A new indirect ELISA to identify antibodies against Senecavirus A is now available at the UMN-VDL

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Today, we are very pleased to report that a new indirect ELISA to identify Senecavirus A antibodies has been validated at the University of Minnesota and is now available for our Veterinary Diagnostic Laboratory clients. This ELISA targets specifically antibodies against Viral Protein 2 (VP2) and has a sensitivity of 94.2% and a specificity of 89.7%. The test does not cross react with antibodies against Foot-and-Mouth Disease allowing for a quick differentiation between a Senecavirus A outbreak and a costly foreign animal disease.
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Fig 1: Different ELISA types (Source: nptel.ac.in)
Abstract
Background: Senecavirus A (SVA), a member of the family Picornaviridae, genus Senecavirus, is a recently identified single-stranded RNA virus closely related to members of the Cardiovirus genus. SVA was originally identified as a cell culture contaminant and was not associated with disease until 2007 when it was first observed in pigs with Idiopathic Vesicular Disease (IVD). Vesicular disease is sporadically observed in swine, is not debilitating, but is significant due to its resemblance to foreign animal diseases, such as foot-and-mouth disease (FMD), whose presence would be economically devastating to the United States. IVD disrupts swine production until foreign animal diseases can be ruled out. Identification and characterization of SVA as a cause of IVD will help to quickly rule out infection by foreign animal diseases.
Methods: We have developed and characterized an indirect ELISA assay to specifically identify serum antibodies to SVA. Viral protein 1, 2 and 3 (VP1, VP2, VP3) were expressed, isolated, and purified from E. coli and used to coat plates for an indirect ELISA. Sera from pigs with and without IVD symptoms as well as a time course following animals from an infected farm, were analyzed to determine the antibody responses to VP1, VP2, and VP3.
Results: Antibody responses to VP2 were higher than VP1 and VP3 and showed high affinity binding on an avidity ELISA. ROC analysis of the SVA VP2 ELISA showed a sensitivity of 94.2% and a specificity of 89.7%. Compared to IFA, the quantitative ELISA showed an 89% agreement in negative samples and positive samples from 4–60 days after appearance of clinical signs. Immune sera positive for FMDV, encephalomyocarditis virus, and porcine epidemic diarrhea virus antibodies did not cross-react.
Conclusions: A simple ELISA based on detection of antibodies to SVA VP2 will help to differentially diagnose IVD due to SVA and rule out the presence of economically devastating foreign animal diseases.

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Development and validation of a competitive ELISA as a screening test for Senecavirus A

An article published in the Journal of Veterinary Diagnostic Investigation (JVDI) presents a competitive Enzyme-Linked ImmunoSorbent Assay (cELISA) and a virus neutralization test (VNT), both validated for the screening of Senecavirus A in a research setting, by the National Centre for Foreign Animal disease (NCFAD). The diagnostic specificity and sensitivity were 98.2% and 96.9% for the cELISA, and 99.6% (99.0–99.9%) and 98.2% (95.8–99.4%) for the VNT, respectively.

In Canada and the USA alike, Senecavirus A is a challenge for producers and veterinarians because of its clinical similarity to Food and Mouth Disease (FMD). Indeed, Senecavirus A, is a causative agent of swine vesicular disease with lesions developing on the snout, around the mouth and on the coronary band of the feet. Therefore, being able to differentiate Senecavirus A infections from FMD rapidly is of utmost importance to be able to take the appropriate measures.

The University of Minnesota, Veterinary Diagnostic Laboratory has developed an ImmunoFluorescence Assay (IFA) to detect antibodies against Senecavirus A. This test was used as a reference for the validation of the cELISA and VNT established by Drs. Goolia, Yang, Babiuk, and Nfon from NCFAD in collaboration with Drs. Vannucci and Patnayak from the UMN-VDL.

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Abstract: Senecavirus A (SVA; family Picornaviridae) is a nonenveloped, single-stranded RNA virus associated with idiopathic vesicular disease (IVD) in swine. SVA was detected in pigs with IVD in Brazil, United States, Canada, and China in 2015, triggering the need to develop and/or validate serologic assays for SVA. Our objective was to fully validate a previously developed competitive enzyme-linked immunosorbent assay (cELISA) as a screening test for antibodies to SVA. Additional objectives included the development and validation of a virus neutralization test (VNT) as a confirmatory test for SVA antibody detection, and the comparison of the cELISA, VNT, and an existing immunofluorescent antibody test (IFAT) for the detection of SVA antibodies in serial bleeds from SVA outbreaks. The diagnostic specificity and sensitivity were 98.2% (97.2–98.9%) and 96.9% (94.5–98.4%) for the cELISA, and 99.6% (99.0–99.9%) and 98.2% (95.8–99.4%) for the VNT, respectively. There was strong agreement among cELISA, VNT, and IFAT when compared based on kappa coefficient. Based on these performance characteristics, these tests are considered suitable for serologic detection of SVA in pigs.

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Using Machine Learning to Predict Swine Movements

A collaborative work between the University of Minnesota, UC- Davis, and Pipestone Veterinary Services was published this past month in the journal Frontiers in Veterinary Science.
Between-farm animal movement, despite being an essential factor of infectious disease spread is not currently recorded in the US. The objective of this project was to create a model to predict animal movement based on between-site distance, ownership, and production type of the sending and receiving farms. The model was able to predict animal movement in the south-central region of the study area with a high aggregation. It also showed an overlap with the distribution of outbreaks of porcine reproductive and respiratory syndrome (PRRS) in this area.

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Abstract: Between-farm animal movement is one of the most important factors influencing the spread of infectious diseases in food animals, including in the US swine industry. Understanding the structural network of contacts in a food animal industry is prerequisite to planning for efficient production strategies and for effective disease control measures. Unfortunately, data regarding between-farm animal movements in the US are not systematically collected and thus, such information is often unavailable. In this paper, we develop a procedure to replicate the structure of a network, making use of partial data available, and subsequently use the model developed to predict animal movements among sites in 34 Minnesota counties. First, we summarized two networks of swine producing facilities in Minnesota, then we used a machine learning technique referred to as random forest, an ensemble of independent classification trees, to estimate the probability of pig movements between farms and/or markets sites located in two counties in Minnesota. The model was calibrated and tested by comparing predicted data and observed data in those two counties for which data were available. Finally, the model was used to predict animal movements in sites located across 34 Minnesota counties. Variables that were important in predicting pig movements included between-site distance, ownership, and production type of the sending and receiving farms and/or markets. Using a weighted-kernel approach to describe spatial variation in the centrality measures of the predicted network, we showed that the south-central region of the study area exhibited high aggregation of predicted pig movements. Our results show an overlap with the distribution of outbreaks of porcine reproductive and respiratory syndrome, which is believed to be transmitted, at least in part, though animal movements. While the correspondence of movements and disease is not a causal test, it suggests that the predicted network may approximate actual movements. Accordingly, the predictions provided here might help to design and implement control strategies in the region. Additionally, the methodology here may be used to estimate contact networks for other livestock systems when only incomplete information regarding animal movements is available.

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Reducing the likelihood of a piglet reservoir when dealing with influenza in swine herds

Drs. White, Torremorell and Craft from the University of Minnesota recently published an article in Preventative Veterinary Medicine regarding practices that can decrease the likelihood of creating an endemic piglet reservoir in the case of an infection by swine influenza. Indeed, a stochastic model was developed considering that the pigs were in one of the following categories: Susceptible, Exposed,  Infectious, Recovered, or Vaccinated. Loss of immunity over time and differences between naturally infected and vaccinated animals were taken into account. Several scenarios were evaluated regarding their impact on piglet prevalence: timing of gilt introductions, gilt separation, gilt vaccination upon arrival, early weaning, and sow vaccination strategies.

In this model, homologous mass vaccination and early weaning were the most efficacious interventions. By combining frequent homologous mass vaccination, early weaning, gilt separation, gilt vaccination and longer periods between gilt introductions reduced endemic prevalence overall by 51% relative to the null scenario and the endemic prevalence in piglets by 74%.

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Abstract: Recent modelling and empirical work on influenza A virus (IAV) suggests that piglets play an important role as an endemic reservoir. The objective of this study is to test intervention strategies aimed at reducing the incidence of IAV in piglets and ideally, preventing piglets from becoming exposed in the first place. These interventions include biosecurity measures, vaccination, and management options that swine producers may employ individually or jointly to control IAV in their herds. We have developed a stochastic Susceptible-Exposed-Infectious-Recovered-Vaccinated (SEIRV) model that reflects the spatial organization of a standard breeding herd and accounts for the different production classes of pigs therein. Notably, this model allows for loss of immunity for vaccinated and recovered animals, and for vaccinated animals to have different latency and infectious periods from unvaccinated animals as suggested by the literature. The interventions tested include: (1) varied timing of gilt introductions to the breeding herd, (2) gilt separation (no indirect transmission to or from the gilt development unit), (3) gilt vaccination upon arrival to the farm, (4) early weaning, and (5) vaccination strategies of sows with different timing (mass and pre-farrow) and efficacy (homologous vs. heterologous). We conducted a Latin Hypercube Sampling and Partial Rank Correlation Coefficient (LHS-PRCC) analysis combined with random forest analysis to assess the relative importance of each epidemiological parameter in determining epidemic outcomes. In concert, mass vaccination, early weaning of piglets (removal 0–7 days after birth), gilt separation, gilt vaccination, and longer periods between introductions of gilts (6 months) were the most effective at reducing prevalence. Endemic prevalence overall was reduced by 51% relative to the null case; endemic prevalence in piglets was reduced by 74%; and IAV was eliminated completely from the herd in 23% of all simulations. Importantly, elimination of IAV was most likely to occur within the first few days of an epidemic. The latency period, infectious period, duration of immunity, and transmission rate for piglets with maternal immunity had the highest correlation with three separate measures of IAV prevalence; therefore, these are parameters that warrant increased attention for obtaining empirical estimates. Our findings support other studies suggesting that piglets play a key role in maintaining IAV in breeding herds. We recommend biosecurity measures in combination with targeted homologous vaccination or vaccines that provide wider cross-protective immunity to prevent incursions of virus to the farm and subsequent establishment of an infected piglet reservoir.

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Advances in Mycoplasma hyopneumoniae elimination: a podcast series

This past month, the Morrison group invited Dr. Paul Yeske, swine practitioner at the Swine Vet Center (St. Peter, MN), Dr. Amanda Sponheim, PhD candidate at the University of Minnesota and Support Veterinarian at Boerhinger Ingelheim, and Dr. Maria Pieters from the University of Minnesota to discuss the latest progress made in successfully eliminating Mycoplasma hyopeumoniae from swine herds. Dr. Pieters is the head of the MycoLab at the College of Veterinary Medicine and focuses on diagnostics and epidemiology of swine mycoplasms to help veterinarians control associated diseases.

  1. History of Mycoplasma hyopneumoniae herd elimination and practices: podcast
  2. Sampling techniques and protocols to use during the process of elimination: podcast
  3. Starting the elimination: when is day zero? podcast

The podcasts in the press