Pathogenesis of porcine epidemic diarrhea virus isolate (US/Iowa/18984/2013) in 3-week-old weaned pigs

Madson, Darin M.; Magstadt, Drew R.; Arruda, Paulo; Hoang, Hai; Sun, Dong; Bower, Leslie; Bhandari, Mahesh; Burrough, Eric; Gauger, Phillip C.; Pillatzki, Angela; Stevenson, Gregory W.; Wilberts, Bailey L.; Brodie, Joseph; Harmon, Karen M.; Wang, Chong; Main, Rodger; Zhang, J.; Yoon, Kyoung‐Jin

doi:10.1016/j.vetmic.2014.09.002

Cited by 160 publications

(209 citation statements)

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“…19 PEDV replication in mature enterocytes is rapid, with previous investigations demonstrating antigen or detecting nucleic acid 24 hpi. 2,7,11 In the current study, PEDV antigen and RNA were detected at 12 hpi. It is possible that PEDV RNA detected by fecal swab collection at 12 hpi represented the inoculum as it passed through the intestine.…”

Section: Discussionmentioning

confidence: 51%

“…Sections of fixed small intestine for microscopic evaluation included standardized locations of duodenum, proximal jejunum, midjejunum, distal jejunum, and ileum. 11 Real-time Reverse Transcription PCR Swabs, tissue homogenates, and sera were processed on the day of collection. Briefly, fecal swabs were added to tubes containing 1 ml of 0.01M phosphate-buffered saline (pH 7.4) without calcium and magnesium (GIBCO/Life Technologies, Carlsbad, CA, USA) and vortexed.…”

Section: Sample Collection and Clinical Observationsmentioning

confidence: 99%

“…All samples were stored at -80 C until viral RNA was extracted with the MagMAX Viral RNA Isolation Kit (Life Technologies, Carlsbad, CA, USA) using a modified high-volume extraction protocol per manufacturer instructions. Reverse transcription quantitative PCR was performed as previously described 11 alongside viral standards with known infectivity titers (PFU) for quantification. Each cycle threshold value was then converted to viral titer, PFU equivalent per milliliter, estimated from a standard curve generated from the viral standards of known titers.…”

Section: Sample Collection and Clinical Observationsmentioning

confidence: 99%

“…Mean villus length and crypt depth were used for statistical analysis and for determining villus height:crypt depth ratios. 11 Immunohistochemical staining for PEDV antigen was performed on duodenum, proximal jejunum, midjejunum, distal jejunum, and ileum sections from all pigs. Antigen labeling in each section was semiquantitatively assessed on a scale ranging from 0 to 3, where 0 signifies no signaling detected and 3 indicates that >50% of villous enterocytes were observed with positive staining.…”

Section: Histology Intestinal Morphometry and Immunohistochemistrymentioning

confidence: 99%

“…Antigen labeling in each section was semiquantitatively assessed on a scale ranging from 0 to 3, where 0 signifies no signaling detected and 3 indicates that >50% of villous enterocytes were observed with positive staining. 11 All scores were recorded by a single pathologist blinded to individual piglet identification and treatment group. In addition, mesenteric lymph node, colon, spleen, stomach, heart, spleen, liver, and kidney were evaluated for PEDV antigen by immunohistochemistry (IHC).…”

Section: Histology Intestinal Morphometry and Immunohistochemistrymentioning

confidence: 99%

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Characterization of Porcine Epidemic Diarrhea Virus Isolate US/Iowa/18984/2013 Infection in 1-Day-Old Cesarean-Derived Colostrum-Deprived Piglets

et al. 2015

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Porcine epidemic diarrhea virus (PEDV) was first recognized in North America in April 2013 and has since caused devastating disease. The objective of this study was to characterize disease and viral detection associated with an original North American PEDV isolate inoculated in neonatal piglets. Thirty-six 1-day-old cesarean-derived and colostrum-deprived piglets were randomly assigned to the control (n ¼ 16) or challenged group (n ¼ 20); the latter were orogastrically inoculated with 1 ml of US/Iowa/ 18984/2013 PEDV isolate titered at 1 Â 10 3 plaque-forming units per milliliter. Rectal swabs were collected from all piglets prior to inoculation and every 12 hours postinoculation (hpi) thereafter, with 4 control and 5 challenged piglets euthanized at 12, 24, 48, and 72 hpi. One piglet had a positive real-time quantitative polymerase chain reaction test on rectal swab at 12 hpi, and all remaining piglets were positive thereafter, with highest viral quantities detected at 24 and 36 hpi. Diarrhea was evident in 30% and 100% of challenged piglets at 18 and 24 hpi, respectively. Viral antigen was detected in enterocytes by immunohistochemistry in the duodenum and ileum of piglets euthanized at 12 hpi and was apparent throughout the small intestine of all piglets thereafter, with villus height:crypt depth ratios consistently below 4:1. Viremia was confirmed in 18 of 20 pigs at euthanasia. Clinical disease was severe and developed rapidly following infection with an original North American PEDV isolate, with lesions, viremia, and antigen detection possible by 12 hpi.

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