Brief Summary of Minutes of Annual Meeting

The meeting was opened with welcomes from Dr. Harry Dickerson, Associate Dean for Research and Graduate Affairs, College of Veterinary Medicine and Director, Veterinary Medical Experiment Station and Dr. Jerry Cherry, Associate Dean for Research, College of Agricultural and Environmental Sciences, and Director, Athens Agricultural Experiment Station. A brief history of college of Veterinary Medicine and Agricultural Experiment Station followed. Business meeting: Updates from the Administrative Advisor: Dr. Asem was unable to attend this meeting He requested that the title and objectives for the new project covering 2006-2011 be submitted to the website following this meeting. Future NC-107 meeting: The group suggested that we may combine the future NC107 meeting with one of the following: (1). The Academy of Veterinary Consultants (beef and feedlot vets). (2). The AABP (mostly dairy, but also some beef vets). (3). The AAVLD (diagnostic Vets) or possibly but not as enthusiastically (4). CRWAD. Because there were only eight participants, it was decided that Dr. Chowdhury (Chair for next year) would send e-mail to every body and get the majority opinion. Because the attendance was low, there was a lengthy discussion about whether or not we should have a continued NC107 meeting. In the end, the consensus was that its a good group and we should not give up. On September 14th (2nd day) the group worked on drafting/revising the objectives for the future NC-107 five year project. New Station representative for Louisiana Dr. Gus Kousoulas was introduced. Officers elected for 2006 were Chair, Dr. Shafiqul Chowdhury, Secretary, Dr. Robert Briggs. Next meeting was left for the outcome of member input. Business Meeting closed on September 14, 2005 at 10:30 AM. New business: The following were developed as the basis of the next project. Title: An integrated approach to the control of bovine respiratory diseases The objectives of the new project are: (1). To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents; (2). To investigate the basic biology, molecular pathogenesis, and immunopathogenesis of polymicrobial infections including important viral and bacterial agents; and (3). To develop management and prevention strategies that incorporate new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss. Station Reports were by the following: Kansas Dr. Shafiqul Chowdhury Louisiana Dr. Gus Kousoulas Oklahoma Dr. Bob Fulton South Dakota Dr. Russ Daly (for Dr. Chris Chase) Wisconsin Dr. Chuck Czuprynski USDA-NADC Dr. Bob Briggs Georgia Dr. Amelia Woolums

OBJECTIVE 1: Identify emerging and re-emerging agents and develop diagnostic methods for bovine respiratory disease (BRD). Oklahoma: Since 2003 we have tested samples from five feedyards in Oklahoma, Colorado, and Kansas to identify bovine viral diarrhea virus (BVDV) persistently infected (PI) cattle. There were 3463 samples tested with 116 positive (3.3%). The range of immunohistochemistry (IHC) positives in the five feedyards was 2.6% to 5.4%. In a recent shipment from one feedyard, 5.1% (8/157) were positive. This is about twice the rate for the past year. Vaccination histories and pen data for morbidity, mortality, treatment costs, number of treatments, and cost of gain are being collected for analysis. Such information will help communicate the adverse effects of PI cattle to feedlot management and ranchers supplying the cattle. BVDV subtypes in diagnostic lab accessions from clinical and necropsy cases, and the distribution of BVDV1a, 1b, and 2A subtypes, was evaluated. The prevalence of BVDV subgenotypes and biotypes was determined from 131 BVDV samples from a diagnostic laboratory. The BVDV samples were segregated into three subgenotypes by sequencing the 5-untranslated region (5UTR). There were 131 BVDV samples represented by 117 noncytopathic (NCP), 11 cytopathic (CP) and 3 cases with both biotypes. The NCP isolates were more common (P<0.05) than CP or NCP/CP. There were more BVDV1b subgenotypes 60/131 (45.8%) than BVDV1a, 37/131 (28.2%) or BVDV2a 34/131 (26.0%). All three subgenotypes were found in persistently infected (PI) cattle and respiratory diseases, both major requests for BVDV diagnosis. Only one of the 131 viruses was genetically similar to the strains in U.S. vaccines. Minnesota: The infectious agents associated with BRD were identified (7/04-6/05). 1. Bacteriology: Bacterial agents identified from bovine pneumonic lungs, nasal swabs, tracheal swabs, transtracheal fluids, and pleural swabs are as follows: Bacteria Isolations H. somnus 61 M. haemolytica 69 P. multocida 97 Mycoplasma sp 72 2. Virology: Viral agents identified by different techniques are as follows: Virus FA IH PCR SN titers <8 Serum HI titers <40 Total IBR - 10 - 419 - 429 PI3 - - - - 58 58 BRSV - 17 - 110 - 127 BVD - 2(tissues) 45(serum) - - 147 11(ear-89 (tissue skin) homogenate) BVD1 - - - 574 - 574 BVD2 - - - 288 - 288 BHV4 1 - - - - 1 Wisconsin: Wisconsin Veterinary Diagnostic Laboratory provided the following data for the year 2004: Summary of WVDL Virology Results (1-01-04 to 12-31-04 ) 2004 #Tested #Positives % BVD PI serum 15,980 22 0.13% BVD-PI Buffy coat 1142 30 2.6 % BVD ear notch ELISA 24,422 144 0.58% BVD-BC acute 269 13 4.8% BVD serum ELISA 1633 11 0.67% Lung tissues VI-BVD 188 12 6.4% Lung tissues VI-IBR 188 1 0.53% FA Frozen section BVD lung 1215 37 3.0% FA Frozen section IBR lung 592 47 7.9% BRSV IHC positives 20 1 ** BRSV SN 474 361 ** PI3 3 0 0% Iowa: Work continues on the development and validation of ELISA serology methods to assess Mycoplasma bovis exposure. An ELISA test based on measuring sero-responses to an immunosuppressive peptide of M. bovis may allow differentiation of vaccinal responses from responses to infection. South Dakota: A. Diagnostic findings: The infectious agents associated with bovine respiratory disease complex were monitored (7/04-6/05): 1. Bacteriology: isolates from bovine pneumonic lungs, tracheal swabs, and nasal swabs are as follows: Organism Isolations M. haemolytica 80 P. multicida 104 H. somnus 63 2. Virology: Viral agents from bovine pneumonic lungs (455 isolation attempts): Virus Virus Isolations Fluorescent Antibody BVDV-NCP 23 BVDV-CP 13 BHV-1 4 BRSV 0 19 BHV-4 (DN-599) 8 3. BVDV Genotyping: 79% of field isolates were BVDV type 2, 21% were type 1. 4. Immunoperoxidase BVDV outgrowth test for the detection of BVDV PI identified 5 positives of 7470. 5. Immunohistochemistry: IHC ear notch BVDV tests were conducted on 23,477 samples with 306 positives (1.3%). In 2004 there were 21,048 samples, 150 positives (0.7%). ELISA ear notch BVDV tests were conducted on 4,419 samples with 9 positives (0.2%). 6. Molecular Diagnostics: PCR tests were done for BVDV and 465 tests (3050 samples) were tested and 53 positives were detected. Typing was done on 91 PCR and VI isolates and 23% type 1 and 77% type 2. Georgia: The prevalence of Mycoplasma bovis in nasal swabs collected from cattle at arrival to backgrounding/stocker operations in Georgia was evaluated. In 9 operations, 87% of cattle were positive for mollicutes by PCR, while only 2% were positive for M. bovis. Fifteen percent were positive for any mycoplasma-type organism by culture, while 2% were positive for M. bovis by culture with PCR for specific identification. Thus herd level prevalence of M. bovis shedding was high but the animal-level prevalence was low, with between 2% - 6% of animals shedding on affected operations. Michigan: A rapid detection of BVDV in cell culture media using a biosensor has been demonstrated. Detection of cattle persistently infected with BVDV using ear notches was most consistent when compared to serum and nasal swabs. Further development may improve this tool for the rapid and economical identification of BVDV PI cattle. Bronchopneumonia continues to be the major cause of feedlot morbidity. The major pathogen associated with bronchopneumonia morbidity in Michigan appears to be Mycoplasma bovis. BVDV is often found in conjunction with M. bovis as well as other respiratory pathogens. NADC: Characterization of goat adenovirus isolates from Minnesota and Canada indicate that there may be across-species antigenic relationships among the ruminant Mastadenovirus, although they differ at the genetic level. Consensus primers for dependovirus and autonomous parvoviruses were developed to help characterize goat adenovirus/dependovirus isolates recovered from a recent outbreak to explore the their role in pathogenicity. Several widespread outbreaks of severe acute BVD have been traced to a single strain of BVDV. Unlike most strains which are low virulence this strain, which is high virulence, spread quickly from herd to herd and caused high mortality. This information is important to producers because the explosive spread of this virus suggests that these outbreaks must be controlled differently than typical outbreaks. We developed a research model that uses cultured cells rather that live animals to differentiate between high and low virulence viruses. This model will reduce the need to use live animals and will significantly cut the cost and difficulty of studying virulence in BVDV strains. OBJECTIVE 2: Characterize mechanisms and intervention targets in pathogenesis of BRD at the molecular, cellular, and host level. Minnesota: The site within the bovine CD18 required for LktA binding and its biological effects was mapped. We created bovine (B) x human (H) chimeric CD18 constructs by domain swapping the extracellular portion of human CD18 with corresponding sequences from the bovine CD18. The constructs were co-expressed with bovine CD11a in K562 cells, and resistance to LktA monitored. The results demonstrate that the binding site for LktA is contained within 500 to 600 amino acid residues of the extracellular region of bovine CD18. Mississippi: Integrated functional genomics was used to determine the effect of sub-lethal antibiotics on bacteria. Prophylactic use of antibiotics significantly reduces morbidity and mortality due to BRD. To determine the underlying mechanism of action of sub-minimum inhibitory concentrations (sub-MIC) of chlortetracycline (CTC) and CTC-sulfamethazine combination (CTC+SMZ), we compared the proteome of respiratory pathogen Mannheimia haemolytica grown in the presence of ¼ MIC of both antibiotic preparations to that obtained without the antibiotics. To analyze protein expression we used two dimensional liquid chromatography and in-line electospray ionization tandem mass spectrometry (2-D LC ESI MS2). When M. haemolytica was cultivated in the presence of ¼ MIC of CTC and CTC+SMZ, expression of 85 proteins was significantly altered by at least one treatment. The most notable sub-MIC effect was a significant decrease in the expression of leukotoxin A. Reduction in leukotoxin expression could be one of the molecular mechanisms responsible for the efficacy of these antibiotics. This finding provides a plausible explanation for the efficacy of prophylactic use of CTC in cattle. We showed that differential detergent fractionation (DDF) analysis of bovine monocytes reveals proteins related to antigen pattern recognition, uptake and presentation to immunocompetent lymphocytes. Recently, we have identified MR as the important receptor involved in endocytosis in bovine monocytes. We have also demonstrated that antigen uptake through MR-mediated endocytosis, an important APC function, is affected in monocytes in the early stage of BVDV infection during the first 24 hrs. In our research, we identified the following proteins related to antigen uptake: the galactose binding lectin, MyD-1, clathrin light chain A and MR precursor. We also identified 8 proteins related to cytoskeleton such as actin, beta actin and gamma actin. Wisconsin: How VP22 of BHV-1 enters the nucleus is unknown. No classic nuclear localization signal (NLS) motif has been identified. To identify the signal sequence directing nuclear accumulation, a series of truncations, internal deletions and point mutations were constructed. Evaluation of cells transfected with VP22 constructs indicated that a sequence of 103 residues is necessary and sufficient for nuclear localization. This sequence is conformation sensitive, in contrast to the classical sequential NLS. VP22 co-immunoprecipitated with histone H4, suggesting that interaction with histone H4 may contribute to the nuclear retention. We investigated intracellular pathways that result in bovine leukocyte death following exposure to M. haemolytica LKT. We find that LKT activates caspase-9 more than caspase-8, suggesting involvement of the mitochondrial pathway of apoptosis in LKT-mediated cell death. The anti-apoptotic protein Bcl-2 decreased, while the pro-apoptotic proteins Bax and Bad increased, following in vitro incubation of cells with LKT . Phosphorylation of Akt-1, a signaling protein downstream of LFA-1 that usually leads to cell survival, was decreased following LKT exposure. In addition we found evidence that the LKT is internalized, associates with actin stress fibers, makes it way to the mitochondria, and binds to the outer mitochondrial membrane. Mechanisms by which Haemophilus somnus causes the vasculitis that characterizes H. somnus infections were studied. We published evidence that H. somnus, but not its LOS, can cause platelet aggregation in vitro. H. somnus and its LOS, as well as H. somnus activated platelets, are capable of up-regulating tissue factor and ICAM-1 (CD54) and downregulating thrombomodulin on bovine pulmonary artery and brain microvascular endothelial cells, as determined by flow cytometry and Western blot. These changes may contribute to the vasculitis and thrombosis seen in H. somnus infections. The most severe manifestation of H. somnus infection is thrombomeningoencephalitis (TEM). Cerebral microvascular endothelial cells are the primary cellular constituent of the blood brain barrier, which protects the central nervous system from inflammation and infection. Using primary cell culture and Haemophilus somnus, we have developed an in vitro model to study the sequence of events and mediators involved in the dysfunction of the blood brain barrier in bacterial infection. We have investigated the ability of H. somnus to attach to and invade primary cerebral microvascular endothelial cells (CMVECs). Attachment occurs in a dose dependent manner, but invasion is rare. Binding of H. somnus to CMVECs appears to be inhibited by heparin, suggesting a role for glycosaminoglycans. CMVECs up-regulate TNF-£\, tissue factor, and ICAM-1 expression, and down-regulate the expression of occludin, a tight junction protein, following exposure to H. somnus. CMVECs exposed to H. somnus also undergo apoptosis. These alterations could play a role in the development of vasculitis, thrombosis and inflammation that results in blood brain barrier dysfunction and bacterial invasion of the brain South Dakota: Two new molecular tools, toll like receptor (TLR) 2 and 4 real time PCR assays, were developed for molecular pathogenesis studies. Monocyte-derived macrophages (MDM) were infected with BVDV strains of different virulence. mRNA for TLR-2 and -4 of virus-infected MDMs was quantified by RT-PCR using molecular beacons. After 12 hrs of infection, a BVDV strain from persistent infection significantly (p<0.05) decreased the amount of TLR-2 mRNA as compared to the mock and other strains. However, this virus has only minimal effect on TLR-4 mRNA Thus, BVDV may modify the innate immune recognition by influencing the expression of toll-like receptors. Thirty-nine calves from a single farm were transported to a university facility in September 2004. Subsequent testing revealed that 36 of the animals were PI with BVDV type 2a. These animals were analyzed over a 10-month period for virological and serological changes. NCP-BVDV was isolated from all the calves and the 5 UTR and E2 region were analyzed. The results indicate that the 5UTR was similar among all calves but one group of 7 calves had conserved changes in the E2 region. Twenty-four of the calves succumbed to mucosal disease during the experiment and CP-BVDV virus was isolated from 18 calves. The CP-BVDV is currently being sequenced to determine mutation differences. Six of the PI animals developed SN titers against both type 1 and type 2 calves. There was no relationship between the development of antibody and onset of mucosal disease. These results indicate that within a population of PI infected animals that the amount of genetic variation in the 5UTR and E2 that occurs over time is well conserved and that small variations in E2 region can occur within the same cohort but these changes occur very early in the infection. The infectivity of BVDV white deer isolates in white tailed deer was evaluated. Eight white tailed deer fawns 4-6 weeks in age were placed in 2 groups: group 1 was inoculated with non-cytopathic (NCP) BVDV type 1b, and group 2 was inoculated with NCP-BVDV type 2. A control group of two animals was also used. The two viruses were recovered in the fall of 2004 from two separate cases of unthrifty deer in South East South Dakota. The deer were negative for chronic wasting disease but were positive for BVDV by IHC and virus isolation. Virus was administered through oral and nasal routes. Clinical observations were recorded daily from -3 to 13 days post-inoculation. Average body temperatures were significantly elevated in the infected animals on day 6 for group 1 and day 7 for group 2. Average lymphocyte count for each group was <50% of control for 1 day within each group, 43% on day 3 for group 1 and 47% on day 6 for group 2. White tail deer are susceptible to BVDV infection from white tail deer isolates. It remains to be determined if deer can transmit the virus to cattle or if deer can become persistently infected as calves Georgia: To characterize management practices related to feedlot acute interstitial pneumonia (AIP), a survey was sent to managers at 561 U.S. feedlots in 21 states. Of 72 surveys returned (12.8%), 53% came from Kansas and Nebraska. The total number of cattle placed by respondents was 2,495,439, representing approximately 10% of the cattle placed in 2000. Vaccination for viral respiratory pathogens was common; fewer feedlots vaccinated for Haemophilus somnus or Pasteurella (Mannheimia). 61% of respondents practiced metaphylactic administration of antimicrobials. 65% of animals that died received a postmortem examination; 16% of respondents reported that no animals received postmortems. BRD was the leading cause of morbidity and mortality; 12.8% of placements were treated for BRD and 0.8% were died due to BRD. Of all placements, 1.3% of cattle were treated for AIP and 0.1% of placements died of AIP. Managers reported that 80.5% of AIP deaths occurred in cattle on feed over 60 days, 62% of AIP deaths occurred in the summer, and 62.2% of AIP deaths were heifers. Thirty-three feedlots reported the percent of placements that died of AIP (0.001-0.75%). Feedlots in northern states were less likely to report AIP as a cause of morbidity/mortality, while larger feedlots and feedlots placing higher proportions of yearlings more often recognized AIP as a cause of morbidity/mortality. Although heifers were recognized to account for 62% of AIP deaths, feedlots placing a large proportion of heifers were not more likely to recognize AIP as a cause of morbidity/mortality. Feedlots that vaccinated over 95% of cattle for Mannheimia/Pasteurella were less likely to recognize AIP as a cause of morbidity/mortality. NADC: ARS scientists have developed a method for isolation of respiratory tract dendritic cells from ruminant lung tissue. Respiratory tract dendritic cells were isolated using a magnetic-activated cell sorter (MACS). Following MACS purification, the cells were examined by electron microscopy and by flow cytometry for dendritic cell-specific surface molecules and tracer endocytosis. During this past year, dendritic cells have been isolated from neonatal lambs as well as adult sheep. This is the first documented isolation of dendritic cells from neonatal animals. Pulmonary dendritic cells from neonates and adults are similar phenotypically and functionally. This is in contrast to the current dogma that suggests that neonatal dendritic cells are less mature. Nebraska: BVDV has a 12.5 kb, single-stranded RNA genome consisting of a 5 untranslated region (UTR), a single large open reading frame and a 3 UTR. The 5 UTR contains an internal ribosomal entry site (IRES) necessary for translation in a cap-independent manner. Bovine viral diarrhea virus genotype 2 (BVDV2) isolates have a wide range of virulence. We have determined that in BT and BL-3 cells, the Npro coding region either had no influence or caused a significant decrease in IRES-mediated translational efficiency, with the exception of BVDV 890. Mutagenesis of IRES base 219 and/or 278 ameliorated suppression of the translational efficiency. Results suggest interactions between the Npro and nucleotides within and external to the IRES element influence translational efficiency. These results underscore the complex interplay of cellular factors influencing IRES-mediated translation. These constructs will be useful in continued studies using an infectious full-length BVDV2 clone to characterize the interplay of the IRES and the Npro coding region of BVDV2 on virulence. Vaccination against BVDV infection should protect against viremia and prevent dissemination of virus throughout the host, blocking infection of target cells. The objective of this study was to characterize the protection against infection and disease from challenge exposure with NY-1 BVDV afforded by use of a MLV noncytopathic BVDV type 1 vaccine. Vaccine virus replicated systemically in vaccinated calves as evident antemortem by decreased peripheral leukocyte and lymphocyte counts, and postmortem by lymphoid depletion in Peyers patches and mesenteric lymph nodes. Post-challenge, nonvaccinated calves developed clinical signs of disease, viremia, leukopenia, and thymic infection. In contrast, post-challenge, vaccinated calves did not exhibit fever, signs of respiratory tract disease, leukopenia, or viremia. BHV latency: We have previously demonstrated that the LR gene encodes a protein that is expressed in sensory neurons and during productive infection. Site-directed mutagenesis indicated that ORF-2 expression is required for the latency-reactivation cycle of BHV-1. The LR gene interferes with apoptosis, which promotes neuronal survival during the transition from acute infection to latency. LR-RNA sequences or a small open reading frame appears to inhibit bICP0 expression (the gene that is anti-sense to the LR gene), and consequently can inhibit viral gene expression. We believe that the LR gene encodes multiple functions that cooperate to regulate latency. The dominant function is a protein encoded by ORF-2 that is absolutely required for the latency-reactivation cycle in cattle. The LR gene also plays a role in the ability of BHV-1 to grow in the tonsils of infected calves. LR mutant virus can persist in the tonsils of latently infected calves, but it does not reactivate from latency. Studies that are in progress now to compare viral gene expression in TG or tonsils of calves latently infected with wt BHV-1 or the LR mutant. Analysis of the bICP0 gene: bICP0 is considered to be most important transcriptional regulatory gene of BHV-1. In addition to regulating transcription, bICP0 is toxic to cells. bICP0 contains a C3HC4 zinc RING finger at its amino terminus. Site-specific mutagenesis on the C3HC4 zinc RING finger revealed this domain is necessary for toxicity and eliminates transcriptional regulatory activity. Since bICP0 does not specifically bind DNA, we hypothesized that bICP0 interacts with transcription factors. Earlier studies have demonstrated that bICP0 interacts with histone deacetylase 1 (HDAC1). HDAC1 represses transcription because it removes acetyl groups from histones, thus making chromatin transcriptionally inactive. To identify bICP0 functional domains that are not part of the zinc RING finger, we developed a panel of transposon insertion mutants that span bICP0. A large domain spanning amino acids 78-256, and a separate domain that is at or near amino acid 457 was necessary for efficient trans-activation of a promoter. Sequences near the C-terminus (amino acids 607-676) contain a functional nuclear localization signal (NLS). Collectively, our studies indicated that bICP0 contains several important functional domains; 1) the zinc RING finger, 2) two separate domains that activate transcription, and 3) a C-terminal NLS that is also necessary for efficient trans-activation. Kansas: We examined the role of envelope proteins gE and US9 in the anterograde transport of BHV-1 following reactivation in TG neurons of calves. Nasal and ocular virus shedding was quantitated by plaque assay during primary infection and after reactivation. During primary infection, calves infected with BHV-1 wild type, gE deleted, or US9 deleted recombinant viruses shed similar amounts of virus from the eye and nasal cavity. When reactivation was initiated with dexamethasone, virus shedding was observed only in calves infected with wild type virus, but not in gE and US9-deleted BHV-1 viruses. The establishment of latent infection in TG neurons by these three viruses was determined by quantitating the latent genomic copy numbers by real time PCR using gB specific primer pairs and probe. The latent genomic copy numbers for all the three viruses were comparable. These results indicate that the US9 and gE-deleted BHV-1 recombinant viruses infect, grow and establish latency following infection similar to wild type virus but are not transported to the nasopharynx and eye following reactivation. gE and US9 may play a role in anterograde transport from TG neurons back to the nasal epithelium, cornea and conjunctiva. OBJECTIVE 3: Develop intervention strategies for critical control points to reduce the impact of BRD. Minnesota: We have demonstrated previously that the beta2 integrin LFA-1 is a receptor for LktA in bovine leukocytes and is involved in leukotoxin-induced biological effects. However the subunits within LFA-1 involved in binding and post-binding signaling have not been well characterized. Our research precisely pinpointed and characterized the interaction of the LFA-1 subunits with LktA. The results indicate that although LktA can efficiently bind to the CD18 subunit of both LFA-1 and Mac-1, post-binding signaling events including elevation of intracellular calcium and CD18 tail phosphorylation are only observed through LFA-1. Furthermore, LktA also binds to the CD11a subunit of LFA-1. LktA binding to CD11a could be inhibited by a small molecule inhibitor of the I- domain, the major ligand binding site on CD11a. These results suggest that CD18 subunit of bovine LFA-1 contains the LktA binding site and signaling through LFA-1 requires LktA interaction with the I-domain of CD11a, and this binding is critical in bringing about intersubunit interaction resulting in signaling. This is the first report which describes discrete functions for the CD18 and CD11a subunits in the LFA-1/LktA interaction in bovine leukocytes. Oklahoma: During the fall 2003 and winter 2004 there were reports of BHV-1 disease in cattle with prior vaccination with MLV vaccines. These episodes occurred 50 or more days after arrival/processing, and were not associated with any one vaccine. We determined if field isolates from these cases were susceptible to the BHV-1 antibodies in MLV vaccinated calves. Results of this study using 5 vaccine strains, a reference BHV-1 Colorado NVSL strain, BHV-1 Cooper challenge strain, and an isolate from a clinic case indicated a wide range of antibody titers when vaccinated calves serum were tested against these 8 strains. The titers were highest to the BHV-1 Colorado NVSL and one MLV vaccine virus (Baker strain). The lowest titers were to the BHV-1 Cooper challenge strain and the recent field strain, BHV-1-31751. In almost all sera, the differences were 3-10 fold less. Interestingly, the antibody titers in the calves serums were quite low to the homologous vaccine. It appears that most of the vaccines induced antibodies that poorly neutralized the BHV-1 Cooper challenge strain and the field strain. Wisconsin: The relationships between air quality, a variety of environmental risk factors, and calf respiratory health in 13 naturally ventilated calf barns was evaluated. A minimum of 12 pre-weaned calves were randomly selected and scored for the presence of respiratory disease at each site. An air sampling device was used to determine airborne bacteria colony forming units per cubic meter (cfu/m3) of air in calf pens and central alleys within the barns. Temperature and relative humidity were recorded in each calf pen, the barn alley, and outside the barn. Pen bedding type, dry matter, and a calf nesting score were recorded. Significant factors associated with reduced exterior wind speed and direction were determined and used to estimate building ventilation rates. Factors that were significantly associated with a reduced prevalence of respiratory disease were reduced pen log10 cfu/m3, presence of a solid barrier between each calf, different pen, and increasing nesting score. Significant factors associated with reduced alley log10 cfu/m3 were increased ventilation changes per hour, increased barn volume per kg of calf, reduced pen log10 cfu/m3, and barn type. Iowa: A survey of antimicrobial susceptibility of 223 recently recovered isolates of Mycoplasma bovis from across the U.S was completed by broth microdilution. There were no significant differences in susceptibility patterns related to geographic origin or disease presentation. Enrofloxacin, florfenicol, and spectinomycin were found to be active. Oxytetracycline and chlortetracycline were active against half the isolates; very few isolates were susceptible to tilmicosin. None were susceptible to erythromycin, ampicillin, or ceftiofur. Kansas: Needle-free and conventional needle-andsyringe injection techniques were evaluated in two studies of dairy calves and feedlot steers. Study 1: 104, 5- to 10-month old Holstein calves were vaccinated with 5-way MLV virus vaccine, Mannheimia haemolytica bacterin-toxoid and 5-way Leptospira bacterin using needle-free or conventional injection. The serological response to the IBR fraction of the 5-way viral vaccine and MH bacterin was significantly higher for the needle-free route of administration, while the serological response to the LP fraction was not significantly different. Study 2: 111 yearling feedlot steers were vaccinated with 5-way modified-live virus vaccine and Mannheimia haemolytica (MH) bacterin-toxoid, using needle-free or conventional injection. Serological response to the IBR viral fraction of the 5-way vaccine was significantly higher (P=0.001) on day 21 following needle-free injection, compared to conventional injection. Serological responses to the MH supernatant and cell-associated antigens were not statistically different. Louisiana: Research indicated that the antimicrobial peptide cecropin B is effective in inhibiting M. haemolytica colonization in the nasal passage of calves. Transfection of the upper respiratory tract with 100 ¼g of plasmid DNA per nostril inhibited colonization of a virulent strain of M. haemolytica S1 as long as 14 days after transfection. The spike glycoproteins encoded by two prototypic respiratory and enteric bovine coronaviruses were codon optimized. This codon optimization resulted in high gene expression in transient assays as compared to wild type genes. NADC: New leukotoxin mutants of M. haemolytica serotypes 1 and 6 were constructed which express and transport the C-terminal 1/3 of the leukotoxin structural gene and which may be useful for vaccination. The larger deletion introduced into the new mutants may reduce adverse reactions in Bighorn sheep. Adenoviruses appear to play a role in outbreaks of respiratory disease in domestic and wild ruminants. Additional work is necessary to characterize their role to design effective strategies for disease intervention. INTER-STATION COLLABORATIVE EFFORTS SD, NE, OK and AL collaborated on an issue of Veterinary Clinics of North America focused on bovine viral diarrhea virus MI worked with SD, KS, OK, AL, OH and NADC in identifying cattle persistently infected with BVDV for natural fetal challenge study. SD, GA, CA, and NE collaborated on BRSV challenge models SD, AL, MI, and NADC shared various BVDV strains and collaborated on developing new challenge models. SD and GA collaborated on bovine immunology studies. SD shared BHV-1 strains with KS, U of Pennsylvania and Princeton. GA collaborated with University of Montreal and NIH on new vaccine vehicles and delivery systems. GA collaborated with Merial in measuring vaccine efficacy across a multi assay platform. KS and NE shared reagents related to BHV-1 and BHV-5 research. CA shared BRSV isolates with SD and U of Tennessee. NADC and OK in constructing and evaluating new mutant P. multocida vaccine. NADC, OK and New Mexico State on evaluating mucosal immunity to oral M. haem/P. multi vaccine NADC and MN collaborated on generating leukotoxin knock out mutants of M. haemolytica. NADC, NE, OK, WI and MN share monoclonal antibodies directed against M. haemolytica leukotoxin. OK and NADC collaborated on sequencing of multiple BVDV isolates.

PEER-REVIEWED PUBLICATIONS: Rosenbusch, R. F., Kinyon, J. M., Apley, M., Funk, N. D., Smith, S., and L. J. Hoffman. In vitro antimicrobial inhibition profiles of Mycoplasma bovis isolates recovered from various regions of the United States from 2002-2003. (In Press, J. Vet Diagn. Invest.). Lu, X., and R. F. Rosenbusch. 2004. Endothelial cells from bovine pulmonary microvasculature respond to Mycoplasma bovis preferentially with signals for mononuclear cell transmigration. Molecular Pathogenesis 37: 253-261. Nanduri, B., M. L. Lawrence, and S. C. Burgess. In press. Proteomic analysis using an unfinished bacterial genome: the effects of sub-minimum inhibitory concentrations of antibiotics on Mannheimia haemolytica virulence factor expression. Proteomics. Reeks, B. Y., F. R. Champlin, D. B. Paulsen, D. W. Scruggs, and M. L. Lawrence. 2005. Effects of sub-MIC antibiotic levels and temperature on growth kinetics and outer membrane protein expression in Mannheimia haemolytica and Haemophilus somnus. Can. J. Vet. Res. 69:1-10. Boyd, B. L., Lee, T. M., Kruger, E. F., Pinchuk, L. M . 2004. Cytopathic and Noncytopathic Bovine Viral Diarrhoea Virus Biotypes affect Fluid Phase Uptake and Mannose Receptor-Mediated Endocytosis in Bovine Monocytes. Vet. Immunol. Immunopathol. 102:53-65. S.-R. Lee, G. T. Pharr, A. M. Cooksey, F. M. McCarthy, B. L. Boyd, and L. M. Pinchuk. Differential detergent fractionation for non-electrophoretic bovine blood monocyte proteomics reveals proteins involved in professional antigen presentation. In preparation. Klink H.A, R.P. Brady, C. L.Topliff, K.M. Eskridge, S. Srikumaran, C. L. Kelling. 2005. Influence of bovine respiratory syncytial virus F glycoprotein N-linked glycans on in vitro expression and on antibody responses in BALB/c mice. Vaccine. Accepted for publication. Kelling, C. L., B.D. Hunsaker, D.J. Steffen, 2005. Characterization of protection in experimentally-infected calves from systemic infection or disease by vaccination with modified-live noncytopathic bovine viral diarrhea virus type 1. American Journal of Veterinary Research. Accepted for publication. Topliff CL, Chon SK, Donis RO, Eskridge KM, Kelling CL. 2005. In vitro and in vivo translational efficiencies of 5' untranslated region from eight genotype 2 bovine viral diarrhea virus isolates. Virology 331: 349-356. Achenbach JE, Topliff CL, Vassilev VB, Donis RO, Eskridge KM, Kelling CL. 2004. Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays. Journal of Virological Methods 121: 1-6. Brady RP, Topliff CL, Kelling CL. 2004. In vitro expression of full-length and truncated bovine respiratory syncytial virus G proteins and their antibody responses in BALB/c mice. Vaccine 22: 3762-3768. Henderson, G., G.-C. Perng, A. B. Nesburn, S. L. Wechsler, and C. Jones. 2004. The latency related (LR) gene encoded by bovine herpesvirus 1 (BHV-1) can suppress caspase 3 and caspase 9 cleavage during productive infection. J. of Neurovirology. 10:64-70. Jiang, Y., M. Inman, Y. Zhang, N. A. Posadas , and C. Jones. 2004. A mutation in the latency related gene of bovine herpesvirus 1 (BHV-1) inhibits expression of proteins encoded by ORF2 and Reading Frame C during productive infection. J. of Virology 78:3184-3189. Inman, M., J. Zhou, H. Webb, and C. Jones. 2004. Identification of a novel transcript containing a small open reading frame that is expressed during latency, and is antisense to the latency related gene of bovine herpes virus 1 (BHV-1). J. of Virology 78:5438-5447. Henderson, G., Y. Zhang, M. Inman, D. Jones and C. Jones. 2004. Infected cell protein 0 encoded by bovine herpesvirus 1 can activate caspase 3 when overexpressed in transfected cells. J Gen Virol ; 85: 3511-3516 Perez, S., M. Inman, A. Doster, and C. Jones. 2005. The latency related gene encoded by bovine herpesvirus 1 (BHV-1) promotes virus growth and reactivation from latency in tonsils of infected calves. J. of Clinical Microbiology. 43: 393-401. Zhang, Y. and C. Jones. 2005. Identification of functional domains within the bICP0 protein encoded by BHV-1. J. of General Virology, 86:879-886. Geiser, V., Y. Zhang, & C. Jones. 2005. Analysis of a bovine herpesvirus 1 (BHV-1) recombinant virus that does not express the bICP0 protein. J. of General Virology. 86: 1987-1996. Confer, A.W., Fulton, R.W., Step, D.L., Johnson, B.J., Ridpath, J.F.: Viral Antigen Distribution in the Respiratory Tract of Cattle Persistently Infected with Bovine Viral Diarrhea Virus Subtype 2a. Veterinary Pathology, 42:192-199, 2005. Step, D.L, Confer, A.W., Kirkpatrick, J.G., Richards, J.B., Fulton, R.W.: Respiratory Tract Infections in Dairy Calves from Birth to Breeding Age: Detection by Laboratory Isolations and Seroconversions. Bovine Practitioner, 39:44-532, 2005. Fulton, R.W., Briggs, R.E., Ridpath, J.F., Saliki, J.T., Confer, A.W., Payton, M.E., Duff, GC, Step, D.L., Walker, D.: Transmission of Bovine Viral Diarrhea Virus 1b to Susceptible and Vaccinated Calves by Exposure to Persistently Infected Calves. Canadian Journal for Veterinary Research, 69:161-169, 2005. Welsh RD, Dye LB, Payton ME, Confer AW. Frequency of isolation and antimicrobial susceptibilities of bacterial pathogens from bovine pneumonia: 1994-2002. J Vet Diag Invest 16:426-431, 2004. Ayalew S, Confer AW, Blackwood ER. Characterization of Immunodominant and Potentially Protective Epitopes of Mannheimia haemolytica Serotype 1 Outer Membrane Lipoprotein PIpE. Infect and Immun 72:7265-7274, 2004. Prado ME, Dabo SM, Confer AW. Immunogenicity of iron-regulated outer membrane proteins of Pasteurella multocida A:3 in cattle: molecular characterization of the immunodominant heme acquisition system receptor (HasR) protein. Vet Microbiol, 105:269-280, 2005. Carter JN, Gill DR, Lalman DL, Krehbiel CR, Confer AW, Smith RA, Claypool PL, McDowell LR. Vitamin E supplementation in the diet of newly arrived feedlot cattle. J Anim Sci 83:1924-1932, 2005. Thumbikat, P., T. Dileepan, M.S. Kannan and S.K. Maheswaran. 2005. Mechanisms underlying Mannheimia haemolytica leukotoxin-induced oncosis and apoptosis of bovine leukocytes. Microbial Pathogenesis 38: 161-172. Dileepan, T., P. Thumbikat, B. Walcheck, M.S. Kannan, and S.K. Maheswaran. 2005. Recombinant expression of bovine LFA-1 and characterization of its role as the receptor for Mannheimia haemolytica leukotoxin. Microbial Pathogenesis 38: 249-257. Thumbikat, P., T. Dileepan, M.S. Kannan and S.K. Maheswaran. 2005. Characterization of Mannheimia (Pasteurella) haemolytica leukotoxin interaction with bovine alveolar macrophage b2 integrins. Veterinary Research. September Issue. Dileepan, T., M.S. Kannan, B. Walcheck, P. Thumbikat, and S.K. Maheswaran. 2005. Mapping of the binding site for Mannheimia haemolytica leukotoxin within the bovine CD18. Infection and Immunity. 73: 5233-5237. Fairbanks KK, CL Rinehart, WC Ohnesorge, MM Loughin and CCL Chase. 2004. Evaluation of fetal protection against experimental infection with type 1 and type 2 bovine viral diarrhea virus after vaccination of the dam with a bivalent modified-live virus vaccine. JAVMA 225:1898-2004 Al-Mubarak, A. and Chowdhury, S.I.. Glycoprotein I (gI)-deleted BHV-5 retains significant neurovirulence. 2004. J. Neurovirology 10:233-243. Hollis LC, Smith JF, Johnson BJ, Kapil S, Mosier DA: A Comparison of Serological Responses when Modified-live Infectious Bovine Rhinotracheitis Virus Vaccine, Mannheimia haemolytica Bacterin-Toxoid are Administered with Needle-free versus Conventional Needle-based Injection in Yearling Feedlot Steers. 2005. The Bovine Practitioner 39:106-109. Hollis LC, Smith JF, Johnson BJ, Kapil S, Mosier DA: A Comparison of Serological Responses when Modified-live Infectious Bovine Rhinotracheitis Virus Vaccine, Mannheimia haemolytica Bacterin-Toxoid and Leptospira pomona Bacterin are Administered with Needle-free versus Conventional Needle-based Injection in Holstein Dairy Calves. 2005. The Bovine Practitioner. 39:110-114. Dunn J, Kaneene JB, Grooms D, Bolin S, Bolin C, Bruning-Fann C. Effects of positive results for Mycobacterium avium subsp paratuberculosis as determined by microbial culture of feces or antibody ELISA on results of caudal fold tuberculin test and interferon-gamma assay for tuberculosis in cattle. American Journal of Veterinary Research, 2005;226:429-435 Tahir ZM, Alocilja EC, Grooms DL. Polyaniline synthesis and its biosensor application. Biosens Bioelectron. 2005;15;20(8):1690-1695. Boudreaux, C.M., Corstvet, R.E., Cooper, R.K., and Enright, F.M., 2005. Effects of cecropin B transgene expression on Mannheimia haemolytica serotype 1 colonization of the nasal mucosa of calves. A J Vet Res. In Press. Chad M Petit; Jeffery M Melancon ; Vladimir N Chouljenko; Robert C Colgrove; Michael Farzan; David M Knipe; Konstantin Kousoulas. 2005. Genetic Analysis of the SARS-Coronavirus Spike Glycoprotein Functional Domains Involved in Cell-Surface Expression and Cell-to-Cell Fusion (Virology; In press). Doyle, C.K., Labruna, M.B., Breitschwerdt, E.B., Tang, Y, Corstvet, R.E., Hergarty, B.C., Bloch, K.C., Li, P., Walker, D.H., and McBride, J.W., 2005. Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene. J Mol Diagn 7(4):1-7. Vladimir N Chouljenko, Chad M. Petit and K. G. Kousoulas. Delineation of functional domains of the spike glycoprotein encoded by enteric and respiratory strains of bovine coronavirus (In preparation). Woolums AR, Loneragan GH, Hawkins LL, Williams SM. Baseline management practices and animal health data reported by U.S. feedlots responding to a survey regarding acute interstitial pneumonia. Bov Pract, 39:116-124; 2005. Woolums AR, Loneragan GH, Hawkins LL, Williams SM. A survey of the relationshiop between management practices and risk of acute interstitial pneumonia at U.S. feedlots. Bov Pract; 39:125-133; 2005. Reber, AJ, Hippen, AR, Hurley, DJ, 2005. Ingestion of whole colostrum rapidly induces the capacity in newborn calves to stimulate and respond in one-way mixed leukocyte cultures. AJVR (in Press). Briggs, R. E., Tatum, F. M. Generation and molecular characterization of new temperature-sensitive plasmids intended for genetic engineering of the Pasteurellaceae. Appl. Environ. Microbiol. Accepted July 19, 2005. Briggs, R. E., Tatum, F. M. Construction of Pasteurella haemolytica vaccines. U.S. Patent 6,793,927 issued September 21, 2004. Fulton, R. W., Briggs, R. E., Ridpath, J. F., Saliki, J. T., Confer, A. W., Payton, M. E., Duff, G. C., Step, D. L, Walker, D. Transmission of bovine viral diarrhea virus 1b to susceptible and vaccinated calves by exposure to persistently infected calves. Can J Vet Res. 2005; 69:161-169. Meyerholz DK, Grubor B, Gallup JM, Lehmkuhl HD, Anderson RD, Lazic T, Ackermann MR. Adenovirus-mediated gene therapy enhances parainfluenza virus 3 infection in neonatal lambs. J Clin Microbiol. 2004 Oct;42(10):4780-7. Meyerholz DK, Grubor B, Fach SJ, Sacco RE, Lehmkuhl HD, Gallup JM, Ackermann MR. Reduced clearance of respiratory syncytial virus infection in a preterm lamb model. Microbes Infect. 2004 Nov;6(14):1312-9. Tatum, F. M., Yersin, Y. G., Briggs, R. E. Construction and virulence of a Pasteurella multocida FhaB2 mutant in turkeys. Microb Pathog. 2005 Jul-Aug;39(1-2):9-17. Tatum, F. M., Briggs, R. E. Construction of in-frame aroA deletion mutants of Mannheimia haemolytica, Pasteurella multocida, and Haemophilus somnus using a new temperature-sensitive plasmid. Appl. Environ. Microbiol. Accepted July 19, 2005. Atapattu, D. and C.J. Czuprynski. 2005. Mannheimia haemolytica leukotoxin induces apoptosis of bovine lymphoblastoid cells (BL-3) via a caspase-9 dependent mitochondrial pathway. Infect. Immun. (in press). Kuckleburg, C., M.J. Sylte, T. J. Inzana, L.B. Corbeil, B. Darien and C.J. Czuprynski. 2005. Bovine platelets activated by Haemophilus somnus or its lipooligosaccharide cause endothelial cell damage in vitro. Microb Pathogen. 38:23-32. Lago, A., S.M. McGuirk, T.B. Bennett,, N.B. Cook,, and K. V. Nordlund. Submitted. Calf respiratory disease and pen microenvironments in naturally ventilated calf barns in winter. J. Dairy Sci. Leite, F., D. Atapattu, C. Kuckleburg, , R. Schultz, and C.J. CZUPRYNSKI. 2004. Incubation of bovine PMNs with conditioned medium from BHV-1 infected peripheral blood mononuclear cells increases their susceptibility to Mannheimia haemolytica leukotoxin. Vet. Immunol. Immunopathol. 103:187-193. Qiu, Z., J. S. Harms, J. Zhu, and G. A. Splitter. 2004. Bovine herpesvirus tegument protein VP22 enhances thymidine kinase/Ganciclovir suicide gene therapy for neuroblastomas compared to herpes simplex virus VP22. J. Virol. 78:4224-4233. Qiu Z, Zhu J, Harms JS, Friedrichsen J, Splitter GA. 2005. Bovine herpesvirus VP22 induces apoptosis in neuroblastoma cells by upregulating the expression ratio of Bax to Bcl-2. Hum Gene Ther. 16:101-108. Slack, J., Thomas, C and Peek, S.F. 2004. Pneumothorax in dairy cattle : 30 cases (1990-2003). J. Amer. Vet. Med. Assoc. 225:732-735. Sylte, M.J., C. J. Kuckleburg, L. B. Corbeil, T. J. Inzana, P. J. Bertics, and C. J. Czuprynski. 2005. Stimulation of P2X7 enhances Haemophilus somnus lipooligosaccharide-mediated apoptosis of endothelial cells. J. Leuk. Biol. 77:958-965. Sylte, M.J. C. J. Kuckleburg, F.P. Leite, T.J. Inzana and C.J. CZUPRYNSKI. 2005. Interleukin-1 beta diminishes Haemophilus somnus lipooligosaccharide-mediated apoptosis of endothelial cells. Microb. Pathogen. (in press). Sylte, M., T. Inzana and C. J. CZUPRYNSKI. Submitted. Role of tumor necrosis factor in endothelial cell apoptosis caused by Haemophilus somnus lipooligosaccharide. Vet. Immunol. Immunopathol. Zhu J, Z. Qiu, C. Wiese, Y. Ishii, J. Friedrichsen, G. Rajashekara, and G.A. Splitter. 2005. Nuclear and mitochondrial localization signals overlap within bovine herpesvirus 1 tegument protein VP22. J. Biol. Chem. 280:16038-16044. ABSTRACTS: Nanduri, B., S. C. Burgess, and M. L. Lawrence. 2005. Sub-MIC concentrations of antibiotics decrease Mannheimia haemolytica leukotoxin expression. 105th General Meeting of the American Society for Microbiology, Atlanta, Georgia. Nanduri, B., M. L. Lawrence, and S. C. Burgess. 2005. Proteomic profiling of M. haemolytica; a bacterial pathogen with a draft genome. 53rd ASMS Conference on Mass Spectrometry, San Antonio, Texas. Lawrence, M. L. 2005. Impact of sub-MIC levels of chlortetracycline and chlortetracycline/ sulfamethazine on growth kinetics of Mannheimia haemolytica and Haemophilus somnus. Summer 2005 Academy Of Veterinary Consultants Conference, Kansas City, Missouri. Hunsaker, B.D., D.J. Steffen, CL. Topliff, C.L. Kelling, 2004. Characterization of protection in calves from systemic infection or disease by vaccination with modified-live noncytopathic bovine viral diarrhea virus type 1. Conference of Research Workers in Animal Disease. Klink,HA, R.P. Brady, C.L.Topliff, K.M.Eskridge, S. Srikumaran, C.L.Kelling. 2004. Influence of bovine respiratory syncytial virus F glycoprotein N-linked glycans on in vitro expression and on antibody responses in BALB/c mice. Conference of Research Workers in Animal Disease. Topliff, C.L., S.K.Chon, R.O. Donis, K.M. Eskridge, C.L Kelling. 2004. Translational efficiencies of the 5' untranslated region from eight genotype 2 bovine viral diarrhea virus field isolates varying in virulence. Conference of Research Workers in Animal Disease. Fulton, R.W., Burge, L.J., dOffay, J.M., Funk, R., Weaver, G.D., Van Campen, H., Johnson, B.J.: Immune Response Differences in Serums from Bovine Herpesvirus-1 Vaccinated Cattle: Depenndence of Viral Strarin. 47th Annual Meeting of AAVLD. October 22-24, 2004. Greensboro, NC. Fulton, R.W., Ridpath, J.F., Ores, S., Saliki, J.T., Burge, L.J., Confer, A.W.: Bovine Viral Diarrhea Virus (BVDV) Subtypes in Diagnostic Laboratory Accessions from Clinical and Necropsy Cases: Distribution of BVDV1a, BVDV1b, and BVDV2a Subtypes. 47th Annual Meeting of AAVLD. October 22-24, 2004. Greensboro, NC. Confer, A.W., Fulton, R.W., Step, D.L., Johnson, B.J., Ridpath, J.F.: Viral Antigen Distribution in the Respiratory Tract of Cattle Persistently Infected with Bovine Viral Diarrhea Virus Subtype 2a. 47th Annual Meeting of AAVLD. October 22-24, 2004. Greensboro, NC. Dabo SM, Anderson BE, Confer AW, Gupta S. Expression, Purification and Binding Activity of B. henselae Pap 31. Proceedings 105th Annual Meeting of American Society of Microbiology, Atlanta, GA. 2005 Ayalew S, Blackwood ER, Confer AW. Sequence diversity of the immunogenic outer membrane lipoprotein PIpE from Mannheimia haemolytica serotypes 1,2 and 6. Proceedings 105th Annual Meeting of American Society of Microbiology, Atlanta, GA 2005. Chase CCL, DM Miskimins, T Graham, L Braun, P Steen and J Ridpath. Evidence of bovine viral diarrhea virus persistent infection in two white-tail deer in southeastern South Dakota. Proceedings of the 37th annual conference of American Association of Bovine Practitioners, Fort Worth, TX, September 23-25, 2004, p. 169. Fogarty-Fairbanks K, CL Rinehart, WC Ohnesorge, MM Loughin and CCL Chase. Fetal protection against BVDV type 1 or 2 heterologous challenge. Proceedings of the 37th annual conference of American Association of Bovine Practitioners, Fort Worth, TX, September 23-25, 2004, p.171. Fogarty-Fairbanks K, J Campbell and CCL Chase. Subcutaneous dose of infectious bovine rhinotracheitis virus provides early protection against an intranasal challenge. Proceedings of the 37th annual conference of American Association of Bovine Practitioners, Fort Worth, TX, September 23-25, 2004, p. 289. Tigabu B, CCL Chase, LJ Braun. BVDV influences the expression of Toll-like receptors. Abstract W13-4. 24th Annual Meeting of American Society for Virology, University Park, PA June 18-22, 2005. Donovan, DC, Barton, MH, Norton, N, Ely, LO, Hurley, DJ. Modulation of innate immunity by in vitro treatment of keukocytes with CD14, lactoferrin and IgG, (Abstract) pg 39 Proceedings of the 84th CRWAD meeting, Chicago, IL November 14-16, 2004 Okinaga, T, Hurley, DJ, Woolums, AR, Miao, C, Zarlinga, DS. Comparison of bovine cytokine gene experession as measured by competitive and real-time PCR. (Abstract) pg 41 Proceedings of the 84th CRWAD meeting, Chicago, IL November 14-16, 2004 Okinaga, T, Hines II, ME, Hurley, DJ. Real-time PCR measurements of cytokine mRNA from cattle and goats. ( Abstract) pg 40 Proceedings of the 84th CRWAD meeting, Chicago, IL November 14-16, 2004 Reber, AJ, Okinaga, T, Tanner, M, Woolums, AR, Hurley, DJ. Development of immunity after vaccination with MLV or killed BVD vaccines. ( Abstract) pg 40 Proceedings of the 84th CRWAD meeting, Chicago, IL November 14-16, 2004 M. Wiggins, A. Woolums, D. J. Hurley, D. Cole, S. Sanchez. The prevalence of Mycoplasma bovis in a subpopulation of Georgia cattle, (Abstract) Proceedings of the 84th CRWAD meeting, Chicago, IL November 14-16, 2004 BOOK CHAPTER Vaccines, July 2005. Blackwell Publishing, Ames, IA.: Bovine Viral Diarrhea Virus: Diagnosis, Management and Control. Editors, Dr. S. Goyal and Dr. J. F. Ripdath. Pp. 209-222.