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Pseudorabies virus (PRV), causing Aujeszky's disease in swine, has important economic impact on the pig industry in China and even poses a threat to public health. Although this disease has been controlled by vaccination with PRV live attenuated vaccines (LAVs), the potency of PRV LAVs in inducing cellular immunity has not been well characterized. In this study, using PRV Bartha K61 strain (BK61), the most-used PRV LAVs, as a model, we re-examined the cellular immune response elicited by the BK61 in mice and pigs by multicolor flow cytometry. We found that phenotypic activation of T cells, NK cells and B cells was hardly detected after vaccination. However, antigen-specific IFN-γ-producing CD4 T cells rather than CD8 T cells were dominantly detected but at low frequency upon restimulation with live BK61 virus. These BK61-specific CD4 T cells are also able to simultaneously produce TNF-α and IL-2, showing characteristics of multifunctional T cells. However, BK61-specific CD4 T cells showed weak secondary response upon challenge with PRV DX strain. Further vaccination with PRV-infected dendritic cells (DCs) transiently increased the percentage of IFN-γ-positive CD4 and CD8 T cells but eventually restored to low frequency and did not improve the protective efficacy of BK61 against challenge, suggesting that PRV BK61 induced a relatively weak cellular immunity that could not be overcome by the DC vaccination. Similar immune responses were also observed following vaccination with another PRV LAV HD/c in mice and pigs, suggesting that this may be an intrinsic drawback of PRV LAVs in inducing cellular immunity. Our results demonstrated that PRV LAVs elicited a CD4 Th1-biased weak cellular immunity which is implicative for the development of PRV-vectored vaccine. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS) is an endemic disease affecting the swine industry. The disease is caused by the PRRS virus (PRRSV). Despite extensive biosecurity and control measures, the persistence and seasonality of the virus have raised questions about the virus's environmental dynamics during the fall season when the yearly epidemic onset begins and when crop harvesting and manure incorporation into the field occur. Therefore, this study aimed to assess the potential for PRRSV to percolate through different soil types, simulating conditions that could lead to groundwater contamination which could represent a risk of herd introduction. An experimental soil column model was used to mimic field conditions. Three PRRSV-2 strains were tested across thirteen Minnesota soils with different physical and chemical characteristics. The findings revealed that PRRSV can percolate through all soil types and that the amount of virus percolated decreases with increased amounts of soil. These results suggest that PRRSV can percolate through different soil types. Further investigations should be undertaken to determine the associated implications for swine health and biosecurity measures. [ABSTRACT FROM AUTHOR]

Coronaviruses continue to disrupt health and economic productivity worldwide. Porcine epidemic diarrhea virus (PEDV) is a devastating swine disease and SARS-CoV-2 is the latest coronavirus to infect the human population. Both viruses display a similar spike protein on the surface that is a target of vaccine development. Despite the availability of commercial vaccines for both viruses, there is still a high occurrence of infections and a great need for enhanced efficacy and lower costs. We previously produced the PEDV spike protein (S) using transgenic maize, enabling a low-cost supply of the vaccine candidate. In this study, we (1) test orally delivered PEDV vaccine candidates in pigs to optimize the mucosal immune response; (2) generate the SARS-CoV-2 S1 protein in maize; and (3) perform structural characterization of the S1 protein for PEDV and SARS-CoV-2. We demonstrated high expression levels in maize of the S1 subunit of the SARS-CoV-2 spike protein, both with and without fusion to the heat-labile enterotoxin B (LTB) subunit. We found that the LTB fusion protein from both coronaviruses preferentially assembles into higher molecular weight multimers, consistent with the formation of trimers. For PEDV, administering the spike protein fused to LTB to young pigs elicited a higher level of mucosal IgAs compared to maize grain containing the S1 protein alone or controls. This suggests that fusing the coronavirus spike protein with LTB may provide better protection. [ABSTRACT FROM AUTHOR]

Viral infections in swine, such as African swine fever (ASF), porcine reproductive and respiratory syndrome (PRRS), and foot-and-mouth disease (FMD), have a significant impact on the swine industry. Despite the significant progress in the recent efforts to develop effective vaccines against viral diseases in swine, the search for new protective vaccination strategy remains a challenge. The antigenic epitope, acting as a fundamental unit, can initiate either a cellular or humoral immune response. Consequently, the combination of multi-epitopes expressing different stages of viral life cycle has become an optimal strategy for acquiring a potent, safe, and effective vaccine for preventing and treating viral diseases in swine. Recent progresses in immunoinformatic tools, coupled with an understanding of host immune responses and computational biology, have paved the way for innovative vaccine design disciplines that focus on computer-assisted, in-silico epitope prediction for the prevention of viral diseases in swine. The concept of multi-epitope vaccines driven by immunoinformatic methods has gained prominence in multiple studies, particularly in the development of vaccines targeting conserved epitopes in variable or rapidly mutating pathogens such as African swine fever virus (ASFV) and porcine reproductive and respiratory syndrome virus (PRRSV). In this review, we provide an overview of the in-silico design of the multi-epitope vaccines against viral diseases in swine, including the antigenicity, structural quality analysis, immune simulations, and molecular dynamics (MD) simulations. Furthermore, we also enumerate several multi-epitope vaccine applications that have shown promise to be against viral diseases in swine. [ABSTRACT FROM AUTHOR]

PCV2 is a significant epidemic agricultural pathogen that causes a variety of swine diseases. PCV2 infections have significant economic impact on the swine industry, making effective strategies for rapid detection of PCV2 in pigs essential. Herein, we report on the synthesis of the so-called nano-MIPs which can be utilized for molecular recognition of PCV2. The morphology and structure of nano-MIPs were characterized using scanning electron microscopy (SEM). Nano-MIPs are spherical with sizes around 120–150 nm. Binding experiments demonstrate that the fluorescence intensity of PCV2 samples decreases proportionally to increasing the concentration of nano-MIPs due to quenching, while non-imprinted polymer nanoparticles (nano-NIPs) do not affect the signal. The Stern–Volmer constant of nano-MIPs binding to PCV2 was 1.3 × 10−3 mL/µg, whereas nano-NIPs led to 7 × 10−5 mL/µg, i.e., 1.8 orders of magnitude lower. The detection limit for binding MIP particles to PCV2 by fluorescence measurements is 47 µg/mL. This affinity test allows for designing both direct and competitive quartz crystal microbalance (QCM) assays for PCV2 leading to QCM measurements. The QCM results show nano-MIPs binding to PCV2 immobilized on the sensor surface with appreciable reproducibility. QCM sensor characteristics reveal signal saturation above around 200 µg/mL at a response of − 354 Hz and an LOD of approximately 35 µg/mL. Nano-MIPs also show selectivity factors of 2–5 for CSFV and PRRSV probably because the three viruses have similar diameters around 50 nm. [ABSTRACT FROM AUTHOR]

Background: Respiratory disease is an economically important disease in the swine industry. Housing air quality control is crucial for maintaining the respiratory health of pigs. However, maintaining air quality is a limitation of current housing systems. This study evaluated the growth and health parameters of pigs raised under different environmental conditions and identified key environmental variables that determine respiratory health. Eighty (Largewhite × Landrace) × Duroc crossed growing pigs (31.71 ± 0.53 kg) were equally distributed into two identical climate-controlled houses with distinct environmental conditions (CON = normal conditions and TRT = poor conditions). Two-sample tests were performed to compare the means of the groups, and a random forest algorithm was used to identify the importance scores of the environmental variables to respiratory health. Results: Pigs in the TRT group were significantly exposed to high temperatures (28.44 vs 22.78 °C, p < 0.001), humidity (88.27 vs 61.86%, p < 0.001), CO2 (2,739.93 vs 847.91 ppm, p < 0.001), NH3 (20.53 vs 8.18 ppm, p < 0.001), and H2S (14.28 vs 6.70 ppm, p < 0.001). Chronic exposure to these factors significantly reduced daily feed intake (1.82 vs 2.32 kg, p = 0.002), resulting in a significant reduction in average daily gain (0.72 vs 0.92 kg, p = 0.026), increased oxidative stress index (3.24 vs 1.43, p = 0.001), reduced cortisol levels (2.23 vs 4.07 mmol/L, p = 0.034), and deteriorated respiratory health status (74.41 vs 97.55, p < 0.001). Furthermore, a random forest model identified Min CO2, Min NH3, and Avg CO2 as the best predictors of respiratory health, and CO2 was strongly correlated with NH3 and H2S concentrations. Conclusions: These findings emphasize the critical importance of proper environmental management in pig farming and suggest that regular monitoring and control of either CO2 or NH3, facilitated by environmental sensors and integration into intelligent systems, can serve as an effective strategy for improving respiratory health management in pigs. [ABSTRACT FROM AUTHOR]

Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia, a challenging respiratory disease for the global swine industry. Variations in the serotypes associated with clinical disease have been observed in different regions worldwide. This study aimed to provide an updated epidemiological assessment of A. pleuropneumoniae serotypes in Spain, incorporating bacterial characterization through serotyping and toxinotyping. Serotypes 9/11, 2, 4, 5, 17, and 13 were frequently identified in diseased animals. Furthermore, qPCR of lung samples from an outbreak, even when samples were pooled, emerged as a robust diagnostic tool, enabling the rapid detection of A. pleuropneumoniae and their serotypes without the need for microbiological isolation. This technology also facilitates serotype monitoring of apparently healthy herds through the testing of oral fluids. The study revealed the frequent simultaneous presence of diverse serotypes within a farm. Serotypes 1, 7, 10, 12, 18, and 19 were frequently found in subclinically infected animals but were rarely detected in acute pleuropneumonia outbreaks in the current study. These results provide valuable information for interpreting the potential virulence of the different serotypes in Spain. However, other predisposing factors and the immune status of the herds such as type of vaccines used when appropriate, should be carefully considered before drawing definitive conclusions. Nevertheless, the study offers valuable insights that underscore the necessity for detailed regional data to contribute toward a comprehensive understanding of the disease dynamics and toward formulating effective control measures for porcine pleuropneumonia. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) is a fatal infectious disease of swine caused by the African swine fever virus (ASFV). Currently, the disease is listed as a legally notifiable disease that must be reported to the World Organization for Animal Health (WOAH). The economic losses to the global pig industry have been insurmountable since the outbreak of ASF. Control and eradication of ASF are very critical during the current pandemic. Vaccination is the optimal strategy to prevent and control the ASF epidemic, but since inactivated ASFV vaccines have poor immune protection and there aren't enough cell lines for efficient in vitro ASFV replication, an ASF vaccine with high immunoprotective potential still remains to be explored. Knowledge of the course of disease evolution, the way of virus transmission, and the breakthrough point of vaccine design will facilitate the development of an ASF vaccine. In this review, the paper aims to highlight the recent advances and breakthroughs in the epidemic and transmission of ASF, virus mutation, and the development of vaccines in recent years, focusing on future directions and trends. [ABSTRACT FROM AUTHOR]

Host and pathogen dynamics are driven by seasonal variation, leading to complex epidemic patterns which are not well understood. Here, we linked demography and epidemiology in a single framework to reach a mechanistic understanding of disease dynamics in natural populations characterized by seasonality. We applied this approach to depict the interplay between a host, the wild boar, and a viral disease, African swine fever. This virus spreads in wild boar by direct‐contact among living individuals or though contacts with infected carcasses. In this system, both host and pathogen show seasonal patterns: survival and reproduction probabilities vary across the year in the wild boar, and virus persistence in the environment is affected by temperature which influences the speed of carcasses degradation. Using a stochastic individual‐based model, we showed that virus invasion success strongly varies according to the time of introduction due to the seasonal specific interaction between the virus and host. Furthermore, the shape of the following epidemic varies across seasons, with variation in speed and magnitude of the outbreak. Consequently, the host population response was also season‐specific, both regarding the speed of the decline and the change in population structure. Overall, our study highlights the inextricable connections between host, pathogen and the environmental dynamics, and the importance to consider each component to understand disease dynamics. [ABSTRACT FROM AUTHOR]

A priority to facilitate the application of lipofection to generate genetically modified porcine embryos and animals will be the use of zona pellucida (ZP)-intact oocytes and zygotes. Recently, our group produced genetically modified embryos by lipofection of ZP-intact oocytes during in vitro fertilization (IVF). This study investigates the effect of two commercial lipofection reagents, Lipofectamine 3000 and Lipofectamine CRISPRMAX, on embryo development and mutation efficiency in ZP-intact porcine oocytes. We compared these reagents with the electroporation method and a control group using two sgRNAs targeting the CAPN3 and CD163 genes. The detrimental effects on cleavage rates were observed in both lipofection treatments compared to the control and electroporated groups. However, blastocyst rates were higher in the Lipofectamine 3000 group than in the electroporated group for both genes. Mutation parameters varied by target gene, with Lipofectamine 3000 achieving higher mutation rates for CD163, while all groups were similar for the CAPN3 gene. Overall efficiency was similar for both lipofectamines, confirming their feasibility for use. In addition, we evaluated the effect of coincubation time (4, 8, and 24 h) on IVF outcomes, embryo development, and mutation parameters. Results indicated that an 8-h coincubation period optimized fertilization and mutation efficiency without significant toxic effects. This study demonstrates that lipofection with either Lipofectamine 3000 or CRISPRMAX during IVF is an effective method for generating genetically modified porcine embryos without the need for specialized equipment or trained personnel, with efficiencies similar to or greater than electroporation. This study also highlights the importance of optimizing reagent selection and coincubation times. There is no difference between Lipofectamine 3000 and CRISPRMAXTM in terms of embryo development and mutation efficiency, and under our experimental conditions, the optimal coincubation time with lipofectamine is 8 h. • Lipofection during IVF is an effective method to generate mutant porcine embryos. • Lipofectamine 3000 and CRISPRMAX are similar in efficiency. • Lipofection maintains high blastocyst rates. • Optimal coincubation time with Lipofectamine is 8 h. [ABSTRACT FROM AUTHOR]

Pen-based oral fluids are used extensively for surveillance and disease detection in swine, but there is sparse information on the sampling process itself. To address this shortcoming, we documented the pen-based oral fluid sampling process with the aim of optimizing the number of pigs in a pen that contributed to the sample. We quantified the effects of (1) previous experience with rope sampling (training), (2) the number of ropes suspended in the pen, and (3) sampling time on pig participation and pig-rope contact. A subset of pigs was clearly marked for individual identification and their interactions with ropes video recorded. Thereafter, pig-rope contacts were counted from the recordings, with "contact" defined as an individually identified pig clearly taking the rope into its mouth. Data were analyzed using appropriate models (R version 4.4.1 R core team 2024). Training, provision of additional ropes, and extended sampling time all increased pig participation across pen sizes. However, for routine oral fluid collection in the field, we recommend training pigs prior to hanging ropes for sample collection and increasing sampling time to maximize the pigs' contribution to the oral fluid sample. Importantly, these studies focused on pig behavior and not detection; thus, future studies should evaluate the impact of these same factors on the probability of detection. [ABSTRACT FROM AUTHOR]

Porcine astrovirus 4 (PoAstV4) has been recently associated with respiratory disease in pigs. In order to understand the scope of PoAstV4 infections and to support the development of a vaccine to combat PoAstV4 disease in pigs, we designed and produced a recombinant PoAstV4 capsid spike protein for use as an antigen in serological assays and for potential future use as a vaccine antigen. Structural prediction of the full-length PoAstV4 capsid protein guided the design of the recombinant PoAstV4 capsid spike domain expression plasmid. The recombinant PoAstV4 capsid spike was expressed in Escherichia coli, purified by affinity and size-exclusion chromatography, and its crystal structure was determined at 1.85 Å resolution, enabling structural comparisons to other animal and human astrovirus capsid spike structures. The recombinant PoAstV4 capsid spike protein was also used as an antigen for the successful development of a serological assay to detect PoAstV4 antibodies, demonstrating that the recombinant PoAstV4 capsid spike retains antigenic epitopes found on the native PoAstV4 capsid. These studies lay a foundation for seroprevalence studies and the development of a PoAstV4 vaccine for swine.

African swine fever virus (ASFV), which poses significant risks to the global economy, encodes a unique host-independent transcription system. This system comprises an eight-subunit RNA polymerase (vRNAP), temporally expressed transcription factors and transcript associated proteins, facilitating cross-species transmission via intermediate host. The protein composition of the virion and the presence of transcription factors in virus genome suggest existence of distinct transcription systems during viral infection. However, the precise mechanisms of transcription regulation remain elusive. Through analyses of dynamic transcriptome, vRNAP-associated components and cell-based assay, the critical role of M1249L in viral transcription regulation has been highlighted. Atomic-resolution structures of vRNAP-M1249L supercomplex, exhibiting a variety of conformations, have uncovered the dual functions of M1249L. During early transcription, M1249L could serve as multiple temporary transcription factors with C-terminal domain acting as a switcher for activation/inactivation, while during late transcription it aids in the packaging of the transcription machinery. The structural and functional characteristics of M1249L underscore its vital roles in ASFV transcription, packaging, and capsid assembly, presenting novel opportunities for therapeutic intervention. African swine fever virus (ASFV) is a contagious DNA virus causing lethal disease in swine. This study reveals structures of ASFV RNA polymerase bound to protein M1249L, showing its vital roles in regulating early transcription and packaging. [ABSTRACT FROM AUTHOR]

Pradofloxacin is a dual targeting, bactericidal fluoroquinolone recently approved for treating bacteria causing swine respiratory disease. Currently, an abundance of in vitro data does not exist for pradofloxacin. We determined the minimum inhibitory concentration (MIC) and mutant prevention concentrations (MPC) of pradofloxacin compared to ceftiofur, enrofloxacin, florfenicol, marbofloxacin, tildipirosin, tilmicosin and tulathromycin against swine isolates of Actinobacillus pleuropneumoniae and Pasteurella multocida. Overall, pradofloxacin had the lowest MIC and MPC values as compared to the other agents tested. For example, pradofloxacin MIC values for 50%, 90% and 100% of A. pleuropneumoniae strains were ≤0.016 µg/mL, ≤0.016 µg/mL and ≤0.016 µg/mL and for P. multocida were ≤0.016 µg/mL, ≤0.016 µg/mL and 0.031 µg/mL, respectively. The MPC values for 50%, 90% and 100% of A. pleuropneumoniae strains were 0.031 µg/mL, 0.063 µg/mL and 0.125 µg/mL and for P. multocida were ≤0.016 µg/mL, 0.031 µg/mL and 0.0.063 µg/mL, respectively. By MPC testing, all strains were at or below the susceptibility breakpoint. Based on MPC testing, pradofloxacin appears to have a low likelihood for resistance selection. This study represents the most comprehensive in vitro comparison of the above noted drugs and the first report for pradofloxacin and tildipirosin. [ABSTRACT FROM AUTHOR]

Background: African swine fever (ASF) is one of the deadliest swine diseases with haemorrhagic symptoms and a high mortality rate. Plant‐derived additives are potential antiviral agents against viruses due to their environmental and user‐friendly properties. Objectives: This study aims to evaluate the efficacy of plant‐based additives (Phyto.A04 and Phyto.B) compared to an organic acid blend (OAB) in inactivating ASF virus (ASFV) in cell culture and feed. Methods: ASFV‐spiked feed was treated with individual or combined additives such as OAB, Phyto.A04 and Phyto.B. The viability of ASFV after treatment of ASFV‐spiked feed with additives was then confirmed by both methods, real‐time PCR and cell culture. Results: The results of the in vitro test with cell cultures showed that all three additives (OAB, Phyto.A04 and Phyto.B) exerted a strong virucidal effect on ASFV in porcine alveolar macrophage cells. OAB at a concentration of 0.3% reduced the virus concentration from 4.48 log10 HAD50/mL after 1 day of treatment (day 1) to 3.29 log10 HAD50/mL after 3 days of treatment (day 3) and remained undetected after 7 days of treatment (day 7). In Phyto.A04 with 1%, the virus was only detectable on day 1 (3.53 log10 HAD50/mL). Phyto.B with 0.01% and 0.05% both showed good efficacy in completely inhibiting virus presence on days 3 and 7. Conclusions: All additives, OAB, Phyto.A04 and Phyto.B, were able to inactivate ASFV in a dose‐dependent manner, as confirmed by cell culture and PCR methods. The combination of additives at different concentrations consistently improved the virucidal results. [ABSTRACT FROM AUTHOR]

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Simple Summary: Foot-and-mouth disease (FMD), caused by the foot-and-mouth disease virus (FMDV), is a prevalent infectious disease in swine. Timely detection and isolation of pigs infected with this virus are among the most effective strategies to prevent the disease's spread and mitigate potential economic damage. We have established a set of RT-qPCR detection methods for FMDV, which offer significant advantages. Notably, this detection method can effectively identify foot-and-mouth disease virus even in the presence of both exogenous and endogenous interfering substances. Furthermore, it exhibits high sensitivity, strong specificity, and good repeatability, enabling 100% detection of FMDV. The global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its sensitivity, specificity, reproducibility, and stability. The standard curve equation range is 6.43 × 109–6.43 × 101 copies/µL, with an R2 value of 0.996 and a standard curve equation of y = −3.586x + 36.245. The method successfully detected 64.3 copies/µL of the target gene for FMDV and exhibited high specificity for FMDV. Repeatability tests demonstrated low coefficients of variation within and between groups (<2%), indicating good reproducibility. The clinical samples analyzed using this method showed results consistent with those of the SYBR Green I RT-qPCR assay, confirming the reliability of the method. Overall, the developed test method displayed high sensitivity, specificity, reproducibility, and stability, making it suitable for the rapid diagnosis of foot-and-mouth disease in clinical settings. [ABSTRACT FROM AUTHOR]

Classical swine fever (CSF), sometimes referred to as hog cholera, is a highly contagious, virally based, systemic illness that affects both domestic and wild pigs. The virus known as classical swine fever virus (CSFV) is a member of the Flaviviridae family, specifically the genus Pestivirus. This disease is thought to be endemic in many Asian countries that produce pork as well as in several countries in Central and South America, the Caribbean, and elsewhere. As previously indicated, depending on the virulence of the virus strain involved and several host circumstances, clinical indications of CSFV infection can vary greatly, ranging from abrupt fatality to an occult course. CSF diagnosis can be made by serological detection, antigen, RNA, and isolation. CSF's highly varied symptoms and post-mortem pathology resemble those of African swine fever (ASF). ASF, the kind of CSFV, the pig's age, and its susceptibility all affect the clinical symptoms. Pigs that contract CSFV, a highly infectious and economically significant virus. The great economic significance of the swine business makes the CSFV a potential bioterrorism threat. Live attenuated CSF vaccinations have been around for many years and are quite safe and effective. Controlling epidemics in CSFV-free zones requires quick action. Pigs that are impacted must be slaughtered, and the carcasses must be buried or burned. [ABSTRACT FROM AUTHOR]

Simple Summary: Spatial epidemiology, integrating traditional epidemiology, geography, statistics, environmental science, and ecology, provides a comprehensive framework for analyzing the spatial dimensions of health and disease. This interdisciplinary approach enhances the development of effective public health strategies and interventions. However, its multifaceted nature can bring complexities in practical application. Using the case of spatial epidemiology in swine viral diseases (SVDs), we illustrate the objectives, methodologies, and essential considerations for the application of spatial epidemiology, which we hope to offer as a comprehensive reference for researchers in this field. Spatial epidemiology offers a comprehensive framework for analyzing the spatial distribution and transmission of diseases, leveraging advanced technical tools and software, including Geographic Information Systems (GISs), remote sensing technology, statistical and mathematical software, and spatial analysis tools. Despite its increasing application to swine viral diseases (SVDs), certain challenges arise from its interdisciplinary nature. To support novices, frontline veterinarians, and public health policymakers in navigating its complexities, we provide a comprehensive overview of the common applications of spatial epidemiology in SVD. These applications are classified into four categories based on their objectives: visualizing and elucidating spatiotemporal distribution patterns, identifying risk factors, risk mapping, and tracing the spatiotemporal evolution of pathogens. We further elucidate the technical methods, software, and considerations necessary to accomplish these objectives. Additionally, we address critical issues such as the ecological fallacy and hypothesis generation in geographic correlation analysis. Finally, we explore the future prospects of spatial epidemiology in SVD within the One Health framework, offering a valuable reference for researchers engaged in the spatial analysis of SVD and other epidemics. [ABSTRACT FROM AUTHOR]

The objective of the study was to identify risk factors for reintroduction of Actinobacillus pleuopneumoniae and Mycoplasma hyopneumoniae (enzootic pneumonia) onto pig farms in areas in Switzerland that were involved in an eradication programme from 1996 to 1999 and to assess the role of dealers in relation to these reinfections. The study was based on the comparison of pig farms that were reinfected in the year 2000 (cases) and pig farms that remained uninfected in the same area (controls). Additionally, data were collected from Swiss pig dealers and transport companies. Out of a total of 3983 farms, 107 farms were reinfected in the year 2000. The incidences were 0.1% for Actinobacillus pleuopneumoniae and 2.6% for Mycoplasma hyopneumoniae (enzootic pneumonia). Compared to reinfection rates prior to the eradication programme, this is a considerable reduction. Statistically significant risk factors for the reinfection were 'finishing farm', 'large mixed breeding-finishing farm', 'reinfected neighbour' and 'parking site for pig transport vehicles close to the farm'. Pig farmers that purchased pigs from only one supplier per batch had a lower risk of reintroducing infection (protective factor). As long as infected and uninfected regions co-exist in Switzerland, direct and indirect contact between farms, pig herds and slaughter sites via transport vehicles are a major pathway of disease spread. Risk management measures linked to these contacts are therefore of key importance. The survey of dealers indicated various areas for improvement such as strategic planning of pick-up routes or cleaning and disinfecting of trucks. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) is a highly contagious and lethal disease of swine caused by African swine fever virus (ASFV), and the mortality rate caused by virulent stains can approach 100%. Many ASFV viral proteins suppress the interferon production to evade the host's innate immune responses. However, whether ASFV MGF360-4L could inhibit type I interferon (IFN-I) signaling pathway and the underlying molecular mechanisms remain unknown. Our study, indicated that ASFV MGF360-4L could negatively regulates the cGAS-STING mediated IFN-I signaling pathway. Overexpressing ASFV MGF360-4L could inhibit the cGAS/STING signaling pathway by inhibiting the interferon-b promoter activity, which was induced by cGAS/STING, TBK1, and IRF3-5D, and further reduced the transcriptional levels of ISG15, ISG54, ISG56, STAT1, STAT2, and TYK2. Confocal microscopy and immunoprecipitation revealed that MGF360-4L colocalized and interacted with IRF3, and WB revealed that ASFV MGF360-4L suppressed the phosphorylation of IRF3. 4L-F2 (75-162 aa) and 4L-F3 (146-387 aa) were the crucial immunosuppressive domains and sites. Altogether, our study reveals ASFV MGF360-4L inhibited cGAS-STING mediated IFN-I signaling pathways, which provides insights into an evasion strategy of ASFV involving in host's innate immune responses. [ABSTRACT FROM AUTHOR]

Streptococcus suis is a major swine pathogen and zoonotic agent, causing important economic losses to the porcine industry. Here, we used genomics approaches to characterize 251 S. suis isolates recovered from diseased pigs across Belgium, France, Germany, Hungary, the Netherlands, Spain, and the United Kingdom. We identified 13 serotypes, being serotypes 9 and 2 the most prevalent, and 34 sequence types (STs), including 16 novel STs, although ST16 and ST1 dominated the strain population. Phylogenetic analysis revealed complex genetic relationships, notable geographic clustering, and potential differential capacity for capsular switching among serotype 9 isolates. We found antimicrobial resistance (AMR) genes in 85.3% of the isolates, with high frequencies of genes conferring resistance to tetracyclines and macrolides. Specifically, 49.4% of the isolates harbored the tetO gene, and 64.9% possessed the ermB gene. Additionally, we observed a diverse array of virulence-associated genes (VAGs), including the classical VAGs mrp, epf, and sly, with variable presence across different genotypes. The high genetic diversity among European S. suis isolates highlights the importance of targeted antimicrobial use and flexible vaccine strategies. Rapid strain characterization is crucial for optimizing swine health management, enabling tailored interventions like the development of autovaccines to mitigate S. suis infections. [ABSTRACT FROM AUTHOR]

Simple Summary: Wildlife in human-dominated landscapes often faces a range of disturbances that can alter their natural behaviors. Wild boar (Sus scrofa) populations are increasing across Europe, raising concerns about ecological impacts and the spread of diseases such as African swine fever (ASF). This research focuses on the behavioral adaptations of wild boars in response to specific human disturbances. Utilizing advanced biologging technologies, specifically accelerometer and magnetometer combined with dead reckoning methods, fifteen wild boars in a suburban forest near Prague were monitored over a period from February 2020 to July 2021. This study provides insights into the wild boar's resilience, revealing that while the animals are inclined to flee when near disturbances, they predominantly remain in a resting state otherwise. Their most common reaction was to continue resting. These observations underscore the potential role of disturbance management in controlling the spread of zoonotic diseases such as African swine fever (ASF) within wild boar populations. Currently, African swine fever (ASF), a highly fatal disease has become pervasive, with outbreaks recorded across European countries, leading to preventative measures to restrict wild boar (Sus scrofa L.) movement, and, therefore, keep ASF from spreading. This study aims to detail how specific human activities—defined as "car", "dog", "chainsaw", and "tourism"—affect wild boar behavior, considering the disturbance proximity, and evaluate possible implications for wild boar management in ASF-affected areas. Wild boar behavior was studied using advanced biologging technology. This study tracks and analyzes wild boar movements and behavioral responses to human disturbances. This study utilizes the dead reckoning method to precisely reconstruct the animal movements and evaluate behavioral changes based on proximity to disturbances. The sound of specific human activities was reproduced for telemetered animals from forest roads from different distances. Statistical analyses show that wild boars exhibit increased vigilance and altered movement patterns in response to closer human activity, but only in a small number of cases and with no significantly longer time scale. The relative representation of behaviors after disruption confirmed a high instance of resting behavior (83%). Running was the least observed reaction in only 0.9% of all cases. The remaining reactions were identified as foraging (5.1%), walking (5.0%), standing (2.2%), and other (3.8%). The findings suggest that while human presence and activities do influence wild boar behavior, adherence to movement restrictions and careful management of human activity in ASF-infected areas is not a necessary measure if human movement is limited to forest roads. [ABSTRACT FROM AUTHOR]

The African swine fever virus (ASFV) is a large and complex DNA virus that causes a highly lethal disease in swine, for which no antiviral drugs or vaccines are currently available. Studying viral–host protein–protein interactions advances our understanding of the molecular mechanisms underlying viral replication and pathogenesis and can facilitate the discovery of antiviral therapeutics. In this study, we employed affinity tagging and purification mass spectrometry to characterize the interactome of VPS39, an important cellular factor during the early phase of ASFV replication. The interaction network of VPS39 revealed associations with mitochondrial proteins involved in membrane contact sites formation and cellular respiration. We show that the ASFV proteins CP204L and A137R target VPS39 by interacting with its clathrin heavy-chain functional domain. Furthermore, we elaborate on the potential mechanisms by which VPS39 may contribute to ASFV replication and prioritize interactions for further investigation into mitochondrial protein function in the context of ASFV infection. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) is one of the deadliest swine diseases, causing significant economic losses, threatening food security, and limiting pig production in affected countries. In the absence of an effective ASF vaccine, prevention and control of ASF depend mainly on effective biosecurity measures. In this study, the efficacy of SAFER®, a powdered disinfectant containing clay, an acid complex, and the active ingredient thyme essential oil, was tested against the ASF virus. The results showed that ASFV isolate (VNUA/HY/ASF1/Vietnam/2019) was inactivated by 3.5 and 5 Log10HAD50/ml after 20 and 120  min of treatment with SAFER®, respectively. When body fluids contaminated with ASFV, such as blood, saliva, urine, and feces, were treated with SAFER® for 20  min, the ASFV titer was reduced by 1.6, 2.2, 2.0, and 2.2 Log10HAD50/ml, respectively. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) is a highly contagious and severe hemorrhagic transboundary swine viral disease with up to a 100% mortality rate, which leads to a tremendous socio-economic loss worldwide. The lack of safe and efficacious ASF vaccines is the greatest challenge in the prevention and control of ASF. In this study, we generated a safe and effective live-attenuated virus (LAV) vaccine candidate VNUA-ASFV-LAVL3 by serially passaging a virulent genotype II strain (VNUA-ASFV-L2) in an immortalized porcine alveolar macrophage cell line (3D4/21, 50 passages). VNUA-ASFV-LAVL3 lost its hemadsorption ability but maintained comparable growth kinetics in 3D4/21 cells to that of the parental strain. Notably, it exhibited significant attenuation of virulence in pigs across different doses (103, 104, and 105 TCID50). All vaccinated pigs remained healthy with no clinical signs of African swine fever virus (ASFV) infection throughout the 28-day observation period of immunization. VNUA-ASFV-LAVL3 was efficiently cleared from the blood at 14–17 days post-infection, even at the highest dose (105 TCID50). Importantly, the attenuation observed in vivo did not compromise the ability of VNUA-ASFV-LAVL3 to induce protective immunity. Vaccination with VNUA-ASFV-LAVL3 elicited robust humoral and cellular immune responses in pigs, achieving 100% protection against a lethal wild-type ASFV (genotype II) challenge at all tested doses (103, 104, and 105 TCID50). Furthermore, a single vaccination (104 TCID50) provided protection for up to 2 months. These findings suggest that VNUA-ASFV-LAVL3 can be utilized as a promising safe and efficacious LAV candidate against the contemporary pandemic genotype II ASFV. [ABSTRACT FROM AUTHOR]

African swine fever virus (ASFV) is the causative agent of African swine fever (ASF), a highly infectious and lethal disease of domesticated swine. Outbreaks of ASF have been mostly restricted to the continent of Africa. The outbreaks that have occurred outside of Africa were controlled by extensive depopulation of the domesticated pig population. However, in 2007, an outbreak occurred in the country of Georgia, where ASFV infected wild pigs and quickly spread across eastern Europe. Since the reintroduction of ASF into Europe, variants of the current pandemic strain, ASFV Georgia 2007/01 (ASFV-G), which is classified as Genotype 2 based on p72 sequencing, have been reported in countries within western Europe, Asia, and the island of Hispaniola. Additionally, isolates collected in 2020 confirmed the presence of variants of ASFV-G in Nigeria. Recently, we reported similar variants of ASFV-G collected from domestic pigs suspected of dying of ASF in Ghana in 2022. Here, we retroactively report, based on full-length sequencing, that similar variants were present in Ghana in 2021. The SNP analysis revealed derivatives of ASFV with distinct genetic markers. Furthermore, we identified three full-length ASFV genomes as Genotype 1, indicating that there were two genotypes circulating in proximity during the 2021 ASF outbreaks in Ghana. [ABSTRACT FROM AUTHOR]

Pseudorabies virus (PRV) is one of the herpes viruses that can infect a wide range of animals including pigs, cattle, sheep, mice, and wild animals. PRV is a neurotropic alphaherpesvirus capable of infecting a variety of mammals. There is a rising interest in the targeted application of probiotic bacteria to prevent viral diseases, including PRV. In this study, the surface expression of enhanced green fluorescent protein (EGFP) on recombinant Lactiplantibacillus plantarum NC8 (rNC8) through the LP3065 LPxTG motif of Lactobacillus plantarum WCFS1 was generated. The surface expression was observed through confocal microscopy. Dendritic cell targeting peptides (DCpep) were also fused with LPxTG that help to bind with mouse DCs. The PRV-gD was cloned in LP3065 LPxTG, resulting in the generation of rNC8-LP3065-gD. Inactivated rNC8-LP3065-gD was administered intravenously in mice on days 1 and 7 at a dose of 200 µL (109 CFU/mouse) for monitoring immunogenicity. Subsequently, a challenge dose of PRV TJ (104 TCID50) was administered intramuscularly at 14 days post-immunization. The survival rate of the immunized mice reached 80% (4/5) with no significant signs of illness. A significant rise in anti-gD antibodies was detected in the immunized mice by ELISA. Quantitative PCR (qPCR) results showed decreased viral loading in different body tissues. Flow cytometry of lymphocytes derived from mice spleen indicated an increase in CD3+CD4+ T cells, but CD3+CD8+ T cells were not detected. Moreover, it offers a model to delineate immune correlates with rNC8-induced immunity against swine viral diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine pleuropneumonia, caused by Actinobacillus pleuropneumoniae, is one of the main causes of massive losses in the pig industry, with a high economic impact. This study aimed to identify A. pleuropneumoniae serotypes isolated from pneumonic lesions in dead pigs, naturally infected, in some Piedmontese farms and to describe the macroscopic and microscopic lesions. The identification of circulating serotypes in swine farming is crucial to adopting optimal strategies to control the diseases, such as vaccination plans and biosecurity measures. Porcine pleuropneumonia (PPP) is one of the main causes leading to massive losses in the pig industry, with high economic impacts. Among different etiological agents, Actinobacillus pleuropneumoniae (APP) is responsible for severe fibrinous-necrotizing pleuropneumonia. A total of 19 different APP serotypes are currently recognized. This study aimed to identify APP serotypes isolated from pneumonic lesions in naturally infected and dead pigs in the Piedmont Region and to describe lesions. A total of 107 dead pigs with a suspected PPP diagnosis were included in this study. Lungs were evaluated using gross-pathology scoring systems, histopathology, and APP isolation and serotypes identification by multiplex PCR were conducted. Gross lung lesions were mainly represented by fibrinous pneumonia and pleuropneumonia. APP was isolated in 20/107 (18.7%) samples. PCR indicated APP DNA presence in 53/107 (49.5%) of lung samples. The most observed serotypes were serotype 2 in 24/53 (45.3%) and serotype 6 in 13/53 (24.5%) samples. Moreover, multiplex PCR results suggested a coinfection of different serotypes in five samples. This study emphasizes the importance of an integrated approach, utilizing various techniques, such as gross- and histopathology, and bacteriological culture and PCR, to enhance the diagnosis of APP infections. [ABSTRACT FROM AUTHOR]

The swine acute diarrhea syndrome coronavirus (SADS-CoV) has caused significant disruptions in porcine breeding and raised concerns about potential human infection. The nucleocapsid (N) protein of SADS-CoV plays a vital role in viral assembly and replication, but its structure and functions remain poorly understood. This study utilized biochemistry, X-ray crystallography, and immunization techniques to investigate the N protein’s structure and function in SADS-CoV. Our findings revealed distinct domains within the N protein, including an RNA-binding domain, two disordered domains, and a dimerization domain. Through biochemical assays, we confirmed that the N-terminal domain functions as an RNA-binding domain, and the C-terminal domain is involved in dimerization, with the crystal structure analysis providing visual evidence of dimer formation. Immunization experiments demonstrated that the disordered domain 2 elicited a significant antibody response. These identified domains and their interactions are crucial for viral assembly. This comprehensive understanding of the N protein in SADS-CoV enhances our knowledge of its assembly and replication mechanisms, enabling the development of targeted interventions and therapeutic strategies. IMPORTANCE SADS-CoV is a porcine coronavirus that originated from a bat HKU2-related coronavirus. It causes devastating swine diseases and poses a high risk of spillover to humans. The coronavirus N protein, as the most abundant viral protein in infected cells, likely plays a key role in viral assembly and replication. However, the structure and function of this protein remain unclear. Therefore, this study employed a combination of biochemistry and X-ray crystallography to uncover distinct structural domains in the N protein, including RNA-binding domains, two disordered domains, and dimerization domains. Additionally, we made the novel discovery that the disordered domain elicited a significant antibody response. These findings provide new insights into the structure and functions of the SADS-CoV N protein, which have important implications for future studies on SADS-CoV diagnosis, as well as the development of vaccines and anti-viral drugs. [ABSTRACT FROM AUTHOR]

Porcine circovirus type 3 (PCV3) is closely associated with various diseases, such as the porcine dermatitis, nephropathy syndrome, and multisystemic clinicopathological diseases. PCV3-associated diseases are increasingly recognized as severe diseases in the global swine industry. Ring finger protein 2 (RNF2), an E3 ubiquitin ligase exclusively located in the nucleus, contributes to various biological processes. This ligase interacts with the PCV3 Cap. However, its role in PCV3 replication remains unclear. This study confirmed that the nuclear localization signal domain of the Cap and the RNF2 N-terminal RING domain facilitate the interaction between the Cap and RNF2. Furthermore, RNF2 promoted the binding of K48-linked polyubiquitination chains to lysine at positions 139 and 140 (K139 and K140) of the PCV3 Cap, thereby degrading the Cap. RNF2 knockdown and overexpression increased or decreased PCV3 replication, respectively. Moreover, the RING domain-deleted RNF2 mutant eliminated the RNF2- induced degradation of the PCV3 Cap and RNF2-mediated inhibition of viral replication. This indicates that both processes were associated with its E3 ligase activity. Our findings demonstrate that RNF2 can interact with and degrade the PCV3 Cap via its N-terminal RING domain in a ubiquitination-dependent manner, thereby inhibiting PCV3 replication. IMPORTANCE Porcine circovirus type 3 is a recently described pathogen that is prevalent worldwide, causing substantial economic losses to the swine industry. However, the mechanisms through which host proteins regulate its replication remain unclear. Here, we demonstrate that ring finger protein 2 inhibits porcine circovirus type 3 replication by interacting with and degrading the Cap of this pathogen in a ubiquitination-dependent manner, requiring its N-terminal RING domain. Ring finger protein 2-mediated degradation of the Cap relies on its E3 ligase activity and the simultaneous existence of K139 and K140 within the Cap. These findings reveal the mechanism by which this protein interacts with and degrades the Cap to inhibit porcine circovirus type 3 replication. This consequently provides novel insights into porcine circovirus type 3 pathogenesis and facilitates the development of preventative measures against this pathogen. [ABSTRACT FROM AUTHOR]

The outbreak of infectious diseases in swine, such as classical swine fever (CSF), has become a significant concern in the pig‐farming industry. In Japan, after the re‐emergence of CSF in 2018, farms are now exposed to the risk of transmission from infected wild boar and CSF‐contaminated farms. This study aimed to identify biosecurity measures that were effective for the prevention of CSF introduction into farms during the period from the beginning of the CSF epidemic to the implementation of a vaccination campaign for domestic pigs at risk. The probability of virus introduction was assumed to be increased by the elevated risk from CSF‐infected wild boar and infected farms around the farm. The risk from infected wild boar was represented by the prevalence of CSF in wild boar or the occupancy of 1‐km grid cells with infected wild boar within 10‐km radii from a pig farm and the occurrence of CSF outbreaks on neighboring farms. Conversely, the probability of virus introduction was assumed to decrease in response to on‐farm biosecurity measures being implemented on each farm. The implementation of biosecurity measures on the farms and farm attributes were obtained through a questionnaire survey. Analyses were performed on each farm under the weekly situations where infected wild boar were both absent and present in the vicinity using a binomial generalized linear model. On farms where infected wild boar were not present around farms, daily washing and disinfecting of work clothing in pig houses was identified as the main measure to reduce the risk of CSF introduction into farms. On farms with infected wild boar in the vicinity, the absence of public roads on the farm and preventing wildlife intrusion into the areas where pig carcasses were stored were demonstrated to be effective in preventing CSF introduction. Based on the assumption that strict and comprehensive biosecurity measures are required to prevent CSF introduction, the implementation of these potentially effective measures is worth being prioritized. [ABSTRACT FROM AUTHOR]

• This study presents a novel combined vaccines candidate against FMD and SVA, and provides new insights for the development of combination vaccines for other viral swine diseases. Here we report, to our knowledge, the first development of a VLPs-based combined vaccines targeting FMDV type A and O, and SVA strains. • Inactivated vaccines are unable to serologically differentiate infected from vaccinated animals, thus hindering pathogen eradication. Beneficially, virus-like particles (VLPs)-based vaccines resemble natural viruses in size and antigen structure, offering an alternative tool to circumvent these limitations. • We formulated a combined vaccines consisting of three distinct VLPs specifically designed to target SVA as well as FMDV serotypes O and A. The immunocompatibility and immunological efficacy of this multi-target vaccines were rigorously assessed by optimizing various formulation parameters, immune indicators, and safety metrics. • Pig immunization studies revealed that the combined VLPs vaccines elicited antigen-specific and high serum antibody responses comparable to those elicited by its corresponding individual vaccines. Additionally, the combined VLPs vaccines immune serums strongly neutralized FMDV type A and O, and SVA strains with neutralization titers similar to those of their individual counterparts, suggesting a good compatibility among the three VLPs components regarding their immunogenicity. • Importantly, sera from pigs immunized with the combined VLPs vaccines conferred efficient protection against single or mixed infections with FMDV type A and O, and SVA viruses. In contrast, the individual vaccines could only protect pigs against homologous virus infections and were not effective against heterologous challenges. Inactivated vaccines lack the capability to serologically differentiate between infected and vaccinated animals, thereby impeding the effective eradication of pathogen. Conversely, vaccines based on virus-like particles (VLPs) emulate natural viruses in both size and antigenic structure, presenting a promising alternative to overcome these limitations. As the complexity of swine infectious diseases increases, the increase of vaccine types and doses may intensify the stress response. This exacerbation can lead to diminished productivity, failure of immunization, and elevated costs. Given the critical dynamics of co-infection and the clinically indistinguishable symptoms associated with foot-and-mouth disease virus (FMDV) and senecavirus A (SVA), there is a dire need for an efficacious intervention. To address these challenges, we developed a combined vaccine composed of three distinct VLPs, specifically designed to target SVA and FMDV serotypes O and A. Our research demonstrates that this trivalent VLP vaccine induces antigen-specific and robust serum antibody responses, comparable to those produced by the respective monovalent vaccines. Moreover, the immune sera from the combined VLP vaccine strongly neutralized FMDV type A and O, and SVA, with neutralization titers comparable to those of the individual vaccines, indicating a high level of immunogenic compatibility among the three VLP components. Importantly, the combined VLPs vaccines-immunized sera conferred efficient protection against single or mixed infections with FMDV type A and O, and SVA viruses in pigs. In contrast, individual vaccines could only protect pigs against homologous virus infections and not against heterologous challenges. This study presents a novel combined vaccines candidate against FMD and SVA, and provides new insights for the development of combination vaccines for other viral swine diseases. [ABSTRACT FROM AUTHOR]

African Swine Fever (ASF) is a reportable disease of swine that causes farreaching losses to affected countries and regions. Early detection is critically important to contain and mitigate the impact of ASF outbreaks, for which timely available data is essential. This research examines the potential use of Google Trends data as an early indicator of ASF outbreaks in Southeast Asia, focusing on the three largest swine producing countries, namely, Vietnam, the Philippines, and Thailand. Cross-correlation and Kullback-Leibler (KL) divergence indicators were used to evaluate the association between Google search trends and the number of ASF outbreaks reported. Our analysis indicate strong and moderate correlations between Google search trends and number of ASF outbreaks reported in Vietnam and the Philippines, respectively. In contrast, Thailand, the country of this group in which outbreaks were reported last, exhibits the weakest correlation (KL = 2.64), highlighting variations in public awareness and disease dynamics. These findings suggest that Google search trends are valuable for early detection of ASF. As the disease becomes endemic, integrating trends with other epidemiological data may support the design and implementation of surveillance strategies for transboundary animal diseases in Southeast Asia. [ABSTRACT FROM AUTHOR]

This study aims to explore the current application status and development prospects of Chinese herbal medicine in preventing and treating swine viral infectious diseases over the past five years. By adopting the method of literature review, we collect, organize, and analyze relevant research literature, with the goal of summarizing and summarizing the research progress in this field. Through research, we found that swine infectious diseases have caused serious economic losses in the breeding industry and that some diseases cannot be fully protected by using vaccines. Therefore, we need new means to prevent diseases. The use of Chinese herbal medicine for the prevention and treatment of related diseases has become a reliable means. This study first briefly introduces the common infectious diseases of pigs and the risks and challenges faced by prevention and treatment, then reads the literature comparing the treatment with Western medicine and traditional Chinese medicine, proving the reliability of traditional Chinese medicine treatment and the unique advantages of Chinese herbal medicine. Afterwards, we summarized the literature on the prevention and treatment of related swine diseases with Chinese herbal medicine in the past five years, and finally made a summary and outlook, hoping to provide ideas for relevant researchers and workers. [ABSTRACT FROM AUTHOR]

One of the most significant diseases in the swine business, porcine reproductive and respiratory syndrome virus (PRRSV) causes respiratory problems in piglets and reproductive failure in sows. The PRRSV nucleocapsid (N) protein is essential for the virus' assembly, replication, and immune evasion. Stages in the viral replication cycle can be impacted by interactions between the PRRSV nucleocapsid protein and the host protein components. Therefore, it is of great significance to explore the interaction between the PRRSV nucleocapsid protein and the host. Nevertheless, no information has been published on the network of interactions between the nucleocapsid protein and the host proteins in primary porcine alveolar macrophages (PAMs). In this study, 349 host proteins interacting with nucleocapsid protein were screened in the PRRSV-infected PAMs through a liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based proteomics approach. Bioinformatics analysis, which included gene ontology annotation, Kyoto Encyclopedia of Genes and Genomes database enrichment, and a protein–protein interaction (PPI) network, revealed that the host proteins interacting with PRRSV-N may be involved in protein binding, DNA transcription, metabolism, and innate immune responses. This study confirmed the interaction between the nucleocapsid protein and the natural immune-related proteins. Ultimately, our findings suggest that the nucleocapsid protein plays a pivotal role in facilitating immune evasion during a PRRSV infection. This study contributes to enhancing our understanding of the role played by the nucleocapsid protein in viral pathogenesis and virus–host interaction, thereby offering novel insights for the prevention and control of PRRS as well as the development of vaccines. [ABSTRACT FROM AUTHOR]

The African swine fever virus (ASFV) is an often deadly disease in swine and poses a threat to swine livestock and swine producers. With its complex genome containing more than 150 coding regions, developing effective vaccines for this virus remains a challenge due to a lack of basic knowledge about viral protein function and protein–protein interactions between viral proteins and between viral and host proteins. In this work, we identified ASFV-ASFV protein–protein interactions (PPIs) using artificial intelligence-powered protein structure prediction tools. We benchmarked our PPI identification workflow on the Vaccinia virus, a widely studied nucleocytoplasmic large DNA virus, and found that it could identify gold-standard PPIs that have been validated in vitro in a genome-wide computational screening. We applied this workflow to more than 18,000 pairwise combinations of ASFV proteins and were able to identify seventeen novel PPIs, many of which have corroborating experimental or bioinformatic evidence for their protein–protein interactions, further validating their relevance. Two protein–protein interactions, I267L and I8L, I267L__I8L, and B175L and DP79L, B175L__DP79L, are novel PPIs involving viral proteins known to modulate host immune response. [ABSTRACT FROM AUTHOR]

Shiga toxin-producing Escherichia coli (STEC) poses a significant public health risk due to its zoonotic potential and association with severe human diseases, such as hemorrhagic colitis and hemolytic uremic syndrome. Ruminants are recognized as primary reservoirs for STEC, but swine also contribute to the epidemiology of this pathogen, highlighting the need for effective prevention strategies across species. Notably, a subgroup of STEC that produces Shiga toxin type 2e (Stx2e) causes edema disease (ED) in newborn piglets, economically affecting pig production. This study evaluates the immunogenicity of a chimeric protein-based vaccine candidate against STEC in pregnant sows and the subsequent transfer of immunity to their offspring. This vaccine candidate, which includes chimeric proteins displaying selected epitopes from the proteins Cah, OmpT, and Hes, was previously proven to be immunogenic in pregnant cows. Our analysis revealed a broad diversity of STEC serotypes within swine populations, with the cah and ompT genes being prevalent, validating them as suitable antigens for vaccine development. Although the hes gene was detected less frequently, the presence of at least one of these three genes in a significant proportion of STEC suggests the potential of this vaccine to target a wide range of strains. The vaccination of pregnant sows led to an increase in specific IgG and IgA antibodies against the chimeric proteins, indicating successful immunization. Additionally, our results demonstrated the effective passive transfer of maternal antibodies to piglets, providing them with immediate, albeit temporary, humoral immunity against STEC. These humoral responses demonstrate the immunogenicity of the vaccine candidate and are preliminary indicators of its potential efficacy. However, further research is needed to conclusively evaluate its impact on STEC colonization and shedding. This study highlights the potential of maternal vaccination to protect piglets from ED and contributes to the development of vaccination strategies to reduce the prevalence of STEC in various animal reservoirs. [ABSTRACT FROM AUTHOR]

Swine are increasingly studied as animal models of human disease. The anatomy, size, longevity, physiology, immune system, and metabolism of swine are more like humans than traditional rodent models. In addition, the size of swine is preferred for surgical placement and testing of medical devices destined for humans. These features make swine useful for biomedical, pharmacological, and toxicological research. With recent advances in gene-editing technologies, genetic modifications can readily and efficiently be made in swine to study genetic disorders. In addition, gene-edited swine tissues are necessary for studies testing and validating xenotransplantation into humans to meet the critical shortfall of viable organs versus need. Underlying all of these biomedical applications, the knowledge of husbandry, background diseases and lesions, and biosecurity needs are important for productive, efficient, and reproducible research when using swine as a human disease model for basic research, preclinical testing, and translational studies. [ABSTRACT FROM AUTHOR]

Simple Summary: Diseases in the swine industry can cause major economic and health issues. This review examines R0 estimates, which measure how efficiently respiratory diseases spread among pigs and compares transmission risks within and between farms. Seven diseases were studied: Aujeszky's disease, PPRSV, Circovirus, Influenza A, Encephalomyocarditis Virus, Classical Swine Fever, and African Swine Fever. Data from research articles showed varying R0 values, with higher transmission rates within herds for some diseases. Effective disease control requires prompt detection, response to outbreaks, and continuous monitoring to reduce its impact on the swine industry. Diseases in the swine industry can cause significant economic and health impacts. This review examines R0 estimates for respiratory diseases in pigs, assessing variations and comparing transmission risks within and between farms. A literature search of three databases aggregated peer-reviewed research articles on swine viral respiratory diseases' R0 values. The study focused on seven diseases: Aujeszky's disease (AD), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Circovirus, Influenza A (IA), Encephalomyocarditis Virus (EV), Classical Swine Fever (CSF), and African Swine Fever (ASF). R0 values were estimated for transmission within and between herds/farms using various methods, from complex mathematical models to simple calculations. Data primarily came from disease surveillance and laboratory experiments. The median R0 for within-herd and between-herd transmission was 10 and 3.31 for AD, 2.78 and 1.14 for PRRSV, 5.9 and 0.89 for Circovirus, 1.75 and 1.6 for CSF, and 3.94 and 3.15 for ASF. For IA and EV, only within-herd R0 values were estimated at 8.65 and 1.3, respectively. Diseases with high R0 values highlight the need for prompt detection and response to outbreaks. Continuous monitoring and evaluation of pathogen transmissibility are crucial for enhancing disease surveillance and reducing the impact of livestock diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: ASF remains a significant concern for the Philippines as it has impacted the swine industry and food security. It causes high mortality rates, leading to decreased pork production, high prices of pork and pork products, and financial losses for farmers and stakeholders, especially for the livelihoods of many small-scale pig farmers in the Philippines. Since its first appearance in 2019, the government has implemented strict measures to control the spread of the disease that include enhanced surveillance, strict biosecurity and quarantine protocols, restricted movement of live pigs and pork products, and culling of infected and at-risk pigs. Ongoing surveillance and cooperation between industry stakeholders, government agencies, and research institutions and collaboration with international organizations and experts to develop enhanced strategies for ASF control and prevention are necessary to mitigate ASF's impacts on the swine industry. Moreover, public awareness campaigns and support for affected farmers are continuously in place to combat ASF in the Philippines. African swine fever (ASF), a highly contagious disease of swine, has posed a significant global threat to the swine industry. As an archipelago, the Philippines has a geographic advantage when it comes to the risk of ASF transmission. However, since its introduction to the Philippines in 2019, it has proliferated not only in backyard and commercial farms but also in wild pig populations. While certain parts of the country were more affected than others, the epidemiologic features of ASF necessitate that all affected areas must be closely monitored and that confirmed cases be treated with the utmost care. With the very limited data on ASF epidemiology and surveillance in the Philippines, future efforts to combat ASF must place even greater emphasis on improved prevention and control strategies. It is worth mentioning that the government's efforts toward comprehensive ASF surveillance and epidemiological investigation into the possible ASFV sources or transmission pathways are the most important measures in the prevention and control of ASF outbreaks. This review article provides a comprehensive overview of the current swine industry and ASF situation in the Philippines, which includes its epidemiology, surveillance, prevention, and control strategies. [ABSTRACT FROM AUTHOR]

Wild animals appear to be a key factor in the occurrence, transmission and prevalence of a myriad of contagious animal diseases, being natural reservoirs, vectors or both. This role is played by wild boars in the African swine fever (ASF) transmission to domestic pigs. ASF entered the European Union in 2014 and since then lots of measures have been implemented to bring the disease under control. Bulgaria also tried to introduce new measures for reduction of wild boar populations, regarding ASF through legislative amendments. Proposals in the Bulgarian Law on hunting and game protection from 2020 tried to make legitimate wild boar culling with some unselective approaches like the use of baits with chemical poisonous or intoxicating substances. This provoked the authors to study the experience of other countries on particular toxicants used for baiting for control of wild boar populations. We investigated the available scientific literature on the selective character of chemical baiting and the effect on non-target species. Moreover, the participation of different stakeholders in the process of development and implementation of the mentioned measures was discussed. Thus, proposals were made for better preparatory, scientific and efficiency investigation at the preliminary stage of animal disease control measures development. [ABSTRACT FROM AUTHOR]

Stomach ulcers are relatively common in pigs raised in modern large-scale farms. They can result in reduced daily weight gain, reduced feed intake, and sudden death, resulting in significant economic losses. They also belong to the most common findings in the stomach of pigs at slaughterhouses and are among the most important causes of peracute mortality, associated with bleeding into the digestive system of pigs. The aim of this article was to provide an overview of the most important etiological factors, and pathogenesis of gastric ulcers in pigs. [ABSTRACT FROM AUTHOR]

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Enterotoxigenic Escherichia coli (ETEC) causes post-weaning diarrhea in piglets, significantly impacting animal welfare and production efficiency. The two primary ETEC pathotypes associated with post-weaning diarrhea are ETEC F4 and ETEC F18. During the post-weaning period, piglets may be exposed to both ETEC F4 and ETEC F18. However, the effects of coinfection by both strains have not been studied. Short chain fatty acid feed additives, such as butyrate and valerate, are being investigated for their potential to improve animal performance and disease resistance. Therefore, this pilot experiment aimed to test the effects of butyrate glycerides or valerate glycerides on growth performance, diarrhea incidence, and immune responses of piglets under ETEC F4-ETEC F18 coinfection conditions. Twenty piglets were individually housed and assigned to one of the three dietary treatments immediately at weaning (21 to 24 d of age). The dietary treatments included control (basal diet formulation), control supplemented with 0.1% butyrate glycerides or 0.1% valerate glycerides. After a 7-d adaptation, all pigs were inoculated with ETEC F4 and ETEC F18 (0.5 × 109 CFU/1.5 mL dose for each strain) on three consecutive days. Pigs and feeders were weighed throughout the trial to measure growth performance. Fecal cultures were monitored for hemolytic coliforms, and blood samples were collected for whole blood and serum analysis. Pigs fed valerate glycerides tended (P = 0.095) to have higher final body weight compared with control. The overall severity of diarrhea was significantly (P

Botanicals exhibit promising impacts on intestinal health, immune-regulation, and growth promotion in weaned pigs. However, these benefits may vary depending on major active components in the final feed additive products. Therefore, this study aimed to investigate two types of botanical blends (BB) that were comprised of 0.3% capsicum oleoresin and 12% garlic extracts from different sources on performance, diarrhea, and health of weaned piglets experimentally infected with a pathogenic Escherichia coli F18. Sixty weanling pigs (7.17 ± 0.97 kg body weight (BW)) blocked by weight and gender were assigned to one of five dietary treatments: negative control (NC), positive control (PC), or dietary supplementation with 100 mg/kg of BB1, 50 mg/kg or 100 mg/kg of BB2. This study lasted 28 d with 7 d before and 21 d after the first E. coli inoculation (day 0). All pigs, except negative control, were orally inoculated with 1010 cfu E. coli F18/3-mL dose for 3 consecutive days. Blood samples were collected periodically to analyze systemic immunity. Intestinal tissues and mucosa were collected on days 5 and 21 PI for analyzing histology and gene expression. All data, except for frequency of diarrhea, were analyzed by ANOVA using the PROC MIXED of SAS. The Chi-square test was used for analyzing frequency of diarrhea. Escherichia coli infection reduced (P < 0.05) growth rate and feed intake and increased (P < 0.05) frequency of diarrhea of weaned pigs throughout the experiment. Supplementation of 100 mg/kg BB1 or BB2 alleviated (P < 0.05) frequency of diarrhea of E. coli challenged pigs during the entire experiment. Escherichia coli infection also enhanced (P < 0.05) serum TNF-α and haptoglobin concentrations on day 4 post-inoculation (PI) but reduced (P < 0.05) duodenal villi height and area on day 5 PI, while pigs supplemented with 100 mg/kg BB1 or BB2 had lower (P < 0.05) serum TNF-α than pigs in PC on day 4 PI. Pigs fed with 100 mg/kg BB2 had higher (P < 0.05) jejunal villi height than pigs in PC on day 5 PI. Pigs fed with 100 mg/kg BB2 had reduced (P < 0.05) gene expression of IL1B, PTGS2, and TNFA in ileal mucosa than pigs in PC on day 21 PI. In conclusion, dietary supplementation of botanical blends at 100 mg/kg could enhance disease resistance of weaned pigs infected with E. coli F18 by enhancing intestinal morphology and regulating local and systemic immunity of pigs.

Background: Monitoring of infectious diseases on swine farms requires a high diagnostic sensitivity and specificity of the test system. Moreover, particularly in cases of swine influenza A virus (swIAV) it is desirable to include characterization of the virus as precisely as possible. This is indispensable for strategies concerning prophylaxis of swIAV and furthermore, to meet the requirements of a purposeful monitoring of newly emerging swIAV strains in terms of vaccine design and public health. Within the present cross-sectional study, we compared the diagnostic value of group samples (wipes of surfaces with direct contact to mouth/nose, dust wipes, udder skin wipes, oral fluids) to individual samples (nasal swabs, tracheobronchial swabs) for both swIAV identification and characterization. Sampling included different stages of pig production on 25 sow farms with attached nursery considered as enzootically infected with swIAV. Firstly, samples were analyzed for IAV genome and subsequently samples with Ct-values < 32 were subtyped by multiplex RT-qPCR. Results: Nasal swabs of suckling piglets and nursery pigs resulted in a higher odds to detect swIAV (p < 0.001) and to identify swIAV subtypes by RT-qPCR (p < 0.05) compared to nasal swabs of sows. In suckling piglets, significant higher rates of swIAV detection could be observed for nasal swabs (p = 0.007) and sow udder skin wipes (p = 0.036) compared to contact wipes. In the nursery, group sampling specimens were significantly more often swIAV positive compared to individual samples (p < 0.01), with exception of the comparison between contact wipes and nasal swabs (p = 0.181). However, in general nasal swabs were more likely to have Ct-value < 32 and thus, to be suitable for subtyping by RT-qPCR compared to dust wipes, contact wipes, udder skin wipes and tracheobronchial swabs (p < 0.05). Interestingly, different subtypes were found in different age groups as well as in different specimens in the same holding. Conclusion: Although population-based specimens are highly effective for swIAV monitoring, nasal swabs are still the preferable sampling material for the surveillance of on-farm circulating strains due to significantly higher virus loads. Remarkably, sampling strategies should incorporate suckling piglets and different age groups within the nursery to cover as many as possible of the on-farm circulating strains. [ABSTRACT FROM AUTHOR]

Human health is dependent on food safety and, therefore, on the health of farm animals. One of the most significant threats in regard to swine diseases is African swine fever (ASF). Infections caused by porcine circoviruses (PCVs) represent another important swine disease. Due to the ubiquitous nature of PCV2, it is not surprising that this virus has been detected in ASFV-affected pigs. However, recent data indicate that coinfection of PCV3 and ASFV also occurs. It is still unclear whether PCV infection plays a role in ASFV infection, and that subject requires further analysis. The aim of this study was to assess whether PCV3 and PCV4 are present in the wild boar population in Poland (real-time PCR). The analysis was performed on wild boar samples collected for routine ASF surveillance in Poland, between 2018 and 2021. By extension, the obtained data were compared in regard to ASFV presence in these samples, thus investigating the odds of ASFV infection on the grounds of the PCV carrier state in free-ranging Suidae in Poland. In addition, sequencing of PCV3 and phylogenetic analysis were performed, based on a full genome and a capsid gene. In the current study, we demonstrated the high prevalence of PCV3 in the wild boar population in Poland; meanwhile, PCV4 was not detected. The odds of ASFV infection on the grounds of the PCV3 carrier state in free-ranging Suidae in Poland was more than twice as high. Ten full genome sequences of PCV3 were obtained, all of them belonging to clade 3a. The similarity between them was in the range of 98.78–99.80%. [ABSTRACT FROM AUTHOR]

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African swine fever (ASF) is a deadly disease of swine currently causing a worldwide pandemic, leading to severe economic consequences for the porcine industry. The control of disease spread is hampered by the limitation of available effective vaccines. Live attenuated vaccines (LAVs) are currently the most advanced vaccine prototypes, providing strong protection against ASF. However, the significant advances achieved using LAVs must be complemented with further studies to analyze vaccine-induced immunity. Here, we characterized the onset of cross-protective immunity triggered by the LAV candidate BA71ΔCD2. Intranasally vaccinated pigs were challenged with the virulent Georgia 2007/1 strain at days 3, 7 and 12 postvaccination. Only the animals vaccinated 12 days before the challenge had effectively controlled infection progression, showing low virus loads, minor clinical signs and a lack of the unbalanced inflammatory response characteristic of severe disease. Contrarily, the animals vaccinated 3 or 7 days before the challenge just showed a minor delay in disease progression. An analysis of the humoral response and whole blood transcriptome signatures demonstrated that the control of infection was associated with the presence of virus-specific IgG and a cytotoxic response before the challenge. These results contribute to our understanding of protective immunity induced by LAV-based vaccines, encouraging their use in emergency responses in ASF-affected areas. [ABSTRACT FROM AUTHOR]