Maintenance of Bovine viral diarrhea virus in whitetailed

Maintenance of Bovine viral diarrhea virus in white-tailed deer (Odocoileus virginianus) following exposure to persistently infected cattle T. Passler 1, P. H. Walz 1, S. S. Ditchkoff 2, M. D. Givens 1, R. W. De. Young 3, K. V. Brock 1 1 Departments of Clinical Sciences and Pathobiology, Auburn University, Auburn, Alabama, USA 2 School of Forestry and Wildlife Sciences, Auburn University, Auburn, Alabama, USA 3 Caesar Kleberg Wildlife Research Institute, Texas A&M University-Kingsville, Texas, USA

Experimental Design Cohabitation of BVDV-negative calves with deer Cohabitation of PI cattle with deer Mar 8 Capture of Deer (Dec 1 -18, 2006) May 7 PI deer (17 A) housed at CDRF Jan 1, 2008 Birth and Testing of Fawns (Jul-Aug, 2007) Mating of deer in captivity (Jan 1 -21, 2007) Capture of pregnant deer (Feb 25 -Apr 10) Birth and Testing of Fawns (Jul-Aug, 2008) 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

October 1, 2008: BVDV infections of white-tailed deer (Odocoileus virginianus)

BVDV AU 526 nasal swab titer Entry: 3. 5 x 105 CCID 50/ml Exit: 6. 2 x 104 CCID 50/ml BVDV KY 16 nasal swab titer Entry: 3. 5 x 105 CCID 50/ml Exit: 1. 1 x 105 CCID 50/ml 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

• 7 female white-tailed deer exposed to 2 PI cattle for 60 days • All 7 delivered fawns based upon maternity testing • Twin stillborn fetuses • Fawns were isolated and hand-raised 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

Pertinent Data on Fawns Born after Dams Were Exposed to 2 Cattle PI with BVDV Fawn ID Virus isolation RT-n. PCR ELISA Serum WBC Nasal swab Tissues Serum Tissues Ear notch 17 A ― + ― ND + ND ND 18 A† ― ND ― + ND 18 B† ― ND ― + ND 19 A ― ― ― ND 19 B ― ― ― ND ― 21 A ― ― ― ND 21 B ― ― ― 23 A ― + 24 A ― 24 B 25 A IHC Ear notch Virus neutralization Tissues AU 526 KY 16 ND ND ND + + ND ND ― ― ND 16 16 ND ― ― ND 128 ― ND ― ― ND 32, 768 65, 536 ND ― ― ND 4, 096 131, 072 + ― + + + ND ND ― ― ND 64 256 ― ― ― ND 128 256 ― ― ― + + ND ND + + 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

4 of 7 pregnancies resulted in PI white -tailed deer Acute Infection Fetal Infection Persistent Infection

Exposure of pregnant deer to PI deer • Fawn 17 A: PI with BVDV 1 b AU 526 • Six pregnant does – Wanted pregnancies between 30 -70 days – February 25 – April 10 • Cohabitation for most of gestation – Fawn 17 A found dead on July 10 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

Data on Fawns Born after Dams Were Exposed to Fawn 17 A (PI) Fawn ID RT-PCR Serum VI WBC VI Nasal swab IHC Ear notch VN titer 13 ― ― 2048 14 ― ― 2048 15 ― ― 1024 16 ― ― 2048 17 ― ― 1024 18 ― ― 4096 19 + ― ― ― 1024 20 ― ― 2048 21 + + ― + 8192 22 ― ― 2048 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

August 1, 2008: RT-n. PCR Positive August 28, 2008: RT-n. PCR Positive January 9, 2009: RT-n. PCR Negative Skin IHC: Negative 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

Conclusions 1. Transmission of BVDV from PI cattle to whitetailed deer – Transplacental infection in 5/7 does 2. Transmission of BVDV from PI deer to incontact deer – High antibody titers in fawns • Transplacental exposure versus passive transfer? – One PI white-tailed deer (Fawn 21) • Level of shedding of Fawn 17 A or time of exposure? 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

GN 17 A 26 A 23 A 25 A 18 A 25 -Aug-06 28 -Aug-07 15 -Aug-07 21 -Aug-07 04 -Aug-07 31 -Jul-07 Calculated Conception (205 day gestation) 01 -Feb-06 04 -Feb-07 22 -Jan-07 28 -Jan-07 11 -Jan-07 01 -Feb-07 (195) 18 B 31 A (21) 32 B (19) 32 A (20) 19 A 19 B 21 A 21 B 24 A 24 B 35 A 35 B 36 A 36 B 33 A 33 B 34 A 31 -Jul-07 13 -Aug-08 01 -Aug-08 30 -Jul-07 26 -Aug-07 06 -Jul-08 01 -Aug-08 16 -Aug-08 01 -Feb-07 (195) 21 -Jan-08 09 -Jan-08 06 -Jan-07 02 -Feb-07 12 -Dec-08 09 -Jan-08 24 -Jan-08 Fawn ID DOB Intranasal PI Cow Calculated Fetal Age at Initial Exposure 52 32 51 39 56 51 PI Cow PI Deer PI Cow PI Cow PI Deer PI Deer 51 46 71 71 61 61 34 34 109 119 75 75 68 Method of Exposure Singlet or Twin BVDV Strain Singlet Singlet Twin (SB) BVDV 2 PA 131 BVDV 1 AU 526 BVDV 1 KY 16 Twin (SB) Singlet Twin Twin Twin Twin Singlet BVDV 1 KY 16 BVDV 1 AU 526

DOB Calculated Conception (205 day gestation) Method of Exposure GN 25 -Aug-06 01 -Feb-06 Intranasal 17 A 28 -Aug-07 04 -Feb-07 PI Cow 26 A 15 -Aug-07 22 -Jan-07 Intranasal 23 A 21 -Aug-07 28 -Jan-07 PI Cow 25 A 04 -Aug-07 11 -Jan-07 PI Cow 18 A 31 -Jul-07 01 -Feb-07 (195) PI Cow 18 B 31 -Jul-07 01 -Feb-07 (195) PI Cow 31 A (21) 13 -Aug-08 21 -Jan-08 PI Deer Fawn ID Calculated Fetal Age at Initial Exposure 52 32 51 39 56 51 51 46 Singlet or Twin BVDV Strain Singlet BVDV 2 PA 131 Singlet BVDV 1 AU 526 Twin (SB) BVDV 1 KY 16 Singlet BVDV 1 AU 526

Fawn ID DOB Calculated Conception (205 day gestation) 32 B (19) 01 -Aug-08 09 -Jan-08 PI Deer 32 A (20) 01 -Aug-08 09 -Jan-08 PI Deer 19 A 30 -Jul-07 06 -Jan-07 PI Cow 19 B 30 -Jul-07 06 -Jan-07 PI Cow 21 A 30 -Jul-07 06 -Jan-07 PI Cow 21 B 30 -Jul-07 06 -Jan-07 PI Cow 24 A 26 -Aug-07 02 -Feb-07 PI Cow 24 B 26 -Aug-07 02 -Feb-07 PI Cow 35 A 06 -Jul-08 12 -Dec-08 PI Deer 35 B 06 -Jul-08 12 -Dec-08 PI Deer 36 A 06 -Jul-08 12 -Dec-08 PI Deer 36 B 06 -Jul-08 12 -Dec-08 PI Deer 33 A 01 -Aug-08 09 -Jan-08 PI Deer 33 B 01 -Aug-08 09 -Jan-08 PI Deer 34 A 16 -Aug-08 24 -Jan-08 PI Deer Method of Exposure Calculated Fetal Age at Initial Exposure 71 71 61 61 34 34 109 119 75 75 68 Singlet or Twin BVDV Strain Twin BVDV 1 AU 526 Twin Twin Twin Twin Singlet

Reservoir White-tailed deer as reservoir of BVDV - Criteria to be met: √ - Susceptibility - Disease agent must √ be maintained √ - Disease agent must be shed ? - Sufficient contact between species 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

Acknowledgments Funding provided by: Animal Health Research, Auburn University and an AABP Research Assistantship • • • • Thomas Passler Steve Ditchkoff Dan Givens Kenny Brock Herris Maxwell Terri Hathcock Yijing Zhang Heather Walz Ann Cochran Andrew Eason Leland Neuhring Emily Hanley Randy De. Young George Fincher Wildlife and Veterinary Students 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

Questions? Comments! passlth@vetmed. auburn. edu walzpau@vetmed. auburn. edu 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)

PI white-tailed deer Day of sample collection Serum VI Nasal Swab VI 8/25/06 6. 2 X 105 CCID 50/ml 2 X 106 CCID 50 /ml 9/25/06 6. 2 X 105 CCID 50/ml 2 X 106 CCID 50 /ml 10/23/06 6. 2 x 105 CCID 50/ml 6. 2 x 105 CCID 50 /ml 4 th BVDV Symposium: BVDV infections of white-tailed deer (Odocoileus virginianus)