Brief Summary of Minutes of Annual Meeting

The Business meeting started with an introduction from the NC229 Chair, Dr Pineiro, summarizing the overaching goal and the objectives of the project.

The meeting proceeded with five 15-minute presentation from five station representatives (or personnel from their team): Illinois, North Carolina, Minnesota, Nebraska, and Kentucky.

Lastly, the NC229 Secretary, Dr Arruda, gave an overview of the information captured through this report, and updated the group on on-going discussions regarding future meeting time and location. A survey was made available for all meeting attendees to contribute their preferences. Lastly, the station reps voted to elect a new Member-at-Large member, namely Dr Mariana Kikuti from the University of Minnesota.

Short-Term Multi-State Outcomes

Objective 1: Etiology, pathogenesis, and virus-host interaction

1a. Virus evolution, mechanisms of tropism and virulence

• Global SVA Genetic Diversity & Control Strategies (NY + Federal Univ. of Pelotas, Brazil): New York collaborated with Brazil to characterize Senecavirus A (SVA) genetic diversity, informing improved global control strategies and surveillance priorities.
• Determinants of SVA Virulence & Persistence (NY): NY identified SVA virulence determinants and mechanisms of persistence, supporting development of safer vaccine candidates for industry use.
• PRRSV Genome Variation in U.S. Outbreaks (UMN): Minnesota investigated genome variation in highly pathogenic PRRSV from U.S. outbreaks, enhancing insight into variant emergence and spread.
• Predictive Modeling of PRRSV Spread (UMN): UMN developed an algorithm to predict which PRRSV genetic variants are likely to become more widespread, enabling proactive risk management.
• Experimental Evidence of Vaccine-Driven Viral Evolution (UMN): UMN published data demonstrating that PRRSV vaccine use can accelerate viral evolution across pig-to-pig infection chains, underscoring the need for adaptive strategies.
• PEDV Evolutionary Trends & Recombination (UMN): UMN quantified current evolutionary trends of PEDV in the U.S., identifying whole-genome recombination and two dominant contemporary clades.
• PoAStV4 proof of primary pathogen (NCSU, UC Santa Cruz, USDA-ARS): reproduction of respiratory lesions in CDCD pigs inoculated with PoAstV4 positive tissue homogenate.

1b. Viral gene-protein structure and function, replication

• ASFV p30 Immunogenic Epitope Mapping (NY): NY demonstrated that ASFV p30 is highly immunogenic and identified a dominant epitope responsible for most antibody responses—advancing prospects for a DIVA marker vaccine with major industry benefits.
• PRRSV Glycoprotein Structural Diversity & Neutralization (UMN): UMN showed that structural diversity in glycoproteins did not significantly influence cross-neutralization between wild-type PRRS strains.
• Bacteria-Free PRRSV Reverse Genetics System (UIUC): Illinois developed a simple, fast reverse genetics system for PRRSV that avoids bacterial amplification, improving genetic stability and construction fidelity for PRRSV mutants in vitro.
• Interferon Antagonism & Host Tropism in PDCoV (UIUC): UIUC identified multiple PDCoV proteins that counteract host interferon responses, providing mechanistic insight into broad host range and informing antiviral design.

1c. Host mechanisms associated with resistance, resilience, and susceptibility

• Host Signatures of PRRSV Infection & Persistence (IL + ISU): Illinois, in collaboration with Iowa State, identified host signatures associated with PRRSV infection and persistence to inform biomarkers and intervention targets
• Mechanisms of PRRSV Virulence Variation (IL + ISU + Midwest Practitioners): IL is collaborating with ISU and swine practitioners across Midwest states to elucidate determinants underlying variation in PRRSV virulence in field settings.
• Flow Cytometry Assay for Infection & Necroptosis (SDSU): South Dakota State University established a double-staining flow cytometry protocol to simultaneously detect PRRSV infection and cell death, with published procedures enabling detailed necroptosis analyses in porcine macrophages.
• Pathogenic mechanism of PRRSV-2 NC strains (NCSU): NCSU evaluated the pathogenic mechanism of different NC PRRSV-2 strains by focusing on the alteration of mitochondrial function in primary lung macrophages during early infection and highlighted the response of different macrophage populations to distinct NC PRRSV-2 strains.
• Characterization of swine trophoblast organoids (NCSU + Duke + Kentucky University): NCSU collaborated with Duke and KU for the first characterization of swine trophoblast organoids model and identified novel markers for porcine uterus and placenta using omics approaches. This new organoid model will be used to study PRRSV-placenta interactions and mechanism of vertical transmission.

Objective 2: Epidemiological investigation of viral pathogens affecting the U.S. swine population

2a. Diagnostics and surveillance

• Metagenomics & Targeted WGS Assays (NY + KS; NYS VDL): NY collaborated with Kansas to develop viral metagenomics and targeted whole-genome sequencing assays for SVA, FMDV, ASFV, and CSFV; these were implemented at NYS VDL to enhance preparedness and early detection.
• ASF Diagnosis and Reporting (KS + private industry): Kansas State University and an App developer in Colorado developed a “Rapid, Sensitive, User-friendly, and Field-deployable AI Tool for Enhancing African Swine Fever Diagnosis and Reporting”.
• Emergence of Strain 1H.18 (UMN + Industry/Veterinarians): UMN, with production systems and swine veterinarians, detected, reported, and characterized a newly emergent strain, 1H.18.
• Global ASF Surveillance Network (UMN + Swine Industry): Through the Swine Disease Global Surveillance project, UMN monitored the evolution and global spread of ASF with industry collaboration.
• Smartphone-Linked Field Test for ASFV (IL + KS): Illinois and Kansas validated a smartphone-linked, field-deployable ASFV diagnostic in BSL-3, supporting rapid response capacity.
• DNA Aptamer Nanosensor for PEDV/PDCoV (IL + ISU): Illinois and Iowa State validated a DNA aptamer-based nanosensor for on-farm detection of PEDV and PDCoV.
• Portable POC PCR for Influenza (OSU): OSU optimized avian and mammalian influenza primers (including highly pathogenic strains) for the Kalix portable RT-PCR system and benchmarked sensitivity relative to standard lab RT-PCR—accelerating field diagnostics.
• Processing Fluids/Tongue Tips Pooling Strategies (UMN): UMN evaluated how pooling processing fluids and tongue tips and aggregating litters influence PRRSV detection in breeding herds.
• Viability PCR for PRRSV in Grow-Finish Farms (UMN): UMN assessed detection of viable PRRSV using a recently developed viability PCR test in growing pig farms.
• Near-Real-Time National Variant Reporting (UMN): UMN built a national surveillance report to identify, flag, and track variants with indicators of wider spread, linking to near-real-time site-level data.
• PEDV & Swine IAV in Growing Pigs (UMN): UMN investigated occurrence patterns of PEDV and swine influenza A virus in growing pigs to inform control measures.
• Avian Influenza Investigations in Bangladesh (UMN): UMN supported investigations of H5N1 and H9N2 in live bird markets and among market workers in Bangladesh, reinforcing One Health surveillance.
• Enriched Long-Read Sequencing for Co-Circulating Viruses (UMN): UMN developed and optimized an enriched long-read sequencing approach to resolve co-circulating viruses in pigs.
• PRRSV in Air Emissions from Grow-Finish Farms (UMN): UMN reported detection of PRRSV in farm air emissions, informing airborne transmission mitigation.
• Post-Mortem Blood Sampling Method for PRRS RT-PCR (UMN): UMN validated a new methodology to obtain post-mortem blood for PRRS RT-PCR to expand diagnostic windows.

2b. Spatial-temporal pattern and risk factors

• Midwest Swine Movement & Risk (OH + MI): Ohio and Michigan are collaborating to map movements across commercial, exhibition, and small-scale sectors to improve spread risk estimates and mitigation planning.
• ASF Outbreak Dynamics Abroad (UMN + National Veterinary Services): UMN evaluated ASF outbreaks and drivers in the Philippines and the Dominican Republic in collaboration with official veterinary services.
• SVA Incidence Trends in U.S. Breeding Herds (UMN): UMN described temporal and regional distribution of SVA incidence across the past decade.
• PRRSV Cross-Border Spread (UMN): UMN analyzed PRRS spread dynamics between Canada and the U.S., identifying pathways and timelines.
• PRRSV Variant Emergence & Regional Lag Times (UMN): UMN estimated lag times between variant emergence in one region and dispersal to others to inform preparedness windows.
• NC PRRSV-2 strains evolution (NCSU+UMN): NCSU collaborated with UMN for NC PRRSV-2 strains evolution, classification and by sharing NC PRRSV-2 strains. NCSU also collaborated with Cambridge Technologies (MN) for whole genome sequencing and NC PRRSV-2 annotations in NCBI GenBank.

2c. Outbreak investigations

• PRRS Outbreak Source Analysis (OH + Neighboring states): Ohio conducted PRRS outbreak investigations (including opportunities extended to IN, PA, etc.) to identify farm-specific risks and mitigation strategies.
• Microevolution Driving Clinical Re-Breaks (UMN): UMN evaluated the relationship between clinical PRRSV re-breaks and whole-genome microevolution on previously exposed farms.
• MSHMP Network Sequence Analytics (UMN): UMN continues sequence analysis of PRRSV outbreaks via the MSHMP network, enhancing situational awareness.
• Characterizing Emerging PRRS Variant 1C.5.32 (UMN): UMN acquired funding to characterize the epidemiology and immunology of PRRS variant 1C.5.32 causing widespread Midwest outbreaks. 

2d. Disease prevention, control, and elimination

• Producer Education on ASF/FMD in Puerto Rico (OH + PR): Ohio collaborated with Puerto Rico to educate producers on ASF/FMD transmission risks, improving early recognition and reducing future spread risk on the island.
• Toward Recombination-Resistant PRRS Vaccines (IL + OH): Illinois and Ohio demonstrated potential resistance of PRRSV to genomic RNA recombination—informing design of recombination-resistant vaccine candidates.
• Biosecurity Gaps in Carcass Management & Rendering (UMN): UMN characterized biosecurity vulnerabilities related to dead animal management and rendering for PRRSV and PEDV.
• HPAI H5N1 Threats & One Health Gaps (UMN): UMN reported emerging threats of HPAI H5N1 clade 2.3.4.4b in swine and highlighted knowledge gaps in One Health contexts.
• Electrostatic Precipitation to Reduce Airborne Virus (UMN): UMN evaluated an electrostatic precipitator to remove viruses from air, mitigating transmission risk.
• Aerosol Particle Size & Viral Load During H1N1 Infection (UMN): UMN quantified the size distribution and viral load of influenza virus–laden airborne particles emitted from pigs over the course of H1N1 infection.
• Trailer Contamination at Harvest Plants (UMN): UMN confirmed market hog trailers become contaminated (PRRSV, coronaviruses, Senecavirus) during unloading at harvest plants—informing cleaning/disinfection protocols.
• PRRS Incidence in Filtered vs. Unfiltered Herds (UMN): UMN quantified PRRS incidence over 15+ years in filtered and unfiltered breeding herds to guide investment in air filtration.
• The Rapid AccessBiosecurity (RAB) app™ (NCSU) has significantly expanded its footprint, now benefiting 36 states and utilized by 121 companies. The platform currently hosts data for over 9,946 swine farms, with 7,325 farms having active, standardized Secure Pork Supply (SPS) biosecurity plans. This widespread adoption allows State Animal Health Officials (SAHOs) to visualize biosecurity infrastructure and compliance in real-time, drastically improving the U.S. swine industry's readiness for Foreign Animal Diseases (FADs).
• NCSU+ SHIC completed a cost-benefit analysis of vehicle rerouting strategies. This work provided the industry with actionable data on how rerouting live-haul vehicles can minimize the dissemination of endemic and emerging diseases between farms. 

2e. Production/Economic impact

• Cost of PRRSV to U.S. Swine Industry (UMN + ISU): Using MSHMP data, UMN contributed to quantifying the economic burden of PRRSV nationally, supporting cost–benefit analyses of interventions.
• Vaccine needs (KSU, ISU, industry partners): Kansas State University worked with experts at Iowa State University and Carthage Veterinary Service in Illinois to determine the types of swine viral and bacterial vaccines that need to be developed in order to reduce the use of antimicrobials used on swine farms to control bacterial infections.

Objective 3: Immunology, vaccinology, and antiviral drug development

3a. Immunology

• ASFV Antigen Discovery for Subunit Vaccines (NY + KS): NY identified and characterized several immunogenic ASFV proteins and is leveraging these findings to design subunit vaccine formulations in collaboration with Kansas.
• T-Cell Epitope Landscape Evolution (UMN + EpiVax): In collaboration with EpiVax, UMN quantified how T-cell epitope landscapes evolve across space and time.
• Neutralizing Antibody Correlates & Protection (UMN): UMN continued investigations into mechanisms of immune protection and correlates of immunity, with emphasis on neutralizing antibodies.
• Machine Learning for PRRSV2 Cross-Reactivity (UMN): UMN developed an in silico ML algorithm predicting cross-reactivity from PRRSV2 genetic sequences to guide vaccine and antibody design.
• GP5-Specific Antibody Profiling (UMN): UMN analyzed GP5-targeted antibody responses against homologous and heterologous viruses to inform breadth of immunity.
• PRRSV-2 immunity in the context of the porcine respiratory disease complex (PRDC) (NCSU+ISU): NCSU collaborated with ISU VDL to conduct a comprehensive study correlating both the anti-PRRSV immune response and 21 secondary infectious agents involved in the severity of PRDC. The study underscored the potential of NanoString technology for comprehensive pathogen surveillance.
• PRRSV-Vit D research (NCSU+ISU): NCSU is collaborating with ISU to study the impact of dietary vitamin D on both acute PRRSV infection as well as immune response to PRRSV MLV vaccination.

3b. Vaccinology

• ASFV Vaccine Candidate Evaluation (NY + KS): NY and Kansas are assessing immunogenicity and protective efficacy of ASFV vaccine candidates
• Broadly Protective Influenza Platforms (NY + OH): NY collaborated with Ohio to develop and evaluate broadly protective vaccine platforms for swine influenza—benefiting animal and public health.
• Broad mRNA Vaccines Against Swine IAV (IL + SDSU + KS): Illinois, South Dakota, and Kansas collaborated to develop broadly protective mRNA vaccines against swine influenza A viruses.
• Custom Influenza Vaccines Reduce Weaning Prevalence (UMN): UMN reported reduced IAV prevalence in pigs at weaning following deployment of custom-made influenza vaccines in breeding herds of an integrated system.
• M2-Based Influenza Vaccine Development (IL + UIUC Biochemistry): IL labs are collaborating with Dr. Stephen Sligar (UIUC Biochemistry) to develop a broadly protective swine influenza vaccine based on Matrix protein 2 (M2).
• Highly Efficacious Swine Influenza Vaccine Prototype (UNL): UNL developed a new, safe, and highly efficacious vaccine prototype for swine influenza, advancing control options.
• PRRSV Molecular Epidemiology & Fitness–Virulence Tradeoffs (IL + MN): Illinois and Minnesota are jointly studying molecular epidemiology and fitness–virulence tradeoffs across pathogen, pig, and population scales for PRRSV variants.
• PRRSV Tropism Insights (UNL): Research at the University of Nebraska–Lincoln provided new insights into PRRSV biology, including tropism, informing novel intervention development.
• PRRSV Vaccine Candidate (KSU, Animal Disease Center, USDA-ARS-Beltsville Animal Research Center, and Tennessee State University): The team performed a swine efficacy study where replication-competent recombinant PRRSV expressing Interferon- ω5 was shown to be an effective modified live virus vaccine against NADC-34 challenge. This work provides a new vaccine candidate to improve swine health.
• NCSU established a PRRSV-2 Immune Biobank for vaccine immunogenicity prediction using In Vitro and In Silico methods (NCSU+ EpiVax): The PRRSV-2 immune biobank demonstrated potential as a tool for predicting vaccine immunogenicity against different NC PRRSV-2 strains and EpiCC (EpiVax) provided additional information on T cell epitope cross conservation

3c. Antiviral drugs

• Evaluation of different organic and inorganic selenium compounds as antivirals and immunomodulators (NCSU): NCSU evaluated the differential effects of organic and inorganic selenium compounds on mitochondrial function in PRRSV-infected porcine alveolar macrophages and their antiviral capacity against PRRSV-2. Selenium treatment did not reduce PRRSV-2 viral replication in porcine alveolar macrophages. Organic DL-selenomethionine exerted beneficial effects on mitochondrial function, and the potential for reducing oxidative stress.

Outputs

• Foreign Animal Disease Preparedness for Viral Diseases of Concern: Ohio State University collaborated with the University of Puerto Rico, Ana Mendez School of Veterinary Medicine, USDA, and Puerto Rico Department of Agriculture to deliver workshops and a tabletop exercise on ASF and FMD across the island. Data collected during these activities will inform future reports and publications.
• ASFV and CSFV Whole Genome Sequencing (NY + KS): NY partnered with Kansas to develop targeted whole genome sequencing methods for ASFV and CSFV using the MinION portable platform (published work). This innovation enhances rapid genomic characterization during outbreaks.
• Influenza Immunology Collaboration (NY + OH): NY collaborated with Ohio to characterize effector antibody functions elicited by influenza A immunization in pigs (published work). Findings inform vaccine design and immune response optimization.
• Cross-Species Influenza Susceptibility (NY + KS): NY and Kansas investigated swine susceptibility to bovine-derived HPAI H5N1, publishing results in Emerging Microbes and Infections (2025). This work informs One Health risk assessments for emerging influenza strains.
• Broadly Protective Influenza Vaccines (UIUC): Illinois constructed candidate antigens and delivery vehicles for swine influenza vaccines, published efficacy data, and filed a patent on this technology.
• PRRSV Variant Classification System (UMN + ISU + SDSU + KSU): UMN collaborated with diagnostic labs to adopt and maintain a PRRSV-2 variant classification system, updated quarterly. UMN also produces monthly reports on variants requiring close monitoring and hosts a web platform for sequence classification.
• PRRSV Persistence Mechanisms (UIUC): Illinois identified candidate host cell types, differentially expressed genes, and viral mutations associated with PRRSV persistence, informing strategies to reduce chronic infection.
• PRRSV Virulence Biotyping (UIUC): Illinois is characterizing highly virulent PRRSV strains (Lineage 1, Type 2) and submitted a progress report to SHIC.
• Nebraska Advances in PRRSV Biology and Vaccinology: Nebraska demonstrated PRRSV infection of spermatogonial stem cells and peritoneal macrophages, expanding knowledge of viral tropism. The station also established a cost-effective Oxford Nanopore sequencing protocol for complete PRRSV genomes and developed a lipid nanoparticle-encapsulated DNA vaccine against swine influenza, effective even in piglets with maternal antibodies.
• Senecavirus A Research and Publications (NY): New York submitted multiple manuscripts currently under review, including studies on SVA genetic diversity in Brazil and the impact of stem-loop I mutations on replication and virulence. These outputs will provide crucial information to advance global understanding of SVA evolution and pathogenicity.
• National Monitoring Programs (UMN): The University of Minnesota continues leadership in the Morrison Swine Health Monitoring Project (MSHMP), covering 55% of the U.S. breeding herd and 12% of the growing pig herd. Weekly epidemiological reports reach hundreds of stakeholders nationwide.
• Next-Generation Diagnostic Tools (UIUC): Illinois advanced biosensor-based diagnostics, developing two platforms: a smartphone-linked ASFV field test and a DNA aptamer nanosensor for PEDV and PDCoV detection on farms. These tools enable rapid, on-site disease detection.
• Bacterial vaccine needs (KSU + ISU + University of Missouri + industry partners): A peer reviewed paper on the bacterial vaccines that need to be developed to control bacterial infections in swine farms
• Field-Deployable App (KSU + App developer in Colorado): A patent application was filed for a "Rapid, Sensitive, User-friendly, and Field-deployable Diagnostic App for ASF Control" (KSURF Disc. NO.: 2025-019; Provisional Patent Application No.: 63/704,903).
• Comparative platforms for Influenza (KSU + ISU): Development of Porcine Air-Liquid Interface Organoids as a comparative platform to study Influenza A Virus infection
• Annotation of 8 whole genome sequences of NC PRRSV-2 isolates (NCSU+UMN+ Cambridge technologies). This has enabled more precise spatial-temporal tracking of PRRSV-2 evolution within NC swine production region and US.
• PRRSV immunity in the context of PRDC (NCSU+ISU): Identification of PCMV and PRRSV-2 as key drivers of clinical severity, together with PPIV-1, B. Bronchiseptica and M. hyorhinis. The approach provides producers with specific targets for multifaceted disease management. Molecular tests performed at ISU VDL had significant positive correlation with NanoString data, underscoring the potential of NanoString technology for comprehensive pathogen surveillance.
• Establishment of the swine trophoblast organoids model for studying viral vertical transmission and reproductive pathology in pigs (NCSU+KU+Duke).
• Establishment of the PRRSV-2 Immune Biobank for prediction of PRRSV vaccine immunogenicity (NCSU+EpiVax). By combining in vitro assays and in silico T cell epitope mapping (EpiCC), the project provides a measurable framework for predicting vaccine immunogenicity against diverse PRRSV field strains in US.
• PoAstV4 proof of primary pathogen (NCSU, UC Santa Cruz, USDA-ARS): reproduced respiratory lesions in CDCD pigs inoculated with PoAstV4 positive tissue homogenate. A shedding curve has been defined as well as the humoral immune response to infection.
• Software (RABapp™)(NCSU): A licensed technology (NCSU innovation number 2021-137) now used by 36 states to map and manage biosecurity plans for the swine industry, covering over 9,900 swine premises.
• Software (MHASpread) (NCSU): An open-source R package (version 0.1.0) for multi-host animal disease spread modeling. Available at: https://github.com/machado-lab/MHASpread-model
• Reports/Data (NCSU+ Missouri): Standardized biosecurity plans for swine producers in Missouri (NADPRP funded), creating a harmonized data structure for interstate disease response.

Multi-State Activities

• ASFV Detection in Boars and Semen (SDSU + ISU): South Dakota and Iowa State investigators completed an ASFV challenge study at Canada’s National Centre for Foreign Animal Diseases to identify optimal sample types and PCR protocols for early ASFV detection in boars, semen, oral fluids, and blood. Results are under evaluation for publication.
• ASF Diagnosis Research (KSU + App developer): Kansas State University and App developer in Colorado developed a deep learning diagnostic tool for image analysis of a lateral flow assay is described. The tool employs an artificial intelligence model that is trained for image classification of images of lateral flow assays. Images of completed lateral flow assays are captured by a camera carried by a mobile device and delivered to the model for image classification as either positive or negative. The results can then be displayed on the mobile device, including the generation of a map that displays the locations of positive results.
• ASFV Protective Antigen Research (NY + KS): New York and Kansas are investigating ASFV proteins involved in protective immune responses to inform subunit vaccine design.
• PRRSV Genetic Variability Analysis (SDSU): SDSU analyzed 225 contemporary PRRSV field strains from the Upper Midwest, identifying L1C.5 as the most prevalent variant in Minnesota and South Dakota. Findings revealed multiple nonsynonymous mutations in immunogenic regions, likely contributing to immune evasion and vaccine escape.
• Novel PRRSV Vaccine Development (SDSU + UMN): SDSU and UMN developed a PRRSV vaccine candidate using a Pichinde virus (PICV) vector. Animal trials demonstrated strong neutralizing antibody responses, reduced viremia and shedding, and less severe lung lesions compared to controls. Cytokine profiling indicated controlled inflammation and enhanced Th1-mediated clearance, positioning this platform for multivalent vaccine development.
• PRRSV Variant Classification and Nomenclature (UMN + ISU + USDA): UMN updates the PRRSV classification system quarterly with new sequence data from MSHMP and collaborates with ISU and USDA to establish a PRRSV-2 nomenclature advisory group for global diversity tracking.
• PRRSV Biotyping and Influenza Vaccine Optimization (Illinois): Illinois optimized methods to determine PRRSV biotypes and conducted pig trials to refine influenza A vaccine formulations using M2 protein nanodiscs.
• PRRSV Tropism and Sequencing Advances (Nebraska): Nebraska identified viral determinants of PRRSV tropism, established a cost-effective Oxford Nanopore sequencing protocol for whole-genome assembly, and developed a lipid nanoparticle-encapsulated DNA vaccine against swine influenza, effective even in piglets with maternal antibodies.
• Influenza A virus evolution, mechanisms of tropism and virulence (KSU + ISU): Kansas State University in collaboration with Iowa State University used a novel ALI-PREC model to comparatively investigate the infection dynamics and early host gene expression responses to human-, swine-, and avian-lineage IAVs. The objectives include identifying key determinants of productive infection, characterizing lineage-specific host responses, and exploring host factors that modulate viral replication.
• Systems Vaccinology Research (KSU + USDA-ARS-National Animal Disease Center, USDA-ARS-Beltsville Animal Research Center + Tennessee State University): The large team leveraged systems vaccinology and transcriptomic profiling to evaluate the swine immune responses resulting from vaccination with prototype PRRSV vaccines that co-express Type 1 interferons (IFNs). We hypothesized that these IFNs would enhance antiviral immunity and improve vaccine efficacy.
• Next-Generation Sequencing Leadership (NY): Cornell advanced genomic surveillance by developing and validating viral metagenomics and targeted whole-genome sequencing assays. SOPs created in collaboration with KS, TX, and CA were distributed to NAHLN labs, with training provided to over 20 laboratories nationwide.
• Reagent and Diagnostic Support (SDSU): SDSU continued providing monoclonal antibodies and diagnostic assay support to collaborating universities, stations, and industry partners for research and diagnostic applications.
• National Disease Monitoring (UMN): The Morrison Swine Health Monitoring Project (MSHMP) continues to deliver weekly disease occurrence reports to industry stakeholders, covering 55% of U.S. breeding herds and supporting preparedness for foreign animal disease emergencies.
• Field-Deployable Diagnostics (Illinois + KS + ISU): Illinois developed a smartphone-linked ASFV test validated by Kansas in BSL-3 labs and a DNA aptamer nanosensor for PEDV and PDCoV detection, with Iowa State providing field samples and qRT-PCR validation.
• Advanced Genomic and Vaccine Research (Illinois + SDSU + KS): Illinois performed animal experiments and scRNAseq, with Iowa State conducting bioinformatics and full-genome analysis. Illinois and SDSU designed vaccine constructs, Illinois generated constructs for testing, and Kansas synthesized nanoparticles for vaccine delivery.
• NanoString technology for comprehensive pathogen surveillance in field conditions (NCSU + ISU): validation of NanoString technology in bronchoalveolar lavage and comparison with VDL molecular tests.
• PRRSV – Vitamin D research (NCSU+ISU): NCSU is studying the impact of dietary vitamin D on both acute PRRSV infection as well as the immune response to PRRSV MLV vaccination. ISU supports these efforts by carrying out PRRV neutralizing antibody assays as well as the growth and titration of challenge virus. NC State handles the running of pig experiments as well as cellular and humoral immune analysis and pathology interpretation.
• International Training Workshops (NCSU- South America): Conducted capacity-building
• workshops on the use of MHASpread for Animal Health Officials from Chile, Panama, El Salvador, Brazil, and Bolivia. These workshops fostered international collaboration on transboundary disease modeling relevant to ASF and FMD.
• NCSU collaboration with State Officials: Ongoing direct collaboration with State Animal Health Officials in 36 states (including key swine producing states like IL, TX, OK, MN, NE, SD, PA,and NC) to implement and refine the RABapp system for real-time biosecurity mapping and movement permitting of swine premises.

Milestones

• Swine Movement Analysis Completed (Ohio + Michigan): Ohio researchers, in collaboration with Michigan, have completed data collection on swine movements—including commercial operations and small producers. Analysis is underway for dissemination of results.
• Vaccine Testing in Animal Models (Illinois + SDSU): Illinois will collaborate with South Dakota to complete testing of vaccine constructs in animal models.
• ASF Workshop and Swine Disease Conferences (KSU, ISU, IL, OSU): KSU in collaboration with Iowa State University and University of Illinois organized one international ASF workshop. KSU in collaboration with Illinois and Ohio State University obtained funding for different conferences on disease infection in pigs and other species.
• ASFV Protective Protein Identification (NY + KS): New York and Kansas completed an immunization challenge study to identify ASFV proteins critical for protective immunity, informing subunit vaccine development.
• ASFV Vaccine Development (KSU + international collaborators): KSU in collaboration with international collaborators developed and tested two safe and efficacious vaccines for ASF.
• ASFV Biosensor Assay Validation (Illinois + KS): Illinois researchers will collaborate with Dr. Jishu Shi (Kansas) to complete validation of biosensor assays for ASFV detection in BSL-3 laboratories.
• ASF Detection (KSU, USDA): KSU, in collaboration with USDA-ARS, Cornell has developed detection and sequencing technology for ASF.
• Emerging PRRSV-2 Variant Isolation (UMN + Illinois): Minnesota and Illinois are sharing field samples to isolate and characterize emerging PRRSV-2 variants for genomic and phenotypic analysis.
• PRRSV Epitope Mapping via Immuno-Informatics (UMN): UMN applied immuno-informatic tools to systematically identify T-cell epitope content across diverse PRRSV-2 strains, advancing understanding of antigenic diversity.
• PRRS Vaccine (KSU, USDA, TSU): KSU, in collaboration with USDA-ARS and TSU, has successfully evaluated the vaccine efficacy of a prototype Interferon-Augmented PRRSV Vaccine against NADC-34 challenge, a key step in developing novel systems vaccinology approaches for swine health.
• Multivalent SIV Vaccine Challenge Study (NY + OSU): New York, in collaboration with Ohio State, will complete an immunization challenge study to assess the efficacy of a multivalent swine influenza vaccine (H1 + H3) using broadly protective platforms.
• scRNAseq and Genome Analysis (Illinois + ISU): Illinois will collaborate with Iowa State to finalize bioinformatic analysis of scRNAseq data and full-length PRRSV genome sequencing.
• PEDV/PDCoV Nanosensor Validation (Illinois + ISU): Illinois researchers will collaborate with Dr. Jianqiang Zhang (Iowa State) to validate DNA aptamer-based nanosensor tests for PEDV and PDCoV detection.
• SVA Vaccine Candidate Development (NY): New York identified a persistence marker in Senecavirus A and is leveraging this discovery to develop a non-persistent SVA vaccine candidate.
• Characterization of swine trophoblast organoids (NCSU, Duke, and Kentucky University): new model for studying vertical transmission of swine viruses (PRRSV etc).
• Validation of NanoString technology for multi-pathogen surveillance in pig production (NCSU+ ISU), facilitating the identification of pathogens involved in the porcine respiratory disease complex.
• PoAstV4 proof of primary pathogen (NCSU,UC Santa Cruz,USDA-ARS): reproduced respiratory lesions in CDCD pigs inoculated with PoAstV4 positive tissue homogenate. A shedding curve has been defined as well as the humoral immune response to infection.
• NCSU - SHIC Project Completion: Completed the epidemiological framework necessary to reconstruct vehicle movement networks and finalized the cost-benefit analysis for vehicle rerouting (2024-2025).
• NCSU Tool Deployment: Released MHASpread version 0.1.0 and provided comprehensive training to international stakeholders, achieving the milestone of equipping South American partners with U.S.-developed modeling tools to combat swine diseases like ASF.

1.   Evaluation of a Novel Swine Influenza Nanovaccine

Funding Source: USDA-AFRI (2025-39601-44637)

Amount: $649,999 | Period: 04/15/2025 – 04/14/2028

Investigators: Gourapura RJ, Kara C (OSU)

Collaborating Institutions: OH

2.   System for Early Detection of Disease in Animals

Funding Source: Kalix LLC Dayton

Amount: $35,000 | Period: 11/01/2024 – 12/15/2025

Investigators: Gourapura RJ, Kenney S (OSU)

Collaborating Institutions: OH

3.   Novel Pseudorabies Virus (PRV) Vectored Trivalent Subunit Vaccine Against Porcine Reproductive and Respiratory Syndrome, Porcine Circo, and Swine Influenza Viruses

Funding Source: USDA-AFRI (2023-07881)

Amount: $800,000 | Period: 07/01/2024 – 06/30/2028

Investigators: Chowdhury S (LSU), Gourapura RJ (OSU)

Collaborating Institutions: LA, OH

4.   Novel Broadly Protective Swine Influenza Vaccine Platforms

Funding Source: USDA-AFRI (2022-67015-36349)

Amount: $650,000 | Period: 01/01/2022 – 12/31/2025

Investigators: Diel D (Cornell), Gourapura RJ (OSU)

Collaborating Institutions: NY, OH

5.   Best Practice Considerations for PRRS Control Tools

Funding Source: AASV

Amount: $13,000 | Period: 08/01/2025 – 07/31/2026

Investigators: Arruda A (OSU)

Collaborating Institutions: OH, MN

6.   NADPRP Protecting PR Swine

Funding Source: NADPRP USDA APHIS

Amount: $221,528 | Period: 05/01/2023 – 04/30/2026

Investigators: Arruda A, Bowman A, Nolting J (OSU)

Collaborating Institutions: OH, PR

7.   Capturing Swine Movements

Funding Source: NADPRP USDA APHIS

Amount: $212,000 | Period: 11/01/2024 – 05/07/2026

Investigators: Arruda A (OSU), Cheng T-Y (MSU)

Collaborating Institutions: OH, MN

8.   Evaluating H5N1 risk to swine: mammary transmission and clinical presentation in lactating sows

Funding Source: Swine Health Information Center

Amount: $249,989 | Period: 07/01/2025 – 06/30/2026

Investigators: Warren C, Kenney S (OSU), Langel S (Case Western)

Collaborating Institutions: OH

9.   Reversion to virulence studies for African swine fever vaccines

Funding Source: USDA

Amount: $400,000 | Period: 07/01/2025 – 06/30/2026

Investigators: Kenney S, Gourapura RJ (OSU), Borca M (USDA-ARS)

Collaborating Institutions: OH, NY, KS

10. Vector competence of Texas-derived Ornithodoros turicata to transmit African swine fever virus (ASFv )

Funding Source: Department of Homeland Security

Amount: $600,000 | Period: 07/01/2025 – 06/30/2026

Investigators: Kenney S, Gourapura RJ (OSU), Saleh M, Manley H (Texas A&M)

Collaborating Institutions: OH, TX

11. Development and evaluation of a novel viral vectored vaccine to protect pigs against porcine reproductive and respiratory syndrome virus

Funding Source: Haarberg Center for Drug, Disease and Delivery (3D)

Amount: $99,597 | Period: Feb 2023 – Feb 2025

Investigators: Mor SK, Nelson E, Ly H, Sharafeldin TA, Dhanda S, Lawson S (SDSU)

Collaborating Institutions: SD

12. Surveillance, Rapid Detection, and Characterization of SARS-CoV-2 in Farmed and Wild Animals

Funding Source: USDA-APHIS

Amount: $2,156,152 | Period: Jul 2023 – Jun 2025

Investigators: Mor S, Nelson E, et al.

Collaborating Institutions: SD

13. Early detection and kinetics from oronasal exposure of 2 African Swine Fever Virus (ASFV) strains in boars and boar semen.

Funding Source: National Pork Board

Amount: $201,200 | Period: Dec 2022 – Oct 2025

Investigators: Reicks D, Ambagala A, et al.

Collaborating Institutions: SD

14. Improving molecular surveillance of PRRSV-2: Quantifying global and domestic risks of PRRSV-2 variants of concern

Funding Source: Swine Health Information Center

Amount: $65,287 | Period: 2025 – 2026

Investigators: VanderWaal K, Corzo C, et al.

Collaborating Institutions: MN

15. Rapid assessment of transmission and control of a new and rapidly spreading PRRSV variant, 1C.5.32

Funding Source: Rapid Agricultural Response Fund

Amount: $173,033 | Period: 2025 – 2027

Investigators: VanderWaal K, Corzo C, et al.

Collaborating Institutions: MN

16. Enhancing Biosecurity in Swine Operations: Investigating Wildlife Interactions on Swine Farms

Funding Source: Swine Health Information Center

Amount: $169,547 | Period: 2025 – 2027

Investigators: Paploski I, Corzo C, VanderWaal K

Collaborating Institutions: MN

17. Cooperative Agreement: Data Collection and Stakeholder Engagement for the ASF/CSF Higher Risk Surveillance Component

Funding Source: USDA-APHIS

Amount: $114,109 | Period: 2025 – 2026

Investigators: Schambow R, Perez A

Collaborating Institutions: MN

18. Veterinary Continuing Education to Enhance FAD Early Reporting and Surveillance in Non-Commercial Swine.

Funding Source: USDA NADPRP

Amount: $223,022 | Period: 2025 – 2027

Investigators: Schambow R, Perez A

Collaborating Institutions: MN

19. Real-time monitoring of performance indicators of filters and an electrostatic precipitator (ESP) to advance the control of airborne diseases in swine

Real-Time Monitoring of Filters and ESP

Funding Source: SHIC

Amount: $65,089 | Period: 11/2025 – 10/2026

Investigators: Torremorell

Collaborating Institutions: MN

20. Evaluation of cross-protective N1 swine antibodies against HPAI H5N1 clade 2.3.4.4b virus.

Funding Source: SHIC

Amount: $115,109 | Period: 04/2025 – 04/2026

Investigators: Mena J

Collaborating Institutions: MN

21. Optimizing the TELSVirus workflow for improved surveillance and characterization of swine respiratory viruses

Funding Source: SHIC

Amount: $148,107 | Period: 11/2025 – 10/2026

Investigators: Noyes N

Collaborating Institutions: MN

22. Understanding the epidemiology of influenza in growing pig herds in the U.S Midwest.

Funding Source: Merck Animal Health

Amount: $63,954 | Period: 07/2024 – 11/2025

Investigators: Torremorell

Collaborating Institutions: MN

23. Innate immune regulation and cell tropism for porcine deltacoronavirus

Funding Source: USDA Hatch

Amount: $30,892 | Period: 01/2023 – 09/2025

Investigators: Yoo D

Collaborating Institutions: IL

24. Recombination-negative, immune-enhanced, and clinically attenuated PRRSV as a vaccine platform

Funding Source: USDA NIFA

Amount: $650,000 | Period: 04/2023 – 04/2027

Investigators: Yoo D (IL), Wang Q (OH)

Collaborating Institutions: IL, OH

25. Validation of a new enrichment method for increasing sensitivity of PRRSV whole genome sequencing

Funding Source: SHIC

Amount: $98,169 | Period: 08/2024 – 07/2026

Investigators: Wang L, Yoo D

Collaborating Institutions: IL

26. A novel reverse genetics system for PRRS virus

Funding Source: USDA Hatch

Amount: $12,491 | Period: 02/2025 – 09/2026

Investigators: Yoo D

Collaborating Institutions: IL

27. Smartphone-linked field deployable test for African swine fever virus.

Funding Source: USDA Hatch

Amount: $56,738 | Period: 10/2023 – 09/2025

Investigators: Fang Y, Cunningham B, Shi J

Collaborating Institutions: IL, KS

28. Portable, rapid, sensitive, and inexpensive point-of-use biosensor technology and assays for on-farm detection of ASFV infection

Funding Source: USDA NIFA

Amount: $617,500 | Period: 09/2024 – 08/2027

Investigators: Cunningham B, Fang Y, Shi J

Collaborating Institutions: IL, KS

29. Rapid, inexpensive, simple, and sensitive on-farm detection of PEDV and PDCoV infections

Funding Source: National Pork Board

Amount: $99,415 | Period: 02/2025 – 07/2026

Investigators: Wang X, Fang Y, Zhang J

Collaborating Institutions: IL, IA

30. Molecular mechanisms of PRRSV persistence: Identification and targeting viral-host signatures for disease surveillance and prevention.

Funding Source: USDA NIFA

Amount: $635,000 | Period: 03/2023 – 02/2027

Investigators: Fang Y, Rowland R, Dekkers J, Tuggle C, Zhang J

Collaborating Institutions: IL, IA

31. A Broadly protective vaccine and novel delivery platform for influenza A virus

Funding Source: USDA NIFA

Amount: $649,896 | Period: 04/2025 – 04/2028

Investigators: Fang Y, Huber V, Tomich J

Collaborating Institutions: IL, SD, KS

32. Validation of Prognostic Biomarkers of Porcine Reproductive and Respiratory Syndrome Virus Virulence.

Funding Source: SHIC

Amount: $150,000 | Period: Dec 2024 – May 2026

Investigators: Zuckermann F, Blair B, Lowe J

Collaborating Institutions: IL, ISU

33. Fitness-virulence tradeoffs across the pathogen, pig, and population scales for genetic variants of PRRSV-2.

Funding Source: USDA

Amount: $2,999,343 (UIUC Budget: $450,000) | Period: 01/2026 – 07/2029

Investigators: VanderWaal K, Zuckermann F

Collaborating Institutions: IL, MN

34. A versatile platform for the rapid development and update of swine influenza virus vaccines

Funding Source: USDA NIFA

Amount: $649,943 | Period: TBD

Investigators: Vu H, Sillman S

Collaborating Institutions: NE

35. Development of a vaccine against highly pathogenic avian influenza viruses for use in pigs.

Funding Source: SHIC

Amount: $249,949 | Period: Jul 2025 – Dec 2026

Investigators: Vu H

Collaborating Institutions: NE, KS

36. AM02- Nipah Hendra/Nipah Vaccine for Pandemic Preparedness.

Funding Source: USDA-ARS-NACA

Amount: $387,048.51 | Period: Aug 2024 – Aug 2026

Investigators: Miller (PI), Ghmire, Kouba (KSU); Broder (USU); Korkmaz, Dhayani (UPITT); Andranov (UMD); Johnson, Hensley (USDA‑ARS‑NBAF); O’Kennedy (CSIR)Collaborating Institutions: KS, PA, MD

37. Non-Coding Regulatory RNA as Biomarkers for Epigenetic Manipulation and Antiviral

Regulation in Porcine Macrophages upon Viral Infections.

Funding Source: USDA‑NIFA‑AFRI‑CBG

Amount: $599,685 | Period: Apr 2025 – Aug 2028

Investigators: Sang, Nahashon (TSU); Miller (KSU)

Collaborating Institutions: KS, TN

38. NanoSTING PK/PD Study in the Outbred Pig Model.

Funding Source: AuraVax Therapeutics

Amount: $46,386 | Period: Jul 2025 – May 2026

Investigators: Miller, Dhakal (KSU); Cooper, Hughes, Varadarajan (AuraVax)

Collaborating Institutions: KS, TX

39. Comparative Shotgun Metatranscriptomic Analysis of Influenza A Virus Infection in Porcine Organoids.

Funding Source: Illumina (In-kind)

Amount: $20,000 | Period: 2023 – 2025

Investigators: Miller, Schirtzinger (KSU); Nelli, Gimenez‑Lirola (ISU)

Collaborating Institutions: KS, IA

40. Validation of a Live-Virus Vaccine Candidate for Efficient Attenuation and Better Protection.

Funding Source: USDA‑NIFA‑FASE‑EPSCoR

Amount: $640,000 | Period: 2020 – 2025

Investigators: Sang (TSU); Miller (KSU)

Collaborating Institutions: KS, TN

41. Intervention Strategies to Control Endemic and New Emerging and Re-Emerging Viral Diseases of Swine.

Funding Source: USDA‑NIFA‑ARS‑REE

Amount: $2.8M | Period: 2021 – 2026

Investigators: Buckley (USDA‑ARS‑NADC); Miller (KSU)

Collaborating Institutions: IA, KS

42. Partnership: Validation of Novel Transcriptomic Blood Biomarkers of Pain in Cattle and Swine after Routine Management Procedures.

Funding Source: USDA‑NIFA‑AFRI

Amount: $800,000 | Period: Apr 2022 – Mar 2026

Investigators: Coetzee, Viscardi, Weiss, Shippy, Miller (KSU); Egger, Smith, Myers, Bortoluzzi (FDA); Hutchinson, Barratt (Adelaide)

Collaborating Institutions: KS, MD

43. Development and Validation of Point of Care (POC) Assays for ASF and CSF for Rapid Outbreak Response.

Funding Source: USDA APHIS

Amount: $499,871 | Period: Mar 2021 – Feb 2024

Investigators: Pogranichniy (KSU, co‑PI); Prarat (Ohio State)

Collaborating Institutions: KS, OH

44. Symposium on International Experiences with High Consequence Animal Disease Outbreaks and Response.

Funding Source: USDA/APHIS

Amount: $40,500 | Period: May 2024 – Jan 2025

Investigators: (Not listed)

Collaborating Institutions: (Not listed)

45. 2023 International Conference of Swine Viral Diseases.

Funding Source: USDA NIFA

Amount: $25,000 | Period: Sep 2023 – Jun 2024

Investigators: R. Pogranichniy (co‑PI)

Collaborating Institutions: (Not listed)

46. Testing of Cell Lines for PRRS Virus Replication.

Funding Source: PIC North America

Amount: $68,170 | Period: Jan 2023 – Dec 2024

Investigators: R. Pogranichniy (Co‑PI)

Collaborating Institutions: (Not listed)

47. Investigating Different Methods for Decontaminating Feed Manufacturing Facilities.

Funding Source: Swine Health Information Center

Amount: $230,020 | Period: Oct 2021 – Sep 2024

Investigators: R. Pogranichniy (Co‑PI)

Collaborating Institutions: (Not listed)

48. Establishment of the Swine Trophoblast Organoid Model

Funding Source: NCSU CVM intramural grant

Amount: $20,000 | Period: July 2025 – June 2026

Investigators: Crisci E (NCSU, PI), Coyne, C. (Duke, Co-PI); Pasternak, A (KU, collaborator)

Collaborating Institutions: NCSU, Duke University, University of Kentucky

49. Predict and Protect against PRRSV: Combine PRRSV forecasting technology with vaccine efficacy prediction to prevent PRRSV outbreaks

Funding Source: USDA NIFA IDEAS (2022-68014-37266)

Amount: $1,000,000 | Period: July 2022 – June 2026

Multi-Investigators: Crisci E (NCSU PD), Machado G (NCSU, Co-PD), Kaeser T (Vienna, Co-PD)

Collaborating Institutions: NCSU, UMN, Vetmeduni Vienna, EpiVax

50. Maternal Influenza Vaccination for Generation of HA-Specific Neutralizing sIgA Antibodies in Breast Milk

Funding Source: CWRU (OH) – Gates Foundation

Amount: $225,407 | Period: October 2022 – November 2025

Investigators: Crisci E (NCSU PI, subaward)

Collaborating Institutions: NCSU, Case Western Reserve University (OH)

51. Optimizing swine sample collector network: a comprehensive analysis of sampler distribution, logistics, and national animal health laboratory network capacity

Funding Source: USDA-APHIS NADPRP

Amount: $347,062| Period: 2025-2027

Investigator: Machado G.

52. A novel multilevel model of swine disease spread to assess the effectiveness and feasibility of African swine fever control and eradication strategies

Funding Source: USDA-AFRI

Amount: $591,481| Period: 2024-2027

Investigator: Machado G.

53. Enhancing U.S. Swine Industry Readiness in Missouri By Combining Standardized On-Farm Biosecurity Plans and Animal Movement in a Data Management Tool

Funding Source: USDA-APHIS NADPRP

Amount: $271,730| Period: 2022-2025

Investigator: Machado G.

54. Descriptive Analysis of Multiple Swine Movement Networks and The Development of a Network Model to Estimate the Impacts of the Movement Restrictions Under the National African Swine Fever Response Plan.

Funding Source: USDA-APHIS NADPRP

Amount: $312,012| Period: 2022-2025

Investigador: Machado G.

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