INFLUENCE OF TYPE 2 BOVINE VIRAL DIARRHEA VIRUS NPRO ON ENHANCEMENT OF BOVINE RESPIRATORY SYNCYTIAL VIRUS REPLICATION MEDIATED BY ANTAGONISM OF HOST CELL INTERFERON TYPE I RESPONSES

Bovine viral diarrhea virus (BVDV) is a member of the genus Pestivirus, Family Flaviviridae. The virus can infect many species of animals of the order Artiodactyla. The BVDV genome encodes an auto protease, Npro, that degrades interferon regulatory factor-3 (IRF-3) reducing type I interferon (IFN-I) production from host cells. Bovine respiratory syncytial virus (BRSV) is a member of the genus Pneumovirus, Family Paramyxoviridae. Concurrent infection with BVDV and BRSV causes more severe respiratory and enteric disease than infection with either virus alone. Our hypothesis was that Npro modulates the innate immune responses to BVDV infection and enhances replication of BVDV or BRSV co-infection. The noncytopathic BVDV2 viruses NY93/c N- Npro 18 EGFP (a mutant with modified Npro fused with enhanced green fluorescent protein), NY93 infectious clone (NY93/c), wild-type NY93-BVDV2 (NY93-wt), and BRSV were evaluated in this study. The objectives of this study were: (1) to characterize the replication kinetics and IFN-I induction in Madin-Darby bovine kidney (MDBK) cells following infection with each of the BVDV isolates, and (2) to characterize the influence of BVDV-mediated IFN-I antagonism on enhancement of BRSV replication in bovine turbinate (BT) cells. NY93/c N- Npro 18 EGFP replicated 0.4 – 1.6 TCID50 logs lower than NY93-wt in MDBK cells. NY93/c N- Npro 18 EGFP-infected MDBK cells synthesized IFN-I significantly higher than NY93/c- and NY93-wt-infected MDBK cells. BT cells co-infected with NY93/c N- Npro 18 EGFP/BRSV or NY93-wt/BRSV were evaluated to determine the effects of co-infection on BRSV replication and IFN-I induction in BT cells. BRSV RNA levels in NY93-wt/BRSV co-infected BT cells were 2.49, 2.79, and 2.89 copy number logs significantly greater than in NY93/c N- Npro 18 EGFP/BRSV co-infected BT cells on days 5, 7, and 9 post-infection, respectively. BVDV RNA levels in NY93/c N- Npro 18 EGFP-infected BT cells were 1.64 – 4.38 copy number logs lower than in NY93-wt-infected BT cells. NY93/c N- Npro 18 EGFP single and co-infected BT cells synthesized IFN-I significantly higher than NY93-wt single and co-infected BT cells. In summary, these findings suggest: (1) NY93/c N- Npro 18 EGFP BVDV2 induced higher levels of IFN-I than BVDV2-wt and may be useful as a safer, replicating BVDV vaccine, and (2) Enhancement of BRSV infection by BVDV co-infection is mediated by antagonism of IFN-I.

SCWDS Briefs: Volume 25, Number 2 (July 2009)

Table of Contents: Piroplasmosis Hits Missouri Horses Cytauxzoon felis in Wild Felids SCIF & Mossy Oak Fund HD Research Hardware Disease in a Key Deer Wildlife Poisoning in Kansas Swine Brucellosis Infects Hog Hunters SCWDS Personnel Changes NWHC Has New Director New USDI Publication on Bats Recent SCWDS Publications Available

SCWDS Briefs: Volume 25, Number 3 (October 2009)

Table of Contents: Clams & AI Virus SCWDS Funded to Study WNS USDA Program Changes Proposed TB in Captive Cervids CWD Prions in Deer Feces TWS Position Statement on Lead WDA Awards For SCWDS Staff New Brochure for Hog Hunters

SCWDS Briefs: Volume 24, Number 3 (October 2008)

Table of Contents: Piroplasmosis in Florida Horses Bovine TB Update Newcastle Disease in Cormorants Frog Virus in a Box Turtle SCWDS Vesicular Stomatitis Research Update Faculty and Staff Changes at SCWDS Lead Study Results Reported Lead Ammo and Tackle Review Our Energizer Bunny

SCWDS Briefs: Volume 23, Number 4 (January 2008)

Table of Contents: Orbiviruses New & Old - What Do We Need to Know? Orbivirus Vector Surveys Studies on H5N1 HPAI Virus in Swans and Geese Dr. Justin Brown Awards Impacted Turkey Gizzard Regional Disease Workshops in 2008-09 Dove Disease Research at SCWDS Some Staff Changes