zhuzhou City (0/30). Among the positive samples, 34 specimens from different cities with Ct values < 25 were selected for Cap gene amplification and sequencing. Phylogenetic analysis showed that PCV3c was the predominant genotype (67.6%, 23/34), followed by PCV3a (32.4%, 11/34), while PCV3b was not detected. We identified twelve amino acid substitution sites within Cap proteins. Furthermore, B-cell linear epitope prediction and homology modeling of the Cap protein identified seven linear epitopes, with ten amino acid variation sites located within these epitopic regions. This study enriches the molecular epidemiological data of PCV3 in southern China and provides a reference for future PCV3 control strategies. Full article

Porcine circovirus 2 (PCV2) is a DNA virus that is classified in the genus Circovirus of the Circoviridae family, which is a causative agent of Porcine Circovirus-Associated disease (PCVAD). PCVAD continues to cause substantial losses in global pig farming, with PCV2d being the prevalent genotype worldwide, including in Vietnam. In this study, we focused on generating a recombinant PCV2d Cap protein fused to the GCN4pII motif (Cap2d-pII) in a plant-based system and evaluating its immunogenicity. The Cap2d-pII gene was cloned into a plant expression vector and introduced into Agrobacterium tumefaciens for transient expression in Nicotiana benthamiana leaves. Western blot analysis confirmed the high accumulation of the Cap2d-pII protein, which was purified by Immobilized affinity chromatography and used for immunizing mice. ELISA and immunoperoxidase monolayer assay results demonstrated that immunization with the recombinant protein elicited robust humoral and cellular immune responses. At 56 days after immunization, mice vaccinated with the Cap2d-pII protein generated PCV2d-specific IgG titers and IFN-γ responses that were consistent with those in mice receiving the commercial inactivated vaccine. These observations confirm that the plant-expressed Cap2d-pII antigen effectively activates both antibody- and T cell-mediated immune pathways. Collectively, this study identifies the Cap2d-pII protein as a promising plant-derived vaccine candidate for the development of effective and affordable PCV2d subunit vaccines. Full article

Background: Porcine circovirus type 2 (PCV2) remains one of the most important pathogens that infects swine, causing considerable economic losses worldwide. PCV2 vaccines are commercially available, and the development of experimental vaccines that could confer better protection against emerging genotypes is underway. The expression of virus-like particles (VLPs) carrying different PCV2 capsid (Cap) peptides in E. coli was recently reported. These chimeric particles were adjuvated with an oil-in-water emulsion with polymer and induced different titers of serum IgG in BALB/c mice after a single subcutaneous injection. The aim of this study was to assess the immune response and protective efficacy elicited by VLPs carrying the PCV2b Cap carboxy-terminal peptide in the target species. Methods: Domestic pigs (Sus scrofa domesticus) were immunized intramuscularly with 25 μg of adjuvated chimeric VLPs on days 0 and 14 and challenged on day 28 with a PCV2b Mexican isolate. PCV2 peptide-specific IgG seroconversion, serum cytokines, viral load in nasal swabs and organs, and histopathological score were determined. Results: IgG levels peaked 28 days post-immunization. Interleukin-12 and -18 and interferon-gamma increased 21 days after immunization. In addition, genomic material of PCV2 was detected in nasal swabs from one specimen on day 7, two specimens on day 14, and two specimens on day 21 following viral challenge. Finally, histological lesions were not less severe in immunized specimens compared to non-vaccinated/challenged specimens. Conclusions: These results suggest that immunization with chimeric VLPs could contribute to controlling viral shedding in pig herds where a PCV2b genotype is most prevalent. Full article

Porcine circovirus types 2 (PCV2) and 3 (PCV3) are major pathogens affecting swine health and productivity, yet important gaps remain in understanding their evolution and circulation in Europe, particularly within wild boar populations that may serve as reservoirs. This study examined the genetic diversity and evolutionary dynamics of PCV2 and PCV3 in Portugal, drawing on viral genomes obtained from domestic pigs and wild boars to explore transmission patterns, spillover events and the contribution of recombination to viral emergence. We identified two PCV2 genotypes (PCV2a and PCV2d) and two PCV3 genotypes (PCV3-2a and PCV3-3g) circulating in Portuguese swine. Phylogeographic reconstruction revealed multiple introductions of both PCV2 and PCV3 from China into Europe, followed by regional diversification and subsequent spread within European wild boar populations. Evidence of bidirectional viral exchange between domestic pigs and wild boars was also observed. Recombination played a notable role in PCV2 evolution, with consistent signals detected among PCV2a sequences and indications that the PCV2h genotype likely originated from a recombinant event involving a Portuguese PCV2a strain and a Chinese PCV2d strain. By contrast, no recombination was detected in PCV3, suggesting that its evolution is primarily mutation-driven. Overall, these findings highlight the complex evolutionary history of swine circoviruses in Europe and underscore the importance of continuous genomic surveillance in both domestic and wild hosts. The study reinforces the value of a One Health approach for monitoring and controlling emerging circoviruses with implications for animal health and livestock production. Full article

Background/Objectives: Maternally derived antibody (MDA) levels of porcine circovirus 2 (PCV2) may eventually interfere with humoral response and vaccination efficacy. This study aimed to evaluate the efficacy of a ready-to-use PCV2d and Mycoplasma hyopneumoniae combined vaccine in piglets with different PCV2 MDA levels at vaccination in an experimental inoculation with a heterologous viral genotype. Methods: Forty-eight piglets were allocated into vaccinated (V) and non-vaccinated (NV) groups with high (H) and low (L) PCV2 MDA subgroups (H-V, H-NV, L-V, L-NV). At 3 weeks of age, the piglets received either one dose of vaccine or placebo. Five weeks later, all animals were intranasally challenged with a PCV2b inoculum. Body weight was registered at different time points. Blood samples, peripheral blood mononuclear cells and tracheobronchial lymph nodes (TBLN) were collected and used to assess viraemia, viral load, humoral and cellular responses and histological lesions. Results: The V group showed higher PCV2 antibody levels from challenge onwards, along with a lower percentage of viraemic pigs and reduced viral load in serum at 2 and 3 weeks post-challenge (wpc) and in TBLN tissues compared to the NV group. The H-V group had the highest antibody levels post-challenge, showed no detectable viraemia and had a lower overall amount of virus in tissues. The NV group (especially H-NV) exhibited increased levels of IFN-γ, IFN-α and TNF-α post-challenge. Conclusions: The tested vaccine elicited humoral and cellular immune responses and reduced viral presence in serum and tissues, demonstrating efficacy in a PCV2 subclinical infection model despite high MDA levels at the time of vaccination. Understanding both humoral and cellular immune responses according to different MDA levels can help design more effective vaccination strategies against PCV2. Full article

Durum wheat is a vital wheat species cultivated worldwide for human consumption, ranking second to bread wheat. The Ethiopian durum wheat allele pool shows wide gene diversity; however, limited improvement work has been done to exploit this diversity. Thus, this study aimed to assess the genetic variability, heritability, and interrelationship among different phenotypic traits in 210 recombinant inbred lines (RILs) using an alpha lattice design with two replications. The analysis of variance revealed a significant difference for all the measured traits. The phenotypic coefficient of variation (PCV) was greater than the genotypic coefficient of variation (GCV) for all the characters, which reflects that the existing range of variability within the genotypes was not only due to the varying influence of genotype but also the environment. A correlation analysis disclosed that grain yield was positively related to the traits of plant height and 1000-kernel weight, suggesting that selecting these traits could enhance yield. Path analysis revealed that days to booting, maturity, and 1000-kernel weight directly affect grain yield. Among the measured traits, early developmental traits revealed higher broad-sense heritability. The findings of this study highlight high genetic diversity among Ethiopian durum wheat genotypes, opening up opportunities to integrate these materials into future wheat-breeding programs through introgression with other germplasm sources in Ethiopia and beyond. Full article

The occurrence of PCV2 genotypes in domestic pig production is a dynamic process that undergoes continuous change. Beginning with PCV2a as the first recognized genotype, PCV2b, and subsequently PCV2d, has become the most prevalent one over time. The present study provides an update on the prevalence of the three major PCV2 genotypes in Germany in 2024. A total of 87 fattening farms were randomly selected, proportionally based on farm density within the respective federal states. On each farm, oral fluid samples (OFs) were collected from approximately 100 pigs aged 18 (±1) weeks. Oral fluids (OFs) were pooled and screened for PCV2 DNA by qPCR. Positive samples were subsequently examined by genotype specific qPCR. In total, 31.0% (27/87) of all farms were identified as PCV2-positive. PCV2a was detected in 8.0% (7/87) of farms, while 3.4% (3/87) tested positive for both PCV2a and PCV2d. Overall, 11.5% (10/87) of all farms were PCV2d-positive. No significant effect of vaccination status of the pigs on the viral load or frequency of detection of PCV2 DNA was detected. Full article

Porcine circoviruses are significant pathogens that affect swine populations worldwide, with implications for animal health and productivity. While PCV2 is well-documented, particularly due to widespread vaccination programs, PCV3 is less understood, and its epidemiological impact is still under investigation. This study screened for PCV2 and PCV3 in pigs and wild boars across Portugal to assess their prevalence. Also, nucleotide sequence determination was performed to evaluate the genetic diversity of these viruses. Stool samples from 160 pigs belonging to different groups (quarantine, nursery, fattening and adult pigs), as well as organ samples from 120 hunted wild boars, were analyzed. Samples were collected from twelve of the eighteen mainland Portuguese districts with positive cases being detected in nine of them. Pigs had a lower prevalence of PCV2 (1.9%) than PCV3 (11.2%), but the opposite was true in wild boars (76.7% for PCV2 and 55.0% for PCV3). The lower PCV2 prevalence in pigs can be attributed to the PCV2 vaccination program implemented. Additionally, these viruses were significantly more prevalent in wild boars (90.8% were infected with at least one of the viruses) than in domestic pigs (only 12.5%). This significant difference highlights the impact of the controlled environment in pig farms on disease prevention in contrast to the higher exposure risks faced by wild boars in their natural habitat. Compared to a previous study from 2023, we observed a slight decrease in the percentage of positive cases for both PCV2 and PCV3. Phylogenetic analysis of sequences obtained by Sanger sequencing allowed us to conclude that the samples from domestic pigs belong to the PCV2a and PCV3c clades, in contrast to the PCV2-positive cases detected in domestic pigs in 2023 that were classified in the PCV2d genotype. Conversely, samples from wild boars belong to the PCV2d and PCV3a clades. These results reveal genotype differences between wild and domestic pigs and shifts from 2023 to 2024. Our findings provide some information about the circulation of these viruses and emphasize the importance of vaccination and continued monitoring for a deeper understanding of their epidemiology to mitigate potential risks to swine health and production. Full article

Background/Objectives: Porcine circovirus type 2d (PCV2d) is the predominant genotype associated with porcine circovirus-associated disease (PCVAD), leading to significant economic losses. In South Korea, current vaccine lot-release testing relies on a T/C-ratio-based guinea pig assay, which lacks scientific justification and methodological robustness. This study aimed to develop and validate a statistically defined in-house ELISA using rabbit-derived polyclonal antibodies against PCV2d for the standardized evaluation of immunogenicity. Methods: Polyclonal IgG was generated by immunizing a rabbit with inactivated PCV2d, and it was purified through Protein A chromatography. Guinea pigs (n = 18) were immunized with IMMUNIS^® DMVac, an inactivated PCV2d vaccine candidate developed by WOOGENE B&G, at different doses. In-house ELISA parameters were optimized (antigen coating, blocking agent, and substrate incubation), and analytical performance was evaluated by ROC, linearity, reproducibility, and specificity. Sera from guinea pigs and pigs were analyzed under validated conditions. Results: The optimal performance was achieved using 10⁵ genomic copies/mL of the antigen coating and a 5% BSA blocking agent. The assay showed strong diagnostic accuracy (AUC = 0.97), reproducibility (CVs < 5%), and linearity (R² = 0.9890). Specificity tests with PCV2a, PCV2b, and PRRSV showed minimal cross-reactivity (<7%). The cross-species comparison revealed a positive correlation (R² = 0.1815) and acceptable agreement (bias = −0.21) between guinea pig and porcine sera. The validated cut-off (S/P = 0.4) enabled accurate classification across both species and aligned well with commercial kits. Conclusions: The in-house ELISA offers a robust, reproducible, and scientifically validated platform for immunogenicity verification, supporting its application in Korea’s national lot-release system. Homologous competition assays with PCV2d are planned to further confirm antigen specificity. Full article

Background: Porcine circovirus disease (PCVD), caused by porcine circovirus (PCV), is a significant swine disease characterized by porcine dermatitis, nephrotic syndrome, and reproductive disorders in sows. Given the overlapping clinical presentations of PCV2, PCV3, and PCV4, a rapid and accurate method for their differential detection is essential. Methods: In this study, specific primers and probes were designed based on the conserved regions of the ORF1 genes of PCV2 and PCV4, as well as the ORF2 gene of PCV3. Results: A TaqMan triple real-time PCR method was developed, demonstrating excellent specificity, sensitivity, and repeatability, with limits of detection (LODs) of 53.3 copies/µL, 12.0 copies/µL, and 13.8 copies/µL for PCV2, PCV3, and PCV4, respectively. Using this method, 500 clinical porcine tissue samples collected from 23 provinces across China between 2023 and 2024 were analyzed. The results showed detection rates of 75.20% (376/500) for PCV2, 17.60% (88/500) for PCV3, and 4.40% (22/500) for PCV4. The detection rate of triple coinfections involving PCV2, PCV3, and PCV4 was 0.80% (4/500). PCV2 consistently presented significantly higher positive detection rates across all growth stages, and its viral copy number was significantly greater than those of PCV3 and PCV4 (* p < 0.05). Forty PCV2 ORF2 genes, fourteen PCV3 ORF2 genes, and three PCV4 ORF2 genes were identified. These included four PCV2a genotypes, thirty-five PCV2d genotypes, and one PCV2e genotypes; two PCV3a genotypes and six each of PCV3b and PCV3c genotypes; and two PCV4a genotypes and one of PCV4b genotype. Conclusions: The triple qPCR method established in this study provides a rapid, specific, and accurate approach for the detection and differentiation of PCV2, PCV3, and PCV4 genotypes. Full article

Mycoplasma hyopneumoniae (Mhyo) and porcine circovirus type 2 (PCV2) are critical pathogens in the swine industry, both contributing significantly to the porcine respiratory disease complex (PRDC). Given their impact, it is logical to control these pathogens simultaneously. Consequently, combined vaccinations against Mhyo and PCV2 are gaining popularity in swine health management. We present the efficacy of the first commercial combined vaccine prepared of a genotype PCV2d strain and Mhyo and tested against experimental challenge infections with target pathogens in comparative trials with other commercial products. In these studies, three-week-old piglets were vaccinated according to the manufacturers’ instructions. Five weeks later, they were challenged with two Mhyo strains over three consecutive days or with a PCV2d strain once. Positive controls included challenged pigs without prior vaccination, while non-vaccinated/non-challenged pigs served as negative controls. The key parameters measured were lung lesion scores and seroconversion for Mhyo, and viraemia, rectal shedding, lymph node and lung viral content, and seroconversion for PCV2. Findings and conclusion: The results showed no compromising effects between the vaccine components and highlighted significant differences in efficacy among the various products tested. Additionally, oral fluid sampling demonstrated a strong correlation with the viraemia and fecal shedding of PCV2, underscoring the diagnostic and animal welfare benefits of this sampling method. Full article

The widespread distribution and genetic diversity of porcine circovirus type 2 (PCV2) seriously threatens the swine industry worldwide. This study investigates the molecular epidemiology of PCV2 in Henan Province (2020–2023) through PCR screening (385 samples) and whole-genome sequencing (34 strains). The overall detection rate was 71.17% (274/385), with annual rates of 81.16% (112/138) in 2020, 72.41% (84/116) in 2021, 62.50% (55/88) in 2022, and 53.49% (23/43) in 2023, indicating a declining trend. Phylogenetic analysis revealed the dominance of the PCV2d genotype, comprising 82.4% (28/34) of sequenced strains. Evolutionary analysis identified strong negative selection pressure on ORF2, with an elevated substitution rate of 1.098 × 10⁻³ ssy. These findings provide critical insights into the predominance and adaptive evolution of PCV2d, and significantly improve our understanding of its genetic diversity and evolutionary dynamics. Full article

African Swine Fever Virus (ASFV) genotype II dominates outbreaks in Tanzania’s Southern Highlands, continuing to persist as the dominant strain over a decade after its first incursion in 2010. A total of 205 samples from 120 holdings were collected, with 21 confirmed ASFV-positive animals from 14 holdings. Molecular analysis revealed genetic uniformity among isolates, all clustering within ASFV genotype II. Poor biosecurity measures, such as feeding of untreated swill (80% of holdings) and lack of restrictions on visitors (90% of holdings), were identified as risk factors. Additionally, co-infection with porcine circovirus-2 (PCV-2) further complicates disease management. This study underscores the urgent need for enhanced biosecurity and farmer education to mitigate ASFV outbreaks in endemic regions. Full article

Pigs are susceptible to the deadly infectious disease known as African swine fever (ASF), which is brought on by the African swine fever virus (ASFV). As such, prompt and precise disease detection is essential. Deletion of the virulence-related genes MGF-505/360 and EP402R generated from the virulent genotype II virus significantly reduces its virulence, and animal tests using one of the recombinant viruses show great lethality and transmissibility in pigs. The isothermal technique known as recombinase polymerase amplification (RPA) is perfect for rapid in-field detection. To accurately identify ASFV MGF-505R gene-deleted mutants and assess the complex infection situation of ASF, RPA assays in conjunction with real-time fluorescent detection (real-time RPA assay) and lateral flow dipstick (RPA-LFD assay) were created. These innovative methods allow for the direct detection of ASFV from pigs, offering in-field pathogen detection, timely disease management, and satisfying animal quarantine requirements. The specific primers and probes were designed against conserved regions of ASFV B646L and MGF-505R genes. Using recombinant plasmid DNA containing ASFV MGF-505R gene-deleted mutants as a template, the sensitivity of both ASF real-time RPA and ASF RPA-LFD assays were demonstrated to be 10 copies per reaction within 20 min at 37 °C. Neither assay had cross-reactions with CSFV, PRRSV, PPV, PRV, ot PCV2, common viruses seen in pigs, indicating that these methods were highly specific for ASFV. The evaluation of the performance of ASFV real-time RPA and ASFV RPA-LFD assays with clinical samples (n = 453) demonstrated their ability to specifically detect ASFV or MGF-505R gene-deleted mutants in samples of pig feces, ham, fresh pork, and blood. Both assays exhibited the same diagnostic rate as the WOAH-recommended real-time fluorescence PCR, highlighting their reliability and validity. These assays offer a simple, cost-effective, rapid, and sensitive method for on-site identification of ASFV MGF-505R gene-deleted mutants. As a promising alternative to real-time PCR, they have the potential to significantly enhance the prevention and control of ASF in field settings. Full article