Veterinary Medicine Commons^™

Comparison Of Two Diagnostic Assays For The Detection Of Serum Neutralizing Antibody To Porcine Epidemic Diarrhea Virus, Justin Brown, Korakrit Poonsuk, Ting-Yu Cheng, Chris Rademacher, Erin Kalkwarf, Liying Tian, Lauren A. Mckeen, Chong Wang, Luis Gimenez-Lirola, David Baum, Locke A. Karriker

School of Veterinary and Biomedical Sciences: Faculty Publications

Lactogenic immunity is important for the protection of piglets against many pathogens including porcine epidemic diarrhea virus. Circulating neutralizing antibodies levels in sow sera may help determine if a detectable immune response could confer protection to piglets. Neutralizing antibodies can be detected through various diagnostic assays. This study evaluated the diagnostic characteristics of two neutralizing antibody assays for porcine epidemic diarrhea virus neutralizing antibodies in serum of challenged gilts. Four treatment groups, control, non-vaccinated, vaccinated prior to challenge, and vaccinated following challenge, were comprised of 20 gilts. Serum sample were collected from each gilt prior to and following challenge with …

Comparison Of Two Diagnostic Assays For The Detection Of Serum Neutralizing Antibody To Porcine Epidemic Diarrhea Virus, Justin Brown, Korakrit Poonsuk, Ting-Yu Cheng, Chris Rademacher, Erin Kalkwarf, Liying Tian, Lauren A. Mckeen, Chong Wang, Luis Gimenez-Lirola, David Baum, Locke A. Karriker

School of Veterinary and Biomedical Sciences: Faculty Publications

Lactogenic immunity is important for the protection of piglets against many pathogens including porcine epidemic diarrhea virus. Circulating neutralizing antibodies levels in sow sera may help determine if a detectable immune response could confer protection to piglets. Neutralizing antibodies can be detected through various diagnostic assays. This study evaluated the diagnostic characteristics of two neutralizing antibody assays for porcine epidemic diarrhea virus neutralizing antibodies in serum of challenged gilts. Four treatment groups, control, non-vaccinated, vaccinated prior to challenge, and vaccinated following challenge, were comprised of 20 gilts. Serum sample were collected from each gilt prior to and following challenge with …

Detection And Phylogenetic Analysis Of Porcine Epidemic Diarrhea Virus Based On The Orf3 Gene In Pigs In The North Of Vietnam, Tran Duc Hoan, Doan Thi Thao, Nguyen Thi Huong Giang, Nguyen Thi Thu Huyen, Nguyen Thi Thu Huyen

The Thai Journal of Veterinary Medicine

To identify further the infectivity, genetic diversity and molecular characteristics of the porcine epidemic diarrhea virus (PEDV). 286 samples of the intestinal tissue of pigs with severe diarrhea were assessed from 5 provinces in the north of Vietnam. The full-length ORF3 gene of 20 representative field strains from 36 farms in 5 provinces was sequenced and analyzed. The prevalence of PEDV was 90.56% and was detected in 259 of 286 samples, with 94.44% (34 of 36) of pig farms harboring the disease. Based on the phylogenetic analysis of the ORF3 genes, our isolates all fell into group G2 (variants) and …

Expression Of Recombinant S2 Domain Of Spike Protein Of Porcine Epidemic Diarrhea Virus, Jiraporn Sritun, Natnaree Inthong, Siriluk Jala, Sakuna Phatthanakunanan, Kaitkanoke Sirinarumitr, Preeda Lertwatcharasarakul, Theerapol Sirinarumitr

The Thai Journal of Veterinary Medicine

Porcine epidemic diarrhea virus (PEDV) is the causative agent of severe watery diarrhea in suckling piglets. PEDV outbreaks have been consistently reported in Europe, Asian countries and Thailand. The recombinant protein, especially the spike (S) protein, from field strain PEDV could be used to develop a sensitive diagnostic test or an effective vaccine. In this study, the recombinant S2 domain protein from the PEDV Thai isolate was expressed using the E. coli expression system. The full length S2 domain of the S gene of the PEDV G2b Thai isolate was amplified using a PCR technique and was cloned into the …

Piglet Immunization With A Spike Subunit Vaccine Enhances Disease By Porcine Epidemic Diarrhea Virus, Jieshi Yu, Chithra Sreenivasan, Tirth Uprety, Rongyuan Gao, Chen Huang, Ella J. Lee, Steven Lawson, Julie Nelson, Jane Christopher-Hennings, Radhey S. Kaushik, Eric Nelson, Diego G. Diel, Ben M. Hause, Feng Li, Dan Wang

Maxwell H. Gluck Equine Research Center Faculty Publications

Immunization with an insect cell lysate/baculovirus mixture containing recombinant porcine epidemic diarrhea virus (PEDV) spike protein induced high levels of neutralizing antibodies in both mice and piglets. However, immunization of piglets with this vaccine resulted in enhancement of disease symptoms and virus replication in vaccine recipients exposed to PEDV challenge. Thus, these observations demonstrate a previously unrecognized challenge of PEDV vaccine research, which has important implications for coronavirus vaccine development.

Quantification Of Semi-Truck Cab Decontamination, C. Grace Elijah, Cassandra K. Jones, Caitlin Evans, Haley K. Wecker, Charles R. Stark, Jianfa Bai, Elizabeth G. Poulsen-Porter, Allison K. Blomme, Jason C. Woodworth, Chad B. Paulk, Jordan T. Gebhardt

Kansas Agricultural Experiment Station Research Reports

Evidence suggests that the inside of vehicle cabs used for feed delivery may serve as a potential source for disease, yet there are no standardized protocols or scientific evidence for methods of their disinfection. Therefore, the objective of this project was to evaluate commercially available disinfectants and disinfection application methods against PEDV and PRRSV on various surfaces within semi-truck cabs. Three different surface types common in vehicle cabs (fabric, plastic, and rubber) were cut into 4 × 4 inch coupons and inoculated with either PEDV or PRRSV. Once inoculated, surfaces were placed in one of 3 semi-truck cabs and the …

Effects Of Medium Chain Fatty Acid Application In Swine Feed On Porcine Epidemic Diarrhea Virus, A. B. Lerner, R. A. Cochrane, J. T. Gebhardt, S. S. Dritz, C. K. Jones, J. M. Derouchey, M. D. Tokach, R. D. Goodband, J. Bai, E. Porter, J. Anderson, P. C. Gauger, D. R. Magstadt, J. Zhang, B. Bass, T. P. Karnezos, B. De Rodas, J. C. Woodworth

Kansas Agricultural Experiment Station Research Reports

Medium chain fatty acid (MCFA) application has been identified as a promising strategy to decrease viral pathogen transmission in swine feed. Four experiments were conducted to: 1) determine if MCFAs are effective when applied to feed both prior to and after porcine epidemic diarrhea virus (PEDV) inoculation measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR), 2) evaluate the effects of varying amounts and combinations of MCFA measured by qRT-PCR, and 3) evaluate selected MCFA treatments in a bioassay. In Exp. 1, treatments were arranged in a 2 × 2 + 1 factorial with the main effects of chemical treatment …

Quantifying Medium Chain Fatty Acid Mitigation Activity Over Time Against Porcine Epidemic Diarrhea Virus In Nursery Pig Diets, J. T. Gebhardt, J. C. Woodworth, M. D. Tokach, J. M. Derouchey, R. D. Goodband, C. K. Jones, S. S. Dritz

Kansas Agricultural Experiment Station Research Reports

Medium chain fatty acids (MCFA) are six to twelve carbon length molecules that have shown significant promise as potential mitigants of biological hazards in feed and feed ingredients. The use of residual duration of activity approaches, such as MCFA, have significant advantages compared to point-in-time mitigation strategies. The primary advantage of MCFA is the ability to mitigate the risks generated by post-processing contamination; however, the duration of mitigation activity has not been established. Therefore, the objective of this experiment was to characterize the mitigation properties of MCFA-treated swine feed 40 d following feed manufacturing. Treatments (n = 8) consisted of …

Evaluating The Inclusion Level Of Medium Chain Fatty Acids To Reduce The Risk Of Porcine Epidemic Diarrhea Virus In Complete Feed And Spray-Dried Animal Plasma, R. A. Cochrane, S. S. Dritz, J. C. Woodworth, A. R. Huss, C. R. Stark, M. Saensukjaroenphon, J. M. Derouchey, M. D. Tokach, R. D. Goodband, J. Bai, Qi Chen, Jianqiang Zhang, Phillip Charles Gauger, Rachel J. Derscheid, Drew Robert Magstadt, Paulo Elias Arruda, Alejandro Ramirez, Rodger G. Main, C. K. Jones

Kansas Agricultural Experiment Station Research Reports

Research has confirmed that chemical treatments, such as medium chain fatty acids (MCFA) and commercial formaldehyde, can be effective to reduce the risk of porcine epidemic diarrhea virus (PEDV) cross-contamination in feed. However, the efficacy of MCFA levels below 2% inclusion is unknown. The objective of this experiment was to evaluate if a 1% inclusion of MCFA is as effective at PEDV mitigation as a 2% inclusion or formaldehyde in swine feed and spray-dried animal plasma (SDAP). Treatments were arranged in a 4 × 2 × 7 plus 2 factorial with 4 chemical treatments: 1) PEDV positive with no chemical …

Porcine Epidemic Diarrhea Virus Surface Decontamination Strategies Using Chemical Sanitizing To Reduce The Quantity Of Pedv Rna On Feed Manufacturing Surfaces With Environmental Swabbing, M. Muckey, S. S. Dritz, J. C. Woodworth, C. R. Stark, J. Bai, Jianqiang Zhang, Phillip Charles Gauger, Rodger G. Main, C. K. Jones

Kansas Agricultural Experiment Station Research Reports

Porcine Epidemic Diarrhea virus (PEDV) is a possible hazard in feed mills that could impact pig health. If the virus enters a feed mill, it quickly becomes widely distributed and is difficult to decontaminate from surfaces.6,7 The objective of this study was to evaluate a variety of liquid and dry chemical treatments that could be used as sanitizers to reduce the amount of PEDV found on feed manufacturing surfaces in mills. This experiment was replicated 3 times and was designed in a 5 × 10 factorial with main effects of 5 different feed manufacturing surfaces and 10 sanitizing treatments. Surfaces …

Evaluating The Impact Of Vevovitall And/Or Crina As Potential Porcine Epidemic Diarrhea Virus Mitigation Strategies As Determined By Polymerase Chain Reaction Analysis And Bioassay, J. T. Gebhardt, J. C. Woodworth, C. K. Jones, M. D. Tokach, J. M. Derouchey, R. D. Goodband, R. A. Cochrane, C. R. Stark, J. Bergstrom, Phillip Charles Gauger, J. Bai, Qi Chen, Jianqiang Zhang, Rodger G. Main, S. S. Dritz

Kansas Agricultural Experiment Station Research Reports

Feed and feed ingredients have been shown to be potential vectors of porcine epidemic diarrhea virus (PEDV). Potential strategies to mitigate the risk of disease transmission via feed and feed ingredients would be valuable to the swine and feed milling industries. Therefore, the objective of this experiment was to determine the impact of VevoVitall (5,000 ppm; DSM Nutritional Products Inc., Parsipanny, NJ), CRINA (200 ppm; DSM Nutritional Products Inc., Parsipanny, NJ), and a combination of both products (COMBINATION; 5,000 ppm VevoVitall and 200 ppm CRINA) as feed additives with potential to mitigate the risk of PEDV, in swine gestation diet …

Development And Characterization Of A Recombinant Orf Virus Vector Expressing The Spike Protein Of Porcine Epidemic Diarrhea Virus, Kyle Hain

Electronic Theses and Dissertations

Orf virus (ORFV), the type member of the genus Parapoxvirus of the family Poxviridae, causes orf or cutaneous pustular dermatitis in sheep and goats. ORFV is a ubiquitous virus capable of re-infecting its hosts multiple times over time. ORFV causes a non-systemic, self-limiting disease which is usually restricted to the skin surrounding the virus entry sites. ORFV has evolved several immunomodulatory proteins (IMPs) that evade and/or modulate host immune responses to infection and contribute to virus virulence and disease pathogenesis. Given biological properties and unique immunomodula tor y properties, ORFV has gained significant attention in recent years for its …

Assessing The Effects Of Medium Chain Fatty Acids And Fat Sources On Porcine Epidemic Diarrhea Virus Viral Rna Stability And Infectivity, R. A. Cochrane, S. S. Dritz, J. C. Woodworth, A. R. Huss, C. R. Stark, M. Saensukjaroenphon, J. M. Derouchey, M. D. Tokach, R. D. Goodband, J. Bai, Qi Chen, Jianqiang Zhang, Phillip Charles Gauger, Rachel J. Derscheid, Rodger G. Main, C. K. Jones

Kansas Agricultural Experiment Station Research Reports

Research has confirmed that chemical treatments, such as medium chain fatty acids (MCFA) and commercial formaldehyde, can be effective to reduce the risk of porcine epidemic diarrhea virus (PEDV) cross-contamination in feed. However, the efficacy of individual MCFA levels are unknown. The objective of this study is to compare the efficacy of commercially-available sources of MCFA and other fat sources versus a synthetic custom blend of MCFA to minimize the risk of PEDV cross-contamination as measured by qRT-PCR and bioassay. Treatments were arranged in a 17 × 4 plus 1 factorial with 17 chemical treatments: 1) Positive control with PEDV …

Evaluation Of The Effects Of Flushing Feed Manufacturing Equipment With Chemically- Treated Rice Hulls On Porcine Epidemic Diarrhea Virus Cross Contamination During Feed Manufacturing, J. T. Gebhardt, J. C. Woodworth, C. K. Jones, Phillip Charles Gauger, M. D. Tokach, J. M. Derouchey, R. D. Goodband, M. Muckey, R. A. Cochrane, M. Niederwerder, C. R. Stark, J. Bai, Qi Chen, Jianqiang Zhang, Alejandro Ramirez, Rachel J. Derscheid, Rodger G. Main, S. S. Dritz

Kansas Agricultural Experiment Station Research Reports

Various strategies have been proposed to mitigate potential risk of porcine epidemic diarrhea virus (PEDV) transmission via feed and feed ingredients. Wet decontamination has been found to be the most effective decontamination of feed mill surfaces; however, this is not practical on a commercial feed production-scale. Another potential mitigation strategy, easier to implement, would be using chemically-treated rice hulls flushed through the feed manufacturing equipment. The objective of this experiment was to determine the impact of MCFA- or formaldehyde-treated rice hull flush batches as potential PEDV mitigation strategies during feed manufacturing. Feed without evidence of PEDV RNA contamination was inoculated …

An Inactivated Vaccine Made From A U.S. Field Isolate Of Porcine Epidemic Disease Virus Is Immunogenic In Pigs As Demonstrated By A Dose-Titration, Emily A. Collin, Srivishnupriya Anbalagan, Faten Okda, Ron Bateman, Eric A. Nelson, Ben M. Hause

Veterinary and Biomedical Sciences Faculty Publications

Background: Porcine epidemic diarrhea virus (PEDV), a highly pathogenic and transmissible virus in swine, was first detected in the U.S. in May, 2013, and has caused tremendous losses to the swine industry. Due to the difficulty in isolating and growing this virus in cell culture, few vaccine studies using cell culture propagated PEDV have been performed on U.S. strains in pigs. Therefore, the objective of this study was to evaluate the humoral immune response to the selected inactivated PEDV vaccine candidate in a dose-titration manner.
Results: PEDV was isolated from a pig with diarrhea and complete genome sequencing found >99% …

Effect Of Thermal Mitigation On Porcine Epidemic Diarrhea Virus (Pedv)- Contaminated Feed, R. A. Cochrane, L. L. Schumacher, S. S. Dritz, J. C. Woodworth, A. R. Huss, C. R. Stark, J. M. Derouchey, M. D. Tokach, R. D. Goodband, J. Bai, Qi Chen, Jianqiang Zhang, Phillip Charles Gauger, Rodger G. Main, C. K. Jones

Kansas Agricultural Experiment Station Research Reports

Porcine Epidemic Diarrhea Virus (PEDV) is primarily transmitted by fecal-oral contamination. However, epidemiological evidence has shown that swine feed and ingredients may serve as potential vectors of transmission. Since it is known that PEDV is a heat-sensitive virus, we hypothesized that a conditioner and pellet mill mimicking commercial thermal processing would mitigate PEDV infectivity. To test this hypothesis, two experiments were designed to determine if different pellet mill conditioner retention times or temperatures would impact PEDV infectivity determined by polymerase chain reaction (PCR) analysis and bioassay. For the first study, a 3×3×2 factorial was utilized, with three pelleting temperatures (155, …

Evaluating The Effect Of Manufacturing Porcine Epidemic Diarrhea Virus (Pedv)-Contaminated Feed On Subsequent Feed Mill Environmental Surface Contamination, L. L. Schumacher, R. A. Cochrane, C. E. Evans, J. R. Kalivoda, J. C. Woodworth, C. R. Stark, C. K. Jones, Rodger G. Main, Jianqiang Zhang, S. S. Dritz, Phillip Charles Gauger

Kansas Agricultural Experiment Station Research Reports

This study aimed to utilize the only known pilot feed mill facility approved for pathogenic feed agent use in the United States to evaluate the effect of manufacturing Porcine Epidemic Diarrhea Virus (PEDV)-contaminated feed on subsequent feed mill environmental surface contamination. In this study, PEDV inoculated feed was manufactured and conveyed on equipment along with four subsequent batches of PEDV-free feed. Equipment and environmental surfaces were sampled using swabs and analyzed for the presence of PEDV RNA by PCR. The experiment was replicated three times with decontamination of the feed mill and all equipment between replications. Overall, environmental swabs indicated …

Determining The Minimum Infectious Dose Of Porcine Epidemic Diarrhea Virus (Pedv) In A Feed Matrix, L. L. Schumacher, J. C. Woodworth, C. R. Stark, C. K. Jones, R. A. Hesse, Rodger G. Main, Jianqiang Zhang, Phillip Charles Gauger, S. S. Dritz, M. D. Tokach

Kansas Agricultural Experiment Station Research Reports

Understanding the magnitude of transmissible risk Porcine Epidemic Diarrhea Virus (PEDV)-infected feed imposes and establishing the minimum infectious dose of PEDV in a feed matrix are important components in strengthening virus prevention and control methods. In this study, an experiment was performed involving 30 crossbred, 10-d-old pigs that were used as a bioassay model for the minimum infectious dose of PEDV in feed. The PEDV was first diluted using tissue culture media to form 8 serial 10-fold dilutions. An aliquot of the original stock virus at 5.6 x 10⁵tissue culture infectious dose/ml (TCID₅₀/ml), each serial PEDV …

Utilizing Feed Sequencing To Decrease The Risk Of Porcine Epidemic Diarrhea Virus (Pedv) Cross-Contamination During Feed Manufacturing, L. L. Schumacher, R. A. Cochrane, J. C. Woodworth, C. R. Stark, C. K. Jones, Rodger G. Main, Jianqiang Zhang, Phillip Charles Gauger, S. S. Dritz, M. D. Tokach

Kansas Agricultural Experiment Station Research Reports

Understanding key points of potential cross-contamination during the feed manufacturing process is important to developing efficacious methods to control or prevent transmission of pathogens into swine diets. In this study, an experiment was conducted involving 30 crossbred 10-d-old pigs that were used as a bioassay model for Porcine Epidemic Diarrhea Virus (PEDV) to determine the effects of feed batch sequencing on PEDV cross-contamination and subsequent infectivity. PEDV with a PCR cycle threshold value (Ct) of 11 was uniformly mixed into 4.5 kg of swine diet using a stainless steel bench top mixer validated for mixing efficiency. The inoculated feed was …

Evaluating Chemical Mitigation Of Porcine Epidemic Diarrhea Virus (Pedv) In Swine Feed And Ingredients, R. A. Cochrane, S. S. Dritz, J. C. Woodworth, A. R. Huss, C. R. Stark, R. A. Hesse, Jianqiang Zhang, M. D. Tokach, J. Bai, C. K. Jones

Kansas Agricultural Experiment Station Research Reports

Porcine Epidemic Diarrhea Virus (PEDV) is primarily transmitted by fecal-oral contamination. Research has confirmed swine feed or ingredients as potential vectors of transmission, so strategies are needed to mitigate PEDV in feed. The objective of this experiment was to evaluate the effectiveness of various chemical additives to prevent or mitigate post-processing PEDV contamination in swine feed and ingredients. Treatments were arranged in a 7 × 4 factorial with seven chemical treatments and four feed matrices. The chemical treatments included: negative control with no chemical addition, 0.3% commercial formaldehyde product, 1% sodium bisulfate, 1% sodium chlorate, 3% custom organic acid blend …

An Evaluation Of A Liquid Antimicrobial (Sal Curb®) For Reducing The Risk Of Porcine Epidemic Diarrhea Virus Infection Of Naïve Pigs During Consumption Of Contaminated Feed, Scott Dee, Casey Neill, Travis Clement, Jane Christopher-Hennings, Eric A. Nelson

Veterinary and Biomedical Sciences Faculty Publications

Background: Since its initial detection in May 2013, porcine epidemic diarrhea virus (PEDV) has spread rapidly throughout the US swine industry. Recently, contaminated feed was confirmed as a vehicle for PEDV infection of naïve piglets. This research provides in vivo data supporting the ability of a liquid antimicrobial product to reduce this risk.
Results: Sal CURB® (Kemin Industries, Des Moines, IA, USA) is a FDA-approved liquid antimicrobial used to control Salmonella contamination in poultry and swine diets. To test its effect against PEDV, Sal CURB®-treated feed was spiked with a stock isolate of PEDV (Ct = 25.22), which PEDV-naïve piglets …

An Evaluation Of Contaminated Complete Feed As A Vehicle For Porcine Epidemic Diarrhea Virus Infection Of Naïve Pigs Following Consumption Via Natural Feeding Behavior: Proof Of Concept, Scott Dee, Travis Clement, Adam Schelkopf, Joel Nerem, David Knudsen, Jane Christopher-Hennings, Eric A. Nelson

Veterinary and Biomedical Sciences Faculty Publications

Background: Since its initial detection in May 2013, porcine epidemic diarrhea virus (PEDV) has spread rapidly throughout the US swine industry. Initially, contaminated feed was proposed as a risk factor for PEDV; however, data were not available to support this theory. Here we provide proof of concept of this risk by describing a novel means for recovering PEDV-contaminated complete feed material from commercial swine sites and conducting an in vivo experiment to prove its infectivity.

Results: For on-farm detection of PEDV RNA in feed, paint rollers were used to collect material from at-risk feed bins from 3 clinically affected breeding …