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.

ENHANCEMENT OF THE FAA’s ON-LINE WILDLIFE AIRCRAFT STRIKE DATABASE WITH AN INTERACTIVE GRAPHICS CAPABILITY

Embry-Riddle Aeronautical University (Prescott, AZ, USA) was awarded a grant from the William J. Hughes FAA Technical Center in October 1999 to develop and maintain a web site dealing with a wide variety of airport safety wildlife concerns. Initially, the web site enabled users to access related topics such as wildlife management (at/near airports), bird identification information, FAA wildlife management guidelines, education, pictures, current news, upcoming meetings and training, available jobs and discussion/forum sections. In April 2001, the web site was augmented with an on-line wildlife strike report (FAA Form 5200-7). Upon submittal on-line, “quick look” email notifications are sent to concerned government personnel. The distribution of these emails varies as to whether there was damage, human injuries/fatalities, and whether feather remains were collected and will be sent to the Smithsonian Institution for identification. In July 2002, a real-time on-line query system was incorporated to allow federal and local government agencies, airport and operator personnel, and USDA and airport wildlife biologists to access this database (which as of June 2005 contains 68,288 researched strike reports added to at a rate of approximately 500 strike reports/month) to formulate strategies to reduce the hazards wildlife present to aviation. To date (June 2005), over 15,000 on-line real-time queries were processed. In June 2004, ERAU was authorized to develop a graphical interface to this on-line query system. Current capabilities include mapping strikes (by species) on the US map, each of the contiguous 48 state maps (with AK and HI being added), and airport diagrams of the major metropolitan airports as well as the next 46 airports with the most reported strikes The latter capability depicts strikes by runway in plan as well as in elevation view. Currently under development is the ability to view time-sequenced strikes on the US map. This extensive graphical interface will give analysts the ability to view strike patterns with a wide variety of variables including species, seasons, migration patterns, etc. on US and state maps and airport diagrams.