dr. dick hesse - recent emergence of swine coronaviruses in north america

2014 Allen D. Leman Swine Conference

St. Paul River Centre, St. Paul, Minnesota

Science-driven solutionsTM 

Carlos Pijoan Lecture

Recent Emergence of Swine Coronaviruses:

A Virologist’s Perspective

Dr. Carlos Pijoan

A man of many talentsA man who cared

An inspiration to all of us

Recent Emergence of Swine Coronaviruses:

A Virologist’s Perspective 

New viruses in the United States—Hole in the Dike

Surveillance—Pathway AnalysisSurveillance ToolsSolving the ProblemsEnteric Coronaviruses GeneralPrevention/Future VaccinesPDCoV Update

Recent Emergence of Swine Coronaviruses:

A Virologist’s PerspectiveNew viruses in the United States—

Hole in the Dikeo PEDo PDCoVo Porcine TorooNovel PCV2o Entero G

What others are lurking out there??

Solving the Problems

Global Community--Hole in the Dike

Surveillance to be able to conduct Pathway Analysis International National/Hemisphere NAHLN

Surveillance Toolso Scientific Literatureo Traditional-PCRoNext Generation Sequencingo Infectious Virus Vs. qPCR Positive

Bioassay-other

Solving the Problems

Global Community--Hole in the DikeLeadership not Finger Pointing Industry

International Swine Consultants AASV NPB Vaccine Companies

Government USDA DHS

The conference, hosted by the USDA, the U.S. Animal and Plant Health Inspection Service (APHIS), with support from the National Pork Board, the National Pork Producers Council, and the American Association of Swine Veterinarians, will include multiple sessions on nSECDs. Topics will include disease pathways, risk factors for introduction, immunology/vaccinology for nSECDs, as well as the identification of other transboundary swine diseases of concern.

Porcine Epidemic Diarrhea Virus

& Porcine Deltacoronavirus

are Here--Ready or Not

Dick HesseKansas State Veterinary Diagnostic

Laboratory15 September 2014

18

The Disease

Courtesy of Matt Ackerman

Courtesy of Matt Ackerman

20Courtesy of Matt Ackerman

21

Courtesy of Matt Ackerman

Porcine Epidemic Diarrhea Virus (PEDV)

& Porcine Deltacorona Virus (PDCoV):

• Pig-only diseases which do not affect other species or humans and are not a food safety concern.

• PED has been in Europe since 1971, Asian variants are more virulent & traditional vaccines don’t work anymore.

• PDCoV is a new diarrheal disease of swine, we don’t know much about it.

Porcine Epidemic Diarrhea Virus (PEDV)

& Porcine Deltacorona Virus (PDCoV):

• PED incubation time is typically less than 36 hours and virus is shed in feces for 42 days or more.

• PED Age dependent disease severity—Approaching 100% mortality in nursing pigs.

• PDCoV is also age dependent, but mortality in nursing pigs ranges from 20-60%.

• Vomiting and occasional diarrhea in sows and gilts and mild to moderate diarrhea and vomiting in recently weaned pigs.

Porcine Epidemic Diarrhea Virus (PEDV)

& Porcine Deltacorona Virus (PDCoV):

• Laboratory diagnosis is required for definitive confirmation—especially in Older Pigs

• Clinical signs of PED and PDCoV are indistinguishable from the epidemic form of the disease caused by a different porcine coronavirus, Transmissible Gastroenteritis Virus (TGE).

• There is no cross-protection between these coronaviruses.

PED Positive 3 Sept 14

PDCoV Positive 3 Sept 14

What is Next??

Prevention/Future Vaccines

Controlled Infection (Priming??) tissue derived inocula tissue culture derived inocula- bend the

rules??

Vaccines Mucosal/Lactogenic Immunity Priming

Age Gestational Status

Boosting Duration of Immunity

Two USDA Approved Conditional PED Vaccines

In North America:

Zoetis—Classical Killed??

Harrisvaccines--RNA Platform subunit vaccine

Canadian Food Inspection Agency (CFIA) allows import under an emergency permit.

Mucosal Priming Vaccine Production

Courtesy of Matt Ackerman

Mucosal Priming Vaccine Production--BioReactor

32Courtesy of Matt Ackerman

Determination of the sites of tissue localization, routes of viral shedding, duration of virus

carriage, kinetics of antibody response, and potential of aerosol transmission of

Porcine Deltacoronavirus (PDCoV) following inoculation of nursing pigs and their dams.

Koch’s Postulates FulfilledPreliminary Results

PDC0V Team MembersKSVDL--Dick Hesse, Barb Breazeale, Alex Fuller, Joe Anderson, Erin Schirtzinger, Elizabeth Poulsen, Jianfa Bai, Gary Anderson.

UNL--Sarah Vitosh-Sillman, Clayton Kelling, Bruce Brodersen, John Loy, Alan Doster, Chris Topliff.

SDSU—Eric Nelson.

Thanks to the National Pork Board for providing Funding

Housed in five BSL2 isolation rooms at the Life Science Annex at the University of Nebraska.

10 PEDV/PDCoV naïve Bred Sows obtained from a high health commercial source.

Group Treatment # of Animals

A PDCoV oronasal inoculated ~48 (6 litters)

B None—Contact Control 12 (2 from 6 litters)

C None—aerosol transmission controls ~20 (2 litters)

D Negative Controls ~20 (2 litters)

A-Dams PDCoV oronasal inoculated 6

C-Dams None—aerosol transmission controls 2

D-Dams Negative Controls 2

Challenge: Pool of fecal derived virus that was passaged in 2-4 day old pigs.

Kindly provided by Eric Nelson

The inocula had a PDCoV nucleic acid “CT titer” of 22 in a real-time PCR assay.

Nursing pigs and their Dams challenged when pigs were ~2 days of age via intranasal and oral routes with 5 ml of inocula per route.

Day Post Challenge Blood Nasal

SwabFecal Swab

Sequential Sacrifice

Group # Sac

Oral FluidRope

Sample

-2 Sows only Sows only

Sows only    

0 A,B,C,D A,B,C,D A,B,C,D  

1 A-3 + sac A,B,C,D A,B,C,D A-3 D-1  

2 A-3 + sac A,B,C,D A,B,C,D A-3 D-1  

3 A-3 + sac A,B,C,D A,B,C,D A-3 D-1  

4 A-3 + sac A,B,C,D A,B,C,D A-3 D-1  

5 A-3 + sac A,B,C,D A,B,C,D A-3 D-1  

6 A-3 + sac A,B,C,D A,B,C,D A-3 D-1  

7 A,B,C,D A,B,C,D A,B,C,D A-3 D-1  

10 A,B,C A,B,C,D A,B,C,D A-3 D-1  

14 A,B,C,D A,B,C,D A,B,C,D A-3 D-1 A,B,C,D

21 A,B,C,D A,B,C,D A,B,C,D A-3 D-1 A,B,C,D

28 A,B,C,D A,B,C,D A,B,C,D A-3 D-1 A,B,C,D

35 A,B,C,D A,B,C,D A,B,C,D A-3 D-1 A,B,C,D

42 A,B,C,D A,B,C,D A,B,C,D A-12, B-12, C,-12,D-

8 A,B,C,D

Tissues for Histopathology, qPCR and IHC

turbinates, tonsils, esophagus, stomach, duodenum, jejunum (two segments), ileum, cecum, spiral colon, descending colon, trachea, lungs (representative samples from cranial, middle and caudal lobes), liver, spleen, kidney, thymus and submandibular, tracheobronchial, mesenteric and inguinal lymph nodes.

Clinical Observations Following Challenge

• Sows developed soft feces on day 2 post-inoculation and diarrhea on day 3 post-inoculation.

• All sows were clinically normal after day 8 post-inoculation.

• Aerosol contact sows developed soft to diarrheic feces on day 5 of the study and returned to normal after day 8.

• Non-infected control sow remained clinically normal throughout the study.

Clinical Observations Following Challenge

• Inoculated piglets developed soft to profuse diarrheic feces on day 2 post-inoculation, as did the contact controls.

• Morbidity was 100%.

• Mortality was variable among the litters, but was especially high in two of the six inoculated litters.

• All piglets from those two litters were either euthanized due to severe dehydration or had died by day 8 post-inoculation.

Clinical Observations Following Challenge

• Aerosol contact litters developed diarrhea on days 3 and 4.

• All surviving piglets had returned to normal by day 12.

• Non-infected negative control piglets remained clinically normal

Live Animal Fecal Swabs PCRPig ID Nec Day Day 1 Day 2 Day 3 Day 4 Day 5 Day 8 Day 10 Day 14 Day 21 Day 28 Day 35 Day 42

4-8 1.1 40.01-4 1.3 40.05-1 1.3 40.06-8 1.3 38.54-6 2.2 40.0 30.31-1 2.3 40.0 18.94-2 2.3 40.0 15.16-7 2.3 40.0 19.21-2 3.1 40.0 19.1 23.46-3 3.1 40.0 18.94-5 3.2 40.0 17.5 18.26-4 3.2 40.0 21.8 16.51-7 3.3 40.0 16.6 21.13-4 3.3 40.0 18.9 17.54-10 3.3 40.0 17.5 18.41-3 4.3 39.1 19.1 19.2 29.51-8 4.3 40.0 20.9 20.0 17.32-6 4.3 37.7 17.8 16.2 20.83-7 4.3 37.7 15.4 18.1 33.05-3 4.3 40.0 15.4 28.4 32.26-2 5.1 40.0 17.2 19.1 36.76-9 5.3 38.5 16.2 21.5 26.61-5 6.3 40.0 17.8 20.1 32.2 20.13-3 6.3 40.0 17.5 19.7 19.3 22.63-5 7.1 40.0 15.1 16.0 19.1 17.03-6 8.1 40.0 25.6 17.0 21.2 16.51-6 8.3 40.0 17.6 21.7 23.3 31.6 40.06-5 8.3 40.0 30.6 18.1 27.6 16.26-6 8.3 40.0 14.3 36.1 19.5 22.12-3 9.1 40.0 17.0 20.4 23.1 22.7 37.43-8 9.1 40.0 25.0 20.4 26.2 18.1 39.12-7 10.3 40.0 30.3 22.2 30.2 31.3 33.4 35.83-1 10.3 40.0 16.4 19.1 19.4 31.4 33.2 31.74-1 14.3 40.0 19.6 22.7 37.2 24.1 30.1 23.5 35.35-7 14.3 40.0 17.2 18.6 31.8 31.0 36.5 23.7 26.02-9 21.3 40.0 26.9 20.4 33.2 25.1 25.6 26.0 25.1 39.35-4 21.3 40.0 12.6 22.9 22.2 22.2 30.5 22.4 32.4 40.05-5 28.3 40.0 16.0 25.3 18.3 20.7 40.0 34.4 35.8 40.0 40.02-8 35.3 38.7 20.3 17.8 38.3 30.1 32.2 35.5 40.0 40.0 40.0 40.05-2 35.3 40.0 15.5 20.6 40.0 28.9 38.1 35.1 26.0 36.7 40.0 40.02-2 42.3 40.0 28.7 18.1 33.3 38.6 26.7 35.4 40.0 39.3 40.0 40.0 40.02-4 42.3 37.6 21.8 30.6 36.1 34.9 32.8 36.1 40.0 40.0 40.0 40.0 40.02-5 42.3 40.0 14.1 18.3 22.0 31.8 34.9 26.3 25.5 36.8 40.0 40.0 40.03-2 42.3 40.0 16.0 19.9 29.5 18.3 39.1 33.6 40.0 38.0 40.0 40.0 38.54-3 42.3 40.0 18.2 26.1 29.6 39.8 38.2 33.4 31.2 31.0 40.0 40.0 40.04-7 42.3 40.0 17.7 20.3 24.4 36.1 29.3 29.8 24.2 40.0 40.0 40.0 40.05-6 42.3 40.0 15.5 16.0 32.2 33.5 33.7 19.2 40.0 40.0 40.0 40.0 40.05-8 42.3 40.0 14.3 19.1 27.4 23.0 36.5 29.1 37.1 40.0 40.0 40.0 40.0

Dead

/Sacrifi

ced

Tis

su

e P

CR

Pig ID Group Nec Day Blood Serum Trachea Turbinate Tonsil Lung Liver Kidney SpleenLymph Node Thymus

Esophagus Stomach

Des. Colon

Spiral Colon

Small Intestine Cecum

2-1 -1.14-4 -1.14-9 -1.15-11 -1.1 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.06-1 -1.16-10 -1.14-8 A 1.11-4 A 1.3 32.19 34.31 33.5 35.3 40.0 32.4 32.7 36.8 36.4 35.9 37.6 33.6 29.0 34.5 38.8 18.7 23.75-1 A 1.3 31.13 31.63 32.5 36.1 36.7 32.2 29.1 26.6 26.5 23.1 33.4 37.0 32.0 36.2 29.0 16.3 20.56-8 A 1.3 32.15 34.38 36.9 35.6 40.0 31.2 29.8 29.5 37.8 28.9 36.3 36.9 37.4 35.3 29.8 19.6 29.61-9 B 2.2 29.59 28.14 23.7 21.6 23.0 21.5 22.4 17.7 25.1 20.0 23.3 20.1 19.9 18.2 17.6 13.0 19.04-6 A 2.2 27.35 24.3 24.4 31.2 25.1 25.7 22.3 24.8 26.1 25.1 25.8 22.6 27.8 22.9 15.6 17.55-9 B 2.2 33.71 31.52 24.0 20.6 23.3 22.8 26.8 23.3 28.2 19.6 27.7 22.8 21.4 14.9 13.9 13.9 15.71-1 A 2.3 29.17 36.7 25.5 23.1 25.8 22.4 25.4 21.9 21.0 20.5 23.7 24.1 21.9 20.1 17.6 17.1 20.04-2 A 2.3 40 39.2 24.3 23.6 24.0 21.9 24.5 20.9 21.5 20.4 21.9 25.4 20.5 19.0 15.1 16.3 18.46-7 A 2.3 32.05 27.89 23.2 24.5 25.6 25.0 27.8 20.1 31.6 20.7 31.6 24.2 22.2 19.7 16.5 14.3 16.17-10 C 2.3 34.21 32.95 23.8 27.8 25.1 22.2 20.0 18.3 19.5 17.3 22.0 22.7 17.0 14.2 12.4 12.4 15.99-9 D 2.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.01-2 A 3.1 35.1 34.2 35.0 36.9 35.5 30.5 33.8 25.2 40.0 30.5 26.1 22.2 18.9 17.6 20.31-10 B 3.1 33.0 32.6 40.0 31.5 34.3 27.3 33.0 23.3 40.0 33.2 23.4 21.3 19.8 20.2 34.66-3 A 3.1 31.2 32.3 32.6 30.5 32.8 28.6 32.0 26.8 31.0 35.2 24.5 23.0 19.1 18.7 19.84-5 A 3.2 34.67 33.7 30.6 29.1 34.5 31.3 31.5 36.2 28.3 39.5 26.5 30.0 20.9 20.4 17.4 22.06-4 A 3.2 35.51 27.1 26.3 28.8 28.5 33.0 26.9 34.1 22.5 33.6 28.2 24.3 19.7 17.3 15.9 18.67-6 C 3.2 40 35.2 25.4 26.8 24.5 31.2 31.0 36.1 23.3 32.4 26.0 22.6 21.3 18.4 15.8 19.71-7 A 3.3 35.03 37.0 28.9 40.0 34.0 38.9 27.4 36.3 25.6 37.5 37.2 24.5 21.7 18.3 18.5 19.43-4 A 3.3 32.14 31.9 24.7 26.9 31.0 28.4 35.9 30.0 21.6 39.5 26.7 23.7 23.5 22.5 16.8 18.54-10 A 3.3 36.13 33.3 27.8 32.8 32.1 34.2 26.9 34.7 30.4 33.6 29.1 30.6 22.2 20.1 18.6 20.59-5 D 3.3 40 40.0 40.0 40.0 40.0 34.6 40.0 36.2 38.7 40.0 40.0 40.0 40.0 40.0 40.0 40.09RA D 3.3 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.01-3 A 4.3 40 32.55 27.9 32.1 28.2 30.7 27.9 30.2 26.3 29.7 29.3 24.1 19.3 24.1 19.4 16.51-8 A 4.3 40 37.7 34.4 31.4 28.8 34.9 35.2 31.5 33.2 28.2 37.8 27.8 21.5 18.2 20.3 15.0 17.42-6 A 4.3 34.24 36.22 28.6 25.6 35.1 34.3 35.8 27.7 29.5 26.6 32.5 32.5 24.5 20.3 20.2 16.9 19.83-7 A 4.3 33.84 29.99 38.8 28.8 36.1 34.3 31.8 37.8 31.7 28.6 37.1 36.1 25.3 25.1 22.1 19.5 23.35-3 A 4.3 34.69 32.74 35.5 35.1 36.3 36.0 32.2 40.0 34.0 23.5 38.4 33.2 29.7 26.5 24.8 17.6 17.99RB D 4.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 36.7 40.0 40.0 40.06-2 A 5.1 37.6 40.0 35.0 37.2 38.6 33.1 37.5 24.4 37.5 36.0 39.3 30.1 29.3 32.5 27.26-9 A 5.3 40 33.26 38.6 32.2 34.8 34.8 32.8 36.5 34.0 23.2 37.7 34.7 26.7 25.4 21.5 20.1 26.11-5 A 6.3 40 40 40.0 35.9 39.2 40.0 36.4 33.1 35.7 23.5 36.1 37.2 31.9 26.3 20.6 22.2 24.93-3 A 6.3 40 40 34.1 33.1 31.2 35.7 37.4 34.6 35.1 25.2 35.0 30.4 24.3 25.9 17.1 17.08-5 C 6.3 40 40 38.7 35.2 34.2 40.0 32.8 40.0 39.5 35.0 40.0 32.8 28.3 25.6 24.7 17.1 20.49-8 D 6.3 40 40 40.0 40.0 40.0 39.1 40.0 40.0 40.0 40.0 40.0 39.4 40.0 40.0 40.0 36.9 40.03-5 A 7.1 40.0 35.6 34.9 35.4 38.4 34.6 39.2 25.1 39.4 40.0 40.0 33.4 26.5 21.1 36.83-6 A 8.1 40.0 34.2 40.0 34.9 40.0 38.5 40.0 29.9 37.0 40.0 29.5 25.4 26.1 28.0 22.98-2 C 8.1 40.0 38.0 40.0 38.9 39.1 35.2 40.0 32.4 40.0 38.6 33.2 29.9 32.4 23.0 23.08-4 C 8.1 40.0 39.2 40.0 39.5 37.9 40.0 35.3 28.8 40.0 38.6 28.6 30.4 30.8 27.4 26.18-7 C 8.1 36.1 34.0 32.0 34.9 33.0 28.3 40.0 31.3 35.6 40.0 29.0 24.7 21.6 26.3 23.91-6 A 8.3 40 39.03 40.0 35.8 39.3 36.8 40.0 40.0 28.2 40.0 40.0 40.0 40.0 38.7 40.0 40.06-5 A 8.3 40 40 35.4 30.1 34.5 39.2 38.5 38.0 40.0 28.9 40.0 35.2 20.0 30.4 25.2 25.4 20.46-6 A 8.3 40 40 33.3 33.3 31.8 35.4 40.0 34.9 38.0 26.1 35.8 30.9 25.8 33.6 22.1 25.3 26.42-3 A 9.1 40 40.0 39.2 40.0 40.0 40.0 40.0 37.5 25.9 40.0 36.7 35.7 40.0 37.3 35.1 35.73-8 A 9.1 40 37.3 36.9 36.7 35.3 40.0 38.9 38.2 33.1 38.9 37.3 40.0 34.5 33.3 34.8 31.42-7 A 10.3 40 40 40.0 37.5 40.0 40.0 40.0 40.0 40.0 32.7 40.0 40.0 39.1 40.0 36.5 25.3 36.73-1 A 10.3 40 40 38.4 40.0 36.7 37.3 40.0 38.7 40.0 31.8 40.0 36.9 38.8 37.8 38.6 28.3 40.09-2 D 10.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.04-1 A 14.3 40 40 40.0 38.1 36.7 40.0 40.0 40.0 40.0 28.4 40.0 40.0 37.2 35.5 33.5 25.9 35.35-7 A 14.3 40 40 36.8 35.7 35.7 33.3 38.8 32.9 35.8 26.7 37.1 36.0 34.1 29.0 25.8 28.2 26.37-5 C 14.3 40 40 32.0 31.1 31.9 33.2 36.4 32.5 35.0 31.0 36.8 32.2 35.8 24.8 24.1 28.0 23.58-3 C 14.3 40 40 40.0 40.0 38.2 38.7 40.0 40.0 40.0 30.3 40.0 40.0 40.0 40.0 40.0 32.7 40.09-6 D 14.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0

Dead/Sacrificed Tissue PCR

Pig ID Group Nec Day Blood Serum Trachea Turbinate Tonsil Lung Liver Kidney SpleenLymph Node Thymus

Esophagus Stomach

Des. Colon

Spiral Colon

Small Intestine Cecum

4-1 A 14.3 40 40 40.0 38.1 36.7 40.0 40.0 40.0 40.0 28.4 40.0 40.0 37.2 35.5 33.5 25.9 35.35-7 A 14.3 40 40 36.8 35.7 35.7 33.3 38.8 32.9 35.8 26.7 37.1 36.0 34.1 29.0 25.8 28.2 26.37-5 C 14.3 40 40 32.0 31.1 31.9 33.2 36.4 32.5 35.0 31.0 36.8 32.2 35.8 24.8 24.1 28.0 23.58-3 C 14.3 40 40 40.0 40.0 38.2 38.7 40.0 40.0 40.0 30.3 40.0 40.0 40.0 40.0 40.0 32.7 40.09-6 D 14.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.02-9 A 21.3 40 40 40.0 38.5 34.5 40.0 40.0 40.0 36.1 31.4 40.0 36.9 35.3 40.0 40.0 28.4 35.95-4 A 21.3 40 40 40.0 40.0 40.0 40.0 40.0 38.5 40.0 29.9 40.0 40.0 40.0 40.0 37.9 30.2 40.07-2 C 21.3 40 40 40.0 40.0 36.2 40.0 40.0 40.0 37.9 27.1 40.0 40.0 38.7 36.7 36.6 25.0 32.68-1 C 21.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 28.0 40.0 40.0 40.0 40.0 40.0 26.4 40.09-1 D 21.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.05-5 A 28.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.7 40.0 40.0 40.0 40.0 40.0 39.6 38.97-7 C 28.3 40 40 40.0 40.0 39.1 40.0 40.0 40.0 40.0 35.5 40.0 40.0 40.0 40.0 40.0 30.1 35.48-8 C 28.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.7 40.0 40.0 40.0 40.0 40.0 31.0 40.09-3 D 28.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0S2 A 35.3 40 40.0 40.0 40.0 31.6 40.0 38.1 35.4 40.02-8 A 35.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 39.1 30.8 40.0 40.0 40.0 40.0 40.0 32.2 40.02-11 B 35.3 40.0 40.0 38.3 40.0 38.9 40.0 40.0 31.4 40.0 40.0 40.0 40.0 40.0 30.5 40.0S3 A 35.3 40 40.0 40.0 40.0 29.7 40.0 40.0 40.0 40.0 40.0S4 A 35.3 40 38.5 40.0 40.0 32.4 40.0 40.0 40.0 40.0 40.0S5 A 35.3 40 39.0 40.0 37.6 31.3 40.0 40.0 40.0 37.9 40.05-2 A 35.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.6 40.0 40.0 40.0 40.0 40.0 31.2 40.0S6 A 35.3 40 40.0 40.0 40.0 27.3 40.0 38.9 40.0 37.3 40.0S7 C 35.3 40 40.0 40.0 40.0 32.6 40.0 40.0 40.0 31.2 40.07-3 C 35.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.3 40.0 40.0 40.0 40.0 40.0 32.3 40.07-8 C 35.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.0 40.0 40.0 40.0 39.3 29.2 29.8 40.08-10 C 35.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 38.0 40.0 40.0 40.0 40.0 40.0 33.1 39.6S9 D 35.3 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.09-4 D 35.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0S1 A 36.3 40 38.6 40.0 40.0 29.1 40.0 36.1 34.6 40.0 40.0S8 C 36.3 40 40.0 40.0 36.8 34.6 40.0 40.0 40.0 32.7 38.22-2 A 42.3 40 40 40.0 40.0 38.8 40.0 40.0 40.0 40.0 34.8 40.0 40.0 40.0 40.0 40.0 34.2 40.02-4 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 34.2 40.0 40.0 40.0 40.0 40.0 33.8 40.02-5 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.6 40.0 40.0 40.0 40.0 39.4 32.8 40.02-10 B 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 35.3 40.0 40.0 40.0 40.0 40.0 32.4 40.03-2 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 35.3 40.0 40.0 40.0 40.0 40.0 31.2 40.03-9 B 42.3 40 40 40.0 38.5 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 33.8 40.03-10 B 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 33.2 40.0 40.0 40.0 40.0 40.0 40.0 40.04-3 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 34.5 40.0 40.0 38.4 40.0 40.0 33.6 40.04-7 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 33.2 40.0 40.0 40.0 40.0 40.0 31.4 40.05-6 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 34.4 40.0 40.0 40.0 37.0 40.0 32.8 40.05-8 A 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 32.0 40.0 40.0 40.0 40.0 40.0 34.5 38.85-10 B 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 33.8 40.0 40.0 40.0 40.0 35.5 40.07-1 C 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 37.6 40.0 40.0 40.0 40.0 40.0 35.5 40.07-4 C 42.3 40 40 40.0 40.0 40.0 39.0 40.0 40.0 40.0 35.9 40.0 40.0 40.0 40.0 40.0 34.4 40.07-9 C 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 33.8 40.0 38.7 40.0 40.0 40.0 32.8 40.08-6 C 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 33.8 40.0 40.0 40.0 40.0 40.0 40.0 40.08-9 C 42.3 40 40 40.0 40.0 37.5 40.0 40.0 40.0 40.0 32.7 40.0 40.0 40.0 40.0 40.0 33.3 40.09-7 D 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.09-10 D 42.3 40 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0

Oral Fluid PCR

Oral Fluid Day 14 Day 21 Day 28 Day 35 Day 39Litter 1Litter 2 30.1 32.1 30.3 40.0 38.5Litter 3 33.7 34.7 37.0 38.7 40.0Litter 4 34.5 35.2 40.0 40.0 40.0Litter 5 31.3 31.8 40.0 40.0 40.0Litter 6Litter 7 28.1 36.9 36.6 40.0 40.0Litter 8 35.1 35.4 34.5 40.0 37.6Litter 9 40.0 40.0 40.0 40.0 40.0

Sow Data Following PDCoV Challenge

Viremia

Fecal Shedding

Nasal Shedding

Day 35Tissue PCR

Serum PCRPig ID Day 8 Day 14 Day 21 Day 28 Day 35

S1 40.0 40.0 40.0 40.0 40.0S2 40.0 40.0 40.0 40.0 40.0S3 40.0 40.0 40.0 40.0 40.0S4 40.0 40.0 40.0 40.0 40.0S5 40.0 40.0 40.0 40.0 40.0S6 40.0 40.0 40.0 40.0 40.0S7 40.0 40.0 40.0 40.0 40.0S8 40.0 40.0 40.0 40.0 40.0S9 40.0 40.0 40.0 40.0 40.0

Fecal PCRPig ID Day 1 Day 2 Day 3 Day 4 Day 5 Day 8 Day 10 Day 14 Day 21 Day 28 Day 35

S1 40.0 30.1 24.6 22.8 23.2 24.0 27.3 32.8 37.2 40.0 32.5S2 40.0 30.5 23.9 24.0 22.5 23.4 18.9 28.4 36.6 40.0 40.0S3 40.0 29.9 30.0 25.0 27.4 19.8 21.2 33.0 38.4 40.0 40.0S4 40.0 40.0 31.5 33.8 40.0 27.2 24.5 35.6 40.0 40.0 40.0S5 40.0 29.3 21.8 26.5 20.2 27.9 26.9 36.1 40.0 40.0 40.0S6 40.0 32.0 32.8 23.3 23.9 22.8 25.4 27.2 34.6 40.0 40.0S7 40.0 25.6 28.4 24.1 27.3 29.9 27.9 35.6 36.8 40.0 38.8S8 40.0 37.0 29.1 20.2 19.4 21.4 24.4 31.7 35.4 33.4 40.0S9 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0

Nasal PCRPig ID Day 1 Day 2 Day 3 Day 4 Day 5 Day 8 Day 10 Day 14 Day 21 Day 28 Day 35

S1 40.0 28.0 29.2 29.1 35.9 37.6 40.0 38.9 40.0 40.0 40.0S2 40.0 27.4 29.3 32.2 35.4 35.1 34.7 37.0 40.0 40.0 40.0S3 40.0 31.1 29.3 33.1 33.3 37.4 35.2 40.0 38.6 40.0 40.0S4 40.0 33.5 33.2 36.8 39.7 40.0 36.4 40.0 40.0 40.0 40.0S5 40.0 32.1 40.0 40.0S6 40.0 32.6 30.2 32.1 34.5 29.6 36.3 40.0 40.0 40.0 40.0S7 40.0 29.8 31.7 34.3 30.8 40.0 34.0 35.5 40.0 40.0 40.0S8 40.0 36.1 30.1 32.2 33.7 36.9 39.3 40.0 40.0 40.0 39.3S9 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0

Pig ID Serum Tonsil Liver SpleenLymph Node Stomach

Des. Colon

Spiral Colon

Small Intestine Cecum

S1 40 38.6 40.0 40.0 29.1 40.0 36.1 34.6 40.0 40.0S2 40 40.0 40.0 40.0 31.6 40.0 38.1 35.4 40.0S3 40 40.0 40.0 40.0 29.7 40.0 40.0 40.0 40.0 40.0S4 40 38.5 40.0 40.0 32.4 40.0 40.0 40.0 40.0 40.0S5 40 39.0 40.0 37.6 31.3 40.0 40.0 40.0 37.9 40.0S6 40 40.0 40.0 40.0 27.3 40.0 38.9 40.0 37.3 40.0S7 40 40.0 40.0 40.0 32.6 40.0 40.0 40.0 31.2 40.0S8 40 40.0 40.0 36.8 34.6 40.0 40.0 40.0 32.7 38.2S9 40 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0 40.0

HistopathologyVirus Localization

• Histological lesions of the GI tract were extensive, mainly confined to the small intestine.

• PDCoV was demonstrated via IHC in the villi of the small intestine.

• Detection of virus localization in other tissues is currently underway.

Histopathology/Virus Localization

Day 1 post inoculation

ImmunohistochemistryVirus Localization

Courtesy of Bruce Brodersen and Sarah Vitosh-Sillman.

Detection Antibody provided by Eric Nelson

IFA and SN Response followingInoculation

• All sows seroconverted to PDCoV (IFA & SN) around D-14 post inoculation except the Negative Control group.

• All surviving piglets seroconverted to PDCoV (IFA & SN) around D-14 post inoculation except the Negative Control group.

Thanks To:• NVSL—Sabrina Swenson &

Melinda Jenkins-Moore for providing cell culture adapted PDCoV virus, culturing methods and sharing assay results.

Questions

Questions