NC_OLD1192: An integrated approach to control of bovine respiratory diseases (NC-1027)

(Multistate Research Project)

Status: Inactive/Terminating

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SAES-422 Reports

Annual/Termination Reports:

[09/15/2012] [11/03/2013] [12/09/2014] [01/29/2016]

Date of Annual Report: 09/15/2012

Report Information

Annual Meeting Dates: 08/01/2012 - 08/01/2012
Period the Report Covers: 10/01/2011 - 09/01/2012

Participants

Meeting Participants (name and affiliation); * = voting member for project member station; Chris Chase* (SD), South Dakota State University; Amelia Woolums* (GA), University of Georgia; Terry Lehenbauer, University of California, Davis; Carol Chitko-McKown, USDA ARS MARC; Robert Briggs* (NADC), USDA ARS NADC ; Christine Navarre* (LA), Louisiana State University; Robert Fulton* (OK), Oklahoma State University; Derek Mosier* (KS), Kansas State University; Natalia Cernicchiaro, Kansas State University; Clinton Jones* (NE), University of Nebraska-Lincoln; Deb Hamernik (AA), University of Nebraska-Lincoln; Holly Neibergs* (WA), Washington State University; J.R. Tait, Iowa State University; Laurel Gershwin* (CA), University of California-Davis; Larry Kuehn, USDA ARS MARC; Richard Leach, USDA ARS MARC; Margo Holland, USDA NIFA; John Richeson, West Texas A&M University

Brief Summary of Minutes

Minutes

Research and outreach activities were reported by voting members in attendance and by guests from the BRD CAP (Dr. Terry Lehenbauer), USDA ARS (Drs. Carol Chitko-McKown, Larry Kuehn, and Richard Leach), Iowa State University (Dr. J.R. Tait), and WTAMU (Dr. John Richeson).

Dr. Deb Hamernik (AA) advised the project members to continue collaborating and reporting any funding leveraged from the project. She also provided a handout with information regarding various organizations working to increase funding for animal health research, with contact information (see attached handout).

Dr. Margo Holland (NIFA) provided the group with information regarding NIFA funding opportunities. She encouraged the group to apply for trainee fellowships through the NIFA Fellowship program. Some discussion ensued regarding the difficulty in funding trainees with the current limitations of the program (e.g., postdoc trainees cannot be funded until they have advanced to candidacy, by which time, at many institutions, they are nearly done with their programs). Members pointed out the difficulty in funding grad students in first 2 years of their programs&would be better if a program to do this was available. Dr. Holland encouraged members to send input, as stakeholders, to USDA. Also some discussion regarding USDA/NIH Dual Purpose Dual Benefit RFA: several felt the RFA would be more impactful if it funded research using ag animal diseases as models for human diseases, and not just research on zoonotic diseases. She encouraged the group to send feedback to Mark Mirando. She pointed out input directly from stakeholders is more impactful than her comments regarding the program. Dr. Holland also said that Foundational RFA will probably be released in late Sept., and this will have funding for FY12 and FY13 combined. She advised group to lean toward submitting if they are inclined to, as there will not be another opportunity until the FY14 RFA. She also provided other information regarding funding opportunities, FY11 funding rates, and personnel changes at NIFA (see attached handout).

Discussion of potential BRD Symposium 2014. Dr. Amelia Woolums led discussion of whether group should hold a 2014 BRD Symposium, which would be 5 years after 2009 Symposium (BRDS 2009). Dr. Woolums gave group a short summary of 2009 meeting, noted that BRDS 2009 website is still active (www.brdsyomposium.org), and that people can still access papers and powerpoints from the meeting on the site. Also, the meeting made $19,000 in profit; the money has been used to provide an additional $1000 for each of 3 student research awards at AABP, and $10,000 is available for seed money for next Symposium if group decides to hold one. Consensus was that BRDS 2014 was a good idea. Should be held in conjunction with BRD CAP if they are willing; Dr. Woolums will contact Dr. Alison Van Eenennaam (extension leader for CAP) to see if her group is willing to work together on this. Dr. Fulton suggested we try to get more involvement from AAVLD and from producer organizations (NCBA, Heifer Rearers Organization, etc.). Volunteers to serve on BRDS 2014 Organizing Committee: Robert Fulton, Christine Navarre, Carol Chitko-McKown, Laurel Gershwin, Amelia Woolums, Chris Chase. Plan to add Dr. Van Eenennaam; Dr. Dale Grotelueschen (Pfizer) was also recommended as a good candidate to join committee as rep from industry. Proposed site: immediately before 2014 AVC Summer meeting (location TBA); consensus was that AVC was easy to work with and very helpful in terms of local logistics. Amelia Woolums or Chris Chase will contact AVC leadership to ask if we can hold BRDS 2014 in conjunction with 2014 AVC summer meeting.

Business meeting: election of new Chair and Secretary. Dr. Chase (Chair) and Dr. Woolums (Secretary) have been serving for several consecutive years. They asked group if the group wished to elect new officers; they are willing to continue serving but also would be very happy for new people to step in. Group felt they were doing a good job and asked that they continue serving. Vote was taken with unanimous agreement for Dr. Chase to stay on as Chair and Dr. Woolums to stay on as secretary.

Discussion of 2013 meeting: decided to meet in conjunction with AABP, since tentative plan for 2014 will be to meet in conjunction with Summer AVC meeting. Date for 2013 NC1192 Technical Committee meeting: September 18, 2013, Milwaukee WI (in conjunction with AABP). More details and info re hotel will be sent out in early summer 2013.

Accomplishments

Accomplishments Objective 1: To aid the rapid identification and subsequent management of BRD by developing, validating and guiding the application of new state-of-the-art diagnostic tools. KS, WI, and SD reported results from state diagnostic lab testing for BRD pathogens. MI continued work to adapt a human RSV diagnostic test for rapid patient-side diagnosis of BRSV, with testing of clinical samples and comparison of results of the HRSV test with RT-PCR. The HRSV diagnostic assay was in agreement with RT-PCR 50 of 66 times (k=0.484). There appears to be a problem with false positive results (assuming the rtPCR is correct). This could be a matrix problem; MI is currently working to understand this phenomenon. NE is undertaking studies to examine the viruses that are present in calves suffering from BRDC versus healthy calves, based on the hypothesis that there novel viruses present in the respiratory tract of healthy cattle versus those suffering from BRDC. Nasal swabs from health calves will be compared to those with BRD. PCR and the ViroChip will be used to identify known viruses associated with BRDC. Deep sequencing will also be performed to identify novel viruses that are present. If new viruses are discovered, long-term studies will focus on growing these viruses and testing whether they cause disease in cattle. Sampling of herds with a history of BRD is underway. OK in collaboration with NADC isolated BVDV from 1264 PI cattle entering a feedlot between 2004 2008. Of the 1264 PI isolates there were 12.0% BVDV 1a, 78.3% BVDV1b, and 9.7% BVDV2a. The BVDV1b was the predominant subtype (P value <0.05). VA continued work to develop a highly sensitive and mobile detection device that will identify the presence of H. somni among livestock. The detector under study uses nanoparticle-based optical fiber biosensors (NOFS) to identify H. somni in materials such as nasal secretions from infected cattle. The sensor uses oligonucleotide sequences specific to different H. somni isolates coupled with an ionic self-assembled multilayer (ISAM) films onto the surface of the optical fiber cladding of the biosensor. The ISAM/probe duplex hybridized with the target DNA, and was detected and quantified based on the alteration of optical power transmitting through the fiber. Hybridization of the probe with DNA derived from the sample results in a significant decrease in the optical power transmitted through the fiber. The resulting portable sensing method would be useful for field applications where compact equipment (smaller than a laptop) can be combined to simply include a light source, optical fiber, detector, and computer. During specimen testing, DNA hybridization will alter the optical properties of the attached thin film, which will immediately modify the transmission characteristics of the fiber and produce an observable output indicating the presence and concentration of each target antigen. Specificity is provided by the careful selection of DNA probes. Sensitivity is obtained by tailoring the optical fiber and thin-film fabrication process and refining the signal processing algorithm. Objective 2: To elucidate key steps in the dynamic interactions between pathogens, host immunity and the environment, and to determine how manipulation of these factors can reduce the risk of BRD CA has collaborated with WA, SD, OK, and the BRD CAP to carry out challenge studies using individual BRD pathogens, for the purposes of characterizing host gene expression in response to infection with individual pathogens. NC1192 input to CAP during 2011 meeting led CAP to increase the number of cattle per group (from n = 2 to n = 6), which all agreed would improve the strength of the research findings. KS evaluated risk factors associated with feedlot BRD incidence and found that factors related to weather, as well as characteristics related to the cohort (month and year of arrival, gender, days on feed, risk designation, and other factors) and distance transported were significantly associated with feedlot BRD. WI continued to investigate extracellular trap formation by bovine leukocytes exposed to M. haemolytica or its leukotoxin (LKT). Bovine monocyte-derived macrophages and alveolar macrophages, but not freshly isolated peripheral blood monocytes, produce extracellular traps (which we term METs) upon exposure to M. haemolytica or its LKT in vitro. Some of the M. haemolytica cells are ensnared within METs and a portion of those cells are killed within METs. Histophilus somnii cells also elicit trap formation by bovine leukocytes in vitro in a dose- and time-dependent manner. WI performed gene microarray analysis of bovine bronchial epithelial cells exposed in vitro to BHV-1, M. haemolytica, or both agents. Preliminary analysis revealed differential regulation (>2 fold, P<0.05) of 978 transcripts by BHV-1 alone, 2040 transcripts by M. haemolytica alone, and 3500 genes by BHV-1 and M. haemolytica in combination. Functional analysis of the microarray data revealed alterations in genes involved in biological processes of cell proliferation, inflammation, cell death, leukocyte migration, and cell surface markers. NE continued research to determine how the BHV-1 LR gene regulates latency and how a viral transcriptional activator (bICP0) stimulates productive while repressing innate immune responses. It was found that the LR gene encodes two micro-RNAs that are expressed during latency, and that both micro-RNAs interact with the RNA sensor, RIG-I, which stimulates the IFN-b signaling pathway. In collaboration with LA, NE showed that bICP27, a viral early protein that shuttles between the nucleus and cytoplasm, inhibits the transcriptional activity of two bovine IFN-² gene promoters (IFN-²1 and IFN-²3) in a dose dependent fashion. These studies provided evidence that bICP27 inhibited IFN-²1 and IFN-b3 promoter activity, thus interfering with the host response to BHV-1 infection. OK and NADC collaborated to evaluate an outbreak of severe BVDV-induced disease occurring in 2 lots of cattle entering a Texas feedlot. Affected animals had severe mucosal lesions in the oral cavity, larynx, and esophagus. Mucosal lesions varied from small (13 mm) infrequent mucosal ulcerations to large (5 mm to 1 cm) and coalescing ulcerations. A calf persistently infected with BVDV arrived with one lot and the isolated virus was genotyped as BVDV1b. Identical BVDV1b strains were isolated from 2 other mortalities. A BVDV2a genotype was also isolated in this outbreak. This genotype was identical to all BVDV2a strains isolated in both lots. OK and NADC assessed the genetic variability of M. haemolytica isolates obtained from fatal BRD cases in the United States (USA) and Australia using REP-PCR and sequencing from the 16s ribosomal sequence. All characterization methods were capable of discriminating between isolates. Modest to moderate diversity was seen amongst the isolates, with as much variation being present within a continent as between the two. This suggests that utilizing samples from diverse origins should permit extrapolation to isolates with distant geographic and temporal relationships. It also suggests that measures effective against the bacterium in one setting can reasonably be expected to be efficacious in another. Further, this information can serve as a baseline in assessing whether M. haemolytica truly is an opportunistic pathogen, or if there are notable features that distinguish commensal isolates from those more likely to be associated with disease. SD and CO collaborated to evaluate the effect of BVDV on innate immunity in the developing fetus. Liver samples were collected at necropsy from fetuses retrieved by cesarean section at gestational day 89, from artificially inseminated heifers. Four of the eight heifers had been inoculated at 75 days in gestation with a type 2 non-cytopathic BVDV strain (96B2222). BVDV antigen was clearly detected in persistently infected tissues in cells primarily located in liver sinusoids and near central veins. Confocal microscopy positively identified the population of cells infected with BVDV in fetal livers as exclusively Kupffer cells. Kupffer cells were not uniformly infected with BVDV at this stage of infection; as uninfected Kupffer cells were also observed in all tissues tested. Kupffer cells are responsible for presenting antigen to lymphocytes that were also present at this critical stage of gestation. In the context of the specific microenvironment of the liver, antigen presentation likely results in systemic tolerance rather than immune activation, contributing to persistent infection. WA serves as the repository for the collection, DNA extraction and storage of 2033 Holstein calf samples (cases/controls) from California and 795 Holstein heifer calf samples from New Mexico as part of the BRD consortium formed from the Agriculture and Food Research Initiative. US Department of Agriculture project 2011-68004-30367 Integrated program for reducing bovine respiratory disease complex in beef and dairy cattle. All of these samples have diagnostic testing for Histophilus somni, Pasturella multicoda, Mannheimia haemolytica, Mycoplasm spp, Arcanobacterium pyrogenes, bovine corona virus, bovine respiratory synctial virus, bovine viral diarrhea virus, and interstitial bovine respiratory virus completed. WA also continues work to identify loci in cattle associated with susceptibility to BRD pathogens in general and also individual pathogens in beef and dairy cattle is ongoing. A feedlot study is beginning this year that will add to 1000 beef cattle to the animal resource populations that the BRD Consortium has been developing. Objective 3: To investigate the mechanisms by which infectious agents work singly or in combination to evade, suppress, or misdirect the host immune response, or to directly induce cellular or molecular pathology, in BRD. CA has evaluated the response of bovine respiratory epithelium to exposure to BRSV and a culture supernatant of H. somni, using microarray analysis of RNA from exposed cells and ELISA. Genes expressed include IL-8, IL-6, prostaglandin synthase, and several matrix metalloproteinases. Results showed good correlation between the results of the microarray and the ELISA, indicating that the microarray accurately predicted expression of the proteins which mediate the inflammatory response to infection. GA recently completed a pilot study to evaluate the impact of gastrointestinal nematode (GIN) parasitism on immune responses to vaccination. In a small study using nursing beef calves, calves with moderate levels of GIN had lower concentrations of SN antibody to BHV-2 at 45 days post weaning and vaccination with a 5-way viral vaccine, as compared to calves with low parasite burdens. Calves with low parasite burdens showed a trend toward increased expression of IFN gamma in response to exposure to BVDV. MI continued work to test a regional BVDV eradication program in the Upper Peninsula (UP) of Michigan. Thru December 31, 2011, 294 (out of an estimated 500 herds in the UP) herds have signed up for the program. In the first five counties, 80% of herds have agreed to participate. Testing has occurred in 232 herds and BVDV PIs have been confirmed in 9 herds (3.9%). Of 17, 917 cattle screened, 24 have been confirmed as PIs (0.13%). One stakeholder biosecurity practice started has been the mandatory BVDV testing of cattle participating in the UP State Fair. SD continued research of the effect of BVDV on dendritic cell antigen presentation. Monocytes were isolated from Holstein Friesians (H.F.) and Brown Swiss (B. Swiss) calves that were 8 months to 1 year of age. Monocytes were differentiated into MDDCs using bovine recombinant IL-4 and GMCSF and confirmed morphologically and phenotypically to be MDDC. The MDDCs had long dendrites and were 5-7 times larger size then monocytes. The cell surface phenotype was CD14-, CD21-, MHCI+, MHCII+, CD86+, DEC205+. 100% of the B. Swiss calves produced MDDCs while only 5.5% of the H.F. were able to generate MDDCs. For MDDCs infection, 4 strains of BVDV were used. No infectious virus production by MDDCs occurred. Interestingly, viral RNA increased in MDDCs through 144 hr after infection. The kinetics of viral RNA production along with the amount of viral RNA was significantly different between viral stains. The study revealed that BVDV replicates in MDDCs but does not produce infectious particles. Accumulation of viral RNA may have significant effects on immune response mounted by MDDCs. VA has continued work to characterize the role of the H. somni biofilm in resistance to host defenses in vitro and in vivo. Following experimental challenge of calves it became clear that Pasteurella multocida, and possibly other organisms, cohabitated with H. somni in biofilms. Therefore, polymicrobial biofilm formation was studied using a drip flow bioreactor (DFR), which can more closely simulate biofilm formation in the lung. A laboratory biofilm was established with H. somni 2336, M. haemolytica A1, and P. multocida A:3. H. somni, M. haemolytica, P. multocida biofilms grown in the DFR resulted in mean viable biofilm log densities of 9.08, 6.9 and 9.56, respectively. When all species were grown together the mean viable biofilm log density was 9.61. In the mixed biofilm, M. haemolytica was not present, and the mean viable biofilm log densities of H. somni and P. multocida were 8.8 and 9.46, respectively. Scanning electron microscopy (SEM) of a mixed biofilm grown in the DFR showed that P. multocida and H. somni co-existed in the same biofilm. Objective 4: To develop management practices, including rationally applied therapeutic and preventative interventions that minimize the impact of BRD on cattle health, welfare and productivity CA is working to develop new vaccine candidates for co-administration to protect against BRSV and H. somni. The selected immunogen for BRSV has been cloned and expressed, and upscale expression for vaccine production is underway. The IbpADR2 from H. somni has been expressed and purified. In a separate study, IbpADR2 has been expressed in microalgae to provide a particulate recombinant antigen for vaccination of calves. GA, KS, NE, and collaborators at 3 other institutions have conducted a survey of cow-calf producers to determine herd-level risk factors for nursing (preweaning) beef calf respiratory disease. Results indicated that certain operation characteristics or management practices were significantly associated either with occurrence of calf BRD or cumulative calf treatment incidence. KS continued work to investigate clinical, behavioral, and pathophysiological data of potential value in detecting the onset and progression of BRD using experimental Mannheimia haemolytica and Mycoplasma bovis pneumonia models. Clinical illness scores (CIS) were assessed in calves challenged with M. bovis and remote triangulation devices recorded the amount of time calves spent in specific areas (near feed bunk, water, and shelter). Calves with more severe disease traveled less distance and spent less time at the hay bunk while spending more time in the shelter. The distance calves traveled was associated with the amount of lung lesion. LA developed a subunit vaccine that can elicit strong antibody response against respiratory bovine coronavirus in immunized animals. This method utilized an initial DNA vaccine encoding either the soluble portion of the spike glycoprotein or the soluble portion of the spike glycoprotein fused in-frame to bovine CD154. Animals responded to vaccination, and fusion of CD154 to the soluble portion of the spike glycoprotein resulted in a pronounced increase in circulating and neutralizing serum antibody specific for the BRCoV spike glycoprotein. LA also developed and tested a vaccine from an envelope protein mutant of bovine herpesvirus type 1 (BHV-1). This vaccine induced higher cytotoxic T cell activity and higher serum neutralizing titers than wild type vaccine. Vesicles formation is a mechanism bacteria use for specific secretion and transfer of macromolecules such as proteins and toxins to animals. OK conducted research to determine whether components of vesicles of M. haemolytica were immunogenic in mice and cattle. LC-MS/MS spectrometric analysis of vesicles from this bacterium identified 226 proteins out of which 104 (46%) were cytoplasmic, 5 (2.2%) cytoplasmic membrane, 1 (0.44%) extracellular, 58 (25.6%) OMPs and periplasmic and 58 (25.6%) unknown. Vesicles from M. haemolytica were used to vaccinate dairy calves and Balb/c mice. Analysis of sera from calves and mice by ELISA showed that circulating antibodies against M. haemolytica whole cells and leukotoxin were significantly higher on day 21 and 28 (p < 0.05) compared to day 0. Lesion scores of lungs from vaccinated calves (15.95%) were significantly (p < 0.05) lower than those from non-vaccinated calves (42.65%). Sera from mice on day 28 and calves on day 21 showed that there was 100% serum bactericidal activity in the presence of complement, while there was no killing activity in control mice calves sera. SD along with CO and investigators in Texas, Illinois, Missouri, and New York are involved in the Genetics of Feedlot Health Project which was performed in 2009 and 2010. This study looked at the impact of behavior, genetics, and nutrition, along with microbiology and immunology, as related to respiratory disease and carcass quality. Analysis of the data is continuing, including assessment of cortisol and its relationship to antibody production and proinflammatory responses Objective 5: To promote open scientific exchange and dialogue among scientists, veterinarians, allied industry professionals and cattlemen to advance BRD research initiatives. Members from WA, CA, and SD are participants in the current USDA BRD Coordinated Agricultural Project (CAP), which is currently focused on research to determine the influence of genetics in BRD. Members from other stations (MI and GA) have advised the CAP in their research and outreach efforts. Members from all participating stations presented abstracts and lectures to scientists, veterinarians, allied industry professionals, and cattlemen at various conferences in the past year. Objective 6: To facilitate the translation of research findings to practical field application by developing and integrating BRD educational programming for national veterinary and producer organizations focused on cattle health and management SD, MI, and GA have communicated with the BRD CAP re planning outreach activities related to BRD prevention and control. A presentation was made at the NCBA Cattlemens College on BRD in Nashville TN in February 2012. Also classes specific on bovine respiratory disease were taught to the advanced group of the Southern Great Plains Dairy Consortium-Teaching at Clovis NM in June 2012 KS is the home of the Beef Cattle Institute (BCI). The purpose of the Beef Cattle Institute is to create a collaborative environment to tackle issues facing the beef industry through education, research and outreach. The institute enhances the education and the value of the degrees of students, increases information access and training opportunities for people working in the beef industry, and improves the cultural and intellectual diversity of our student body. The institute sponsors graduate and undergraduate certificate programs, workshops and seminars, and beef cattle performance and health training. Sponsors and partners include AVMA, KVMA, KLA, Kansas Farm Bureau, AVC, AABP, and many industry partners (Bayer, Merial, Pfizer, Novartis, Boehringer Ingelheim). The diagnosis of bovine respiratory diseases (BRD) poses significant challenges to the clinician as there are numerous infectious etiologies, operating singly or most often in combination. OK investigators recently published a review article that describes the traditional tests and several molecular tests and discusses the benefits and limitations of the tests and their interpretation. This article makes the important point that clinicians should consult with their diagnostic laboratory regarding interpretation of test results. The rate of development and use of molecular diagnostic tests have outpaced validation, standardization, and standards for interpretation.

Publications

Maunsell FP, Woolums AR, Francoz D, Rosenbusch RF, Step DL, Wilson DJ, Janzen ED. Mycoplasma bovis infections in cattle. J Vet Intern Med 2011; 25:772-783. Woolums AR, Berghaus RD, Berghaus LJ, Ellis RW, Pence ME, Saliki JT, Hurley KAE, Galland KL, Burdett WW, Nordstrom ST, Hurley DJ. The impact of route and timing of multivalent respiratory vaccination on immune responses to booster vaccination at weaning in calves. Am J Vet Res 2012, accepted. R.L. Larson and D.L. Step. Evidence-based effectiveness of vaccination against Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni in feedlot cattle for mitigating the incidence and effect of bovine respiratory disease complex. In: Buczinski S. and Vandeweerd J., ed. Veterinary Clinics of North America: Food Animal Practice Evidence-based veterinary medicine for the bovine veterinarian. Vol 28(1), Philadelphia, PA: W.B. Saunders Company; 2012: 97-106. N. Cernicchiaro, D.G. Renter, B.J. White, J.T. Fox. Associations between weather conditions during the first 45 days following feedlot arrival and daily respiratory disease risks in autumn-placed U.S. feeder cattle. J Anim Sci. 2012 Apr 90(4): 1328-37. PMID:22147846 B.J. White, D.E. Anderson, D.G. Renter, R.L. Larson, D. Mosier, L. Kelly, M. Theurer, B. Robért, M. Walz. Clinical, behavioral, and pulmonary changes following Mycoplasma bovis challenge in calves. Am J Vet Res April 2012 73(4): 490-497. PMID: 22452495 Cernicchiaro, N., Renter, D.G., White, B.J., Babcock, A.H., Fox, J.T. Associations between weather conditions during the first 45 days following feedlot arrival and daily respiratory disease risks in autumn-placed U.S. feeder cattle. J Anim Sci. 2012; 90(4):1328-1337. Cernicchiaro, N., White, B.J., Renter, D.G., Babcock, A.H., Kelly, L., Slattery, R. Associations between the distance traveled from sale barns to commercial feedlots in the United States and overall performance, risk of respiratory disease, and cumulative mortality in feeder cattle during 1997 to 2009. J Anim Sci. 2012; 90(6):1929-1939. Cernicchiaro, N., White, B.J., Renter, D.G., Babcock, A.H., Kelly, L., Slattery, R. Effects of weight loss during transit from sale barns to commercial feedlots on health and performance in feeder cattle cohorts arriving to feedlots from 2000 to 2008. J Anim Sci. 2012; 90(6):1940-1947. White BJ, Anderson D, Renter DG, Larson R, Mosier D, Kelly L, Theurer M, Robért B, Walz M. Clinical, behavioral, and pulmonary changes following Mycoplasma bovis challenge in calves. Am J Vet Res. 2012; 73(4):490-497. Hanzlicek G, Renter DG, White BJ, Wagner BA, Dargatz DA, Sanderson MW, Scott HM, Larson RE. Management practices associated with the rate of pre-weaning calf respiratory disease from a national survey of U.S. cow-calf operations. J Am Vet Med Assoc. In press. Babcock AH, White BJ, Renter DG, Dubnicka S, Scott HM. Predicting cumulative risk of bovine respiratory disease complex using feedlot arrival data and daily morbidity and mortality counts. Can J Vet Res. In press. Wei H., Wang, Y and Chowdhury, S.I.(2011). Bovine herpesvirus type 1 (BHV-1) UL49.5 luminal domain residues 30-32 are critical for MHC-I down-regulation in virus-infected cells. PLoS one 6(10) e25742. Wei H., He J., Paulsen D.B., Chowdhury, S.I. (2012) Bovine herpesvirus type 1 (BHV-1) UL49.5 luminal domain residues 30-32 and cytoplasmic tail residues 80-96 induce better immune responses in claves than the wild-type strain Cooper. Veterinary Immunol. Immunopathol. 147: 223-229. Lum, B, V. N. Chouljenko, and K. G. Kousoulas. A subunit vaccine consisting of the respiratory bovine coronavirus (RBCoV) spike glycoprotein fused-in frame with the bovine CD40 ligand generates high titer neutralizing antibody against RBCoV. Submitted. Corbett EM, Grooms DL, Bolin SR. Evaluation of Skin Samples by RT-PCR Following Immunization with a Modified-Live Bovine Viral Diarrhea Virus Vaccine. Am J Vet Res. 2012;73(2):319-324. Aulik, N., K. Hellenbrand, D. Kisiela, and C. Czuprynski. 2011. Mannheimia haemolytica leukotoxin binds cyclophilin D on bovine neutrophil mitochondria that is not inhibited by cyclosporine A. Microb. Pathogen. 50:168-178. Aulik, N.A., K. M. Hellenbrand, and C. J. Czuprynski. 2012. Mannheimia haemolytica and its leukotoxin cause macrophage extracellular trap (MET) formation by bovine macrophages. Infect. Immun. 80:1923-1933 Da Silva, L.F. and C. Jones. 2012. Two micro-RNAs encoded within the BHV-1 latency related (LR) gene promote cell survival by interacting with RIG-I and stimulating nuclear factor-kappa B (NF-kB) dependent transcription and beta-interferon signaling pathways. J Virol, 86:1670-1682. Workman, A., J. Eudy, L. Smith, L. Frizzo da Silva, D. Sinani, H. Bricker, E. Cook, A. Doster, and C. Jones. 2012. Cellular transcription factors induced in trigeminal ganglia during dexamethasone-induced reactivation from latency stimulate bovine herpesvirus 1 productive infection and certain viral promoters. J Virol 86: 2459-2473. Frizzo da Silva, L. and C. Jones. 2012. The ICP27 protein encoded by bovine herpesvirus type 1 (bICP27) interferes with promoter activity of the bovine genes encoding beta interferon 1 (IFN-²1) and IFN-²3. Virus Research, In Press. Pittayakhajonwut, D., D. Sinani, and C. Jones. 2012. A protein (ORF2) encoded by the latency related gene of bovine herpesvirus 1 interacts with DNA. Submitted to Journal of Virology. Holland, B.P., Step, D.L., Burciaga-Robles, L.O., Fulton, R.W., Confer, A.W., Rose, T.K., Laidig, L.E., Richards, C.J., Krehbiel, C.R.: Effectiveness of Sorting Calves With High Risk of Developing Bovine Respiratory Disease on the Basis of Serum Haptoglobin Concentration at the Time of Arrival at a Feedlot. American Journal for Veterinary Research, 72: 1349-1360, 2011. Hessman, B.W, Sjeklocha, D.B., Fulton, R.W., Ridpath, J.F., Johnson, B.J., McElroy, D.R.: Acute Bovine Viral Diarrhea Associated with Extensive Mucosal Lesions, High Morbidity and Mortality in a Commercial Feedlot. Journal for Veterinary Disease Investigation, 24: 397-404, 2012. Fulton, R.W, Confer, A.W.: Laboratory Test Descriptions for Bovine Respiratory Disease Diagnosis and Their Strengths and Weaknesses: Gold Standards for Diagnosis, Do They Exist? Canadian Veterinary Journal, 53:754-761,2012 Ayalew S, Shrestha B, Montelongo M, Confer AW. Identification and immunogenicity of Mannheimia haemolytica S1 outer membrane lipoprotein PlpF. Vaccine 47: 8712-8718, 2011 Ayalew S, Shrestha B, Montelongo M, Wilson AE, Confer AW. Immunogenicity of Mannheimia haemolytica recombinant outer membrane proteins SSA-1, OmpA, OmpP2, and OmpD15. Clin & Vaccine Immunol 18: 2067-2074, 2011 Ayalew S, Shrestha B, Payton ME, Confer AW. A Rapid Microtiter Plate Serum Bactericidal Assay Method for Determining Serum Complement-mediated Killing of Mannheimia haemolytica. J Microbiological Methods 89: 99-101, 2012 Singh K, Confer AW, Step DL, Rizzi T, Wyckoff JH, Weng HY, Ritchey JW. Cytokine expression by pulmonary leukocytes from calves challenged with wild-type and leukotoxin-deficient Mannheimia haemolytica. The Vet J 192: 112-119, 2012 Stevens ET, DU Thomson, BW Wileman, S ODell, CCL. Chase. 2011 The Survival of Bovine Viral Diarrhea Virus on Materials Associated with Livestock Production. The Bovine Practitioner 45:118-123. Ridpath JF, JD Neill, CCL Chase. 2012. Impact of BVDV Infection of White-Tailed Deer During Second and Third Trimesters of Pregnancy. J Wild Dis 48:758-762. Sandal, I., T.J. Inzana, A. Molinaro, C. De Castro, J.Q. Shao, M.A. Apicella, A.D. Cox, F. St. Michael, and G. Berg. 2011. Identification, structure, and characterization of an exopolysaccharide produced by Histophilus somni during biofilm formation. BMC Microbiol. 11:186. Siddaramappa, S., J.F. Challacombe, A.J. Duncan, A.F. Gillaspy, M. Carson, J. Gipson, J. Orvis, J. Zaitshik, G. Barnes, D. Bruce, O. Chertkov, J.C. Detter, C.S. Han, R. Tapia, L.S. Thompson, D.W. Dyer, and T.J. Inzana. 2011. Horizontal gene transfer in Histophilus somni and its role in the evolution of pathogenic strain 2336, as determined by comparative genomic analyses. BMC Genomics. 12:570. Elswaifi, S.F., W.K. Scarratt, T.J. Inzana. 2012. The role of lipooligosaccharide phosphorylcholine in colonization and pathogenesis of Histophilus somni in cattle. Vet. Res. 43:49. Inzana, T.J., R. Balyan, and M. D. Howard. 2012. Decoration of Lipooligosaccharide with N-acetyl-5-neuraminic acid contributes to Histophilus somni virulence and resistance to host defenses. Vet. Microbiol. In press. Zanella, R., E.G. Casas, G.D. Snowder, H.L. Neibergs. 2011. Fine mapping of loci on BTA2 and BTA26 associated with bovine viral diarrhea persistent infection and linked with bovine respiratory disease in cattle. Frontiers in Livestock Genomics. 2:82. doi: 10.3389/fgene.2011.00082 Neibergs, H.L., R. Zanella, E. Casas, G.D. Snowder, J. Wenz, J.S. Neibergs, D. Moore. Loci on BTA2 and BTA26 are linked with bovine respiratory disease and associated with persistent infection of bovine viral diarrhea virus. 2011. Journal of Animal Science 89:907-915.

Impact Statements

Studies to evaluate host gene transcription in response to infection with BRD agents will improve understanding of the protective response to these infectious agents, forming the basis for future development of vaccines or immune stimulants which activate these protective mechanisms and improve resistance of cattle to BRD.
The studies to evaluate the impact of nematode parasitism on the response to viral vaccines will form the basis for development of preventative treatment protocols that veterinarians and producers can use to improve the response of calves to vaccination.
The study of risk factors for nursing calf respiratory disease has the potential to provide veterinarians and producers with the information they need to develop better methods of management to prevent calf respiratory disease.
Efforts to bring new information to veterinarians and producers regarding the causes and prevention of BRD should lead to improvements in the health and productivity of cattle.
Defining those conditions for specific cattle populations that enhance BRD may enable cattle health managers to develop models that predict and potentially manage these effects more effectively.
Use of behavior monitoring systems may aid in recognition of respiratory tract disease in calves. Changes in any of the parameters evaluated could lead to enhanced diagnostic and prognostic methods so that disease can be recognized sooner and treatments initiated earlier in the disease process.
Engineered vaccines against respiratory bovine coronavirus and BHV-1 should be safer, less expensive, and more effective than traditional vaccines.
The adaptation of the HRSV test for rapid detection of BRSV patient side would facilitate timely disease diagnosis and the institution of appropriate therapies and control plans.
Studies of macrophage and neutrophil extracellular traps are clarifying a relatively newly recognized mechanism of host defense, providing new knowledge regarding a basic mechanism of host defense.
Research to characterize -1 latency and suppression of host response will lead to the development of new MLV vaccines engineered to induce protective immunity without inducing latency; this will protect cattle from disease which could occur if vaccine-related latent BHV-1 is later reactivated.
Research to detect novel viruses present in healthy calves but not calves with BRD will provide new information regarding the microbial ecology of the bovine respiratory tract, and will form the basis for future research to protect calves from BRD by manipulation of the respiratory microbial population.
Characterization of BVDV isolates from PI cattle will help vaccine manufacturers understand what subtypes of the virus need to be included in vaccines in order to protect cattle against infection and generation of PI cattle.
The identification, cloning, and production of subunit components of M. haemolytica and P. multocida offer opportunity for new bacterial vaccines to better control BRD.
Guidance to veterinarians regarding interpretation of molecular and traditional diagnostic tests should improve the ability of veterinarians to make accurate and informed decisions regarding the etiology of BRD in outbreaks they manage.
The studies of infection of fetal Kuppfer cells are providing new foundational information regarding mechanisms of pathogenesis of persistent infection by BVDV. This not only improves knowledge regarding BVDV-induced disease, but improves basic understanding of development of the fetal immune response.
Research to assess the response of dendritic cells to BVDV infection will improve basic understanding of how viruses evade and impair the immune response; this information will form the basis for development of mechanisms to counteract virus-induced immune suppression in cattle and other species.
The development of optical sensors to identify H. somni should lead to chute-side diagnostic tests, allowing rapid identification of specific agents and syndromes, and providing veterinarians with data to support timely and informed decision making regarding treatment and management of BRD.
In vitro studies of biofilms containing H. somni, P. multocida, and M. haemolytica will improve foundational knowledge regarding the means by which these bacteria establish in the host and evade host immunity, which should lead to identification of novel targets for intervention to treat or prevent infection by these agents.
Work to evaluate genetic loci associated with BRD susceptibility may allow producers to select cattle with improved resistance to BRD, providing an additional management tool to limit the impact of BRD in U.S. cattle.

Date of Annual Report: 11/03/2013

Report Information

Annual Meeting Dates: 09/18/2013 - 09/18/2013
Period the Report Covers: 10/01/2012 - 09/01/2013

Participants

John Baker (AA), Michigan State University; Chris Chase*, South Dakota State University; Carol Chitko-McKown, USDA MARC; Chuck Czuprynski*, University of Wisconsin, Madison; Jose Rivera, University of Wisconsin, Madison; Ismail Boukahil, University of Wisconsin, Madison; Danielle Doyle, University of Georgia; Laurel Gershwin*, University of California, Davis; Dan Grooms*, Michigan State University; Dale Grotelueschen, University of Nebraska-Lincoln; Tom Inzana*, Virginia Tech; Terry Lehenbauer, University of California, Davis; Christine Navarre*, Louisiana State University; Maria Prado, University of Tennessee; John Richeson, West Texas A&M University; Brian Vander Ley*, University of Missouri;Brad White, Kansas State University; Amelia Woolums*, University of Georgia.
*voting member from their station

Brief Summary of Minutes

Meeting date and time: Wednesday September 18, 2013; 8 AM 5 PM
Location: Mitchell Room of the Hilton Milwaukee City Center, Milwaukee WI

Wednesday September 18, 2013

Meeting Agenda:

8:00 AM: Registration
8:30 AM: Welcome
8:40 AM 10 AM: Station reports
10 AM 10:15 AM: Break
10:15 AM 12:00 PM: Station reports
12 1 PM: Lunch (provided at meeting)
1-1:30 PM: Update from NC1192 Advisor John Baker
1:30 2:00 PM: Update from the BRD CAP (Laurel Gershwin and Terry Lehenbauer)
2:00 3:30 PM: Station reports
3:30 3:45 PM: Break
3:45 5:00 PM: Business meeting. All attendees are welcome and invited to attend.
For discussion: Update on BRD Symposium 2014 planning, BRD Symposium fundraising, officer election
5:00 PM: Adjourn

Minutes of the business meeting: Update was given on planning for the BRD Symposium 2014. Symposium will be held on July 30 and 31, 2014, in Denver CO at the Renaissance Denver Hotel, in conjunction with the Summer Academy of Veterinary Consultants meeting. The AVC and AABP have been very supportive; Linda Hofner of Frosch Travel (formerly Cottonwood Travel) in Greely CO, which plans AVC meetings, is assisting with meeting planning. Speakers have all confirmed; list of speakers and agenda can be found at the website: www.brdsymposium.org. Work to secure sponsorship has begun; companies who have not yet been contacted but who might be willing to sponsor were discussed, and various members offered to contact different companies. Chris Chase is chair of Symposium and Amelia Woolums is secretary, with help of Amy Young (UC Davis) and Alison Van Eenennaam they sending letters and invoices to sponsors. Amelia and Amy will keep track of potential sponsors contacted and outcome of contact.

NC1192 committee meeting for 2014 will be held in conjunction with the BRD Symposium; plan will be to have a one-half to one day meeting after the Symposium to allow discussion of how the Symposium went, and brief period of discussion of other committee business.

Discussion re NC1192 officers: Chris Chase has been committee chair and Amelia Woolums has been secretary for a few years in a row. This has been mostly because committee membership was quite small for a few years and others participating had too many other commitments to serve. However, committee membership has increased and so now it seems the committee can go back the system used in the past, wherein a new secretary is elected at the annual meeting who will begin serving after the annual report is submitted, and the previous secretary rotates into the chair position after submitting the annual report. The chair will continue to be responsible for organizing the annual meeting, and the secretary will be responsible for collecting and collating the annual reports from each station, and for submitting the annual report on NIMSS. For 2014, Brian Vander Ley, University of Missouri, will be the new secretary of NC1192 and Amelia Woolums will become the chair.

Minutes submitted by Amelia Woolums

Accomplishments

CA obtained funding to perform a Risk assessment, welfare analysis, and extension education for dairy calf respiratory disease management in California. CA also completed studies demonstrating the role of prostaglandins in initiation and progression of lung inflammation in bovine respiratory disease caused by BRSV and H. sonmi. They showed that the inflammation was due at least in part by BRSV; and support the rationale for the use of non-steroidal anti-inflammatory drug use in treatment. CA completed work using microarray analysis of in vitro infection of bovine alveolar epithelial cells with BRSV followed by treatment with concentrated culture supernatant (CCS) from H. somni, revealing which genes were synergistically up regulated. As part of the Coordinated Agricultural Project (CAP) awarded to Texas A & M (PI James Womack), CA performed individual pathogen infection studies to provide mRNA for use in determination of gene usage in host response to each pathogen. CA worked with WA to develop a modular curriculum on BRD and cow-calf operations. See http://extension.wsu.edu/vetextension/brd/Pages/default.aspx CA, GA, and SD submitted a conference grant proposal to USDA NIFA, which was awarded $10,000 to support the 2014 BRD symposium. GA, KS, NE, FL, and MS, completed a survey of cow-calf producers to determine risk factors for preweaning BRD in beef calves. The same group also conducted a survey of veterinarians in cow-calf practice regarding their knowledge and experience with preweaning BRD. The results were presented as part of an afternoon session at the 2013 AABP Convention, with a panel discussion generating good interaction from the audience. KS continued work to develop diagnostic tools for the recognition of multiple viral and bacterial pathogens involved in BRD, and to identify changes in behavior or biomarkers that could lead to objective assessment tools for management of cattle with BRD during extreme summer conditions. Between 2009 to 2011 KS identified a trend towards increasing multi-drug resistance in Mannheimia haemolytica isolates obtained from the Kansas Veterinary Diagnostic Laboratory. LA incorporated BRD research findings into extension programming for beef and dairy producers including the Louisiana Beef Quality Assurance Certification program and the LSU AgCenter Master Cattle Producer Program. MI completed work indicating that testing of blood collected from cattle at slaughter by the bovine TB gamma interferon ELISA was feasible and provided accurate information regarding the TB exposure status of cattle at the plant, a major point of cattle concentration. This has the possibility of improving control and eradication of bovine TB. MI completed their work with the regional BVDV eradication program in the Upper Peninsula. The resuts of the program, which increased testing and identification of BVDV PI animals in the UP, was published in JAVMA. NE in collaboration with LA completed research indicating that the BHV1 ORF2 product promotes BHV1 latency by promoting survival of infected neurons and through DNA binding, which increases the half life of the ORF2 product. Work was also completed showing that the viral VIP 16 and bICP0 are activated in a dexamethasone model of stress-induced latency reactivation. NE and MS are working together to test cattle in BRD outbreaks for novel respiratory viruses that may contribute to BRD but are at this time not recognized. OK completed research using whole genome sequencing to identify single nucleotide polymorphisms (SNPs) that permitted the distinguishing of 8 MLV BHV-1 strains and 14 field BHV-1 strains. Using the SNPs pattern there were grouping for the MLV vaccines. Some of the field strains were identical to selected MLV strains. OK and NADC identified bovine coronavirus (BoCV) from outbreaks of BRD, and sequenced a region of the envelope Spike protein genome sequenced. This showed that BoCV from respiratory disease outbreaks were from a newly identified clade, clade 2; this is in contrast to BoCV in the licensed BoCV MLV vaccine and the historical enteric isolates, which were BoCV clade 1. MARC continued studies to characterize genetic influences on passive transfer of antibody from cow to calf. During the spring and summer of 2013, failure of passive transport of immunoglobulin from cow to calf case and matched control DNAs were scanned to identify areas of the genome associated with failure. MARC completed the first complete genetic sequence of a fully circularized genome of M. haemolytica, and continued characterization of a robust, rapidly-growing bovine monocyte-derived cell line. SD completed research showing that cytopathic (CP) and noncytopathic (NCP) BVDV induce autophagy in infected cells, and that suppression of autophagy decreased viral replication. Work also showed that BVDV strains vary in their effect on NK cell activation, with more virulent BVDV strains inducing different profiles of cell surface marker expression than less virulent strains. SD with collaborators at TX showed that CP and NCP BVDV strains vary in their effects on dendritic cell expression of activation markers, with NCP strains decreasing expression of activation markers, and CP strains increasing expression. VA continued work to develop diagnostic assays that are needed for rapid and accurate identification of H. somni in clinical samples and following laboratory isolation. Optical fiber biosensors are being utilized to develop culture-free, rapid diagnostic tests, VA continued work to characterize the P. multocida exopolysaccharide (EPS) and the biofilm matrix (EPS and proteins) of the biofilms of H. somni and P. multocida grown together in a bioreactor. WA with collaborators has continued to collect samples from beef and dairy cattle that have BRDC and healthy controls for genotyping to identify genomic regions associated with BRD susceptibility. To date, over 3800 cattle have been sampled, with genotyping of 777,000 single nucleotide polymorphisms. WI continued research to determine how neutrophil extracellular traps are induced by H. somni and M. hemolytica. WI also showed that BHV-1 infection of respiratory epithelial cells led to release of factors that impaired formation of extracellular traps in some but not all circumstances, and that macrophages can remodel extracellular traps by use of DNAse II. WI continued research to determine how bovine respiratory epithelial cells respond to infection with BHV-1, M. haemolytica, or both pathogens together, and showed that cells produced a different pattern of inflammatory mediators when they were infected with both pathogens, as compared to either pathogen alone. Funding leveraged from project activities: New Approaches to Bovine Respiratory Disease Prevention, Management, and Diagnosis Conference Grant. USDA National Institute of Food and Agriculture Competitive Grant no. 2013- 01236. 9/30/201312/31/2014. $10,000. A. L. Van Eenennaam (PD). Risk Assessment, welfare analysis, and extension education for dairy calf respiratory disease management in California. UC ANR Strategic Initiative Competitive Grant Program. $599,872. 9/1/2012-8/31/2016 Sharif Aly, California (PD). A metagenomic analysis of the effect of transportation stress and pathogen infection on the nasal bacterial microbiota of cattle. UC Davis Academic Federation Innovative Development Award Program. $7,908 7/1/2012-6/30/2014. A. L. Van Eenennaam (PD). Characterizing the nasal microflora of diseased and healthy cattle. UC Davis Genome Center Core Facility Pilot Projects. $2,000 10/1/2012-10/1/2014. A. L. Van Eenennaam (PD) Integrated program for reducing bovine respiratory disease complex in beef and dairy cattle. USDA National Institute of Food and Agriculture Competitive Grant no. 2011-68004-30367. 4/15/20114/15/2016. $9,750,000. Jim Womack, Texas (PD). A case-control study to determine herd-level risk factors for nursing calf bovine respiratory disease (BRD) on cow-calf operations. Woolums A, Smith DR, Berghaus R, Daly R, White B, Stokka J. Zoetis. November 2012-December 2014. $42,960 (amount corrected from 2012 GA station report). USDA, NIFA, Analysis of Bovine Herpesvirus 1 stress induced reactivation from latency. 10/1/2013-9/30/207. The goals of this grant are to identify viral and cellular genes that are stimulated during the early stages of stress-induced reactivation from latency. C. Jones. USDA, NIFA, Analysis of viral factors that regulate the bovine herpesvirus 1 latency-reactivation cycle. 10/01/09-9/30/13. The goals of this grant are to test whether a protein encoded by the BHV-1 LR (latency related) gene controls the latency-reactivation cycle. Additional studies will identify and characterize a micro-RNA encoded within the LR gene. C. Jones. Nebraska Research Initiative, Dynamics of acquisition and transmission of poly-microbial respiratory disease that affect cattle: bovine respiratory disease complex (BRDC). 7/1/2012-6/30/2014. The goals of this grant are to compare viruses in cattle that are suffering from BRDC versus healthy cattle. C. Jones. Boehringer Ingelheim Vetmedica, Inc. Development of a LR mutant/gE minus BHV-1 modified live vaccine. 6/1/2012-5/30/2014. C. Jones. R. Fulton, Principal Investigator Bovine Herpesvirus-1: Molecular Characterization of Vaccine, Reference, and Field Strains. Novartis Animal Health Greensboro, NC.$165,430. - Current. R. Fulton, Principal Investigator. Bovine Coronavirus Vaccine Strains: Diversity of Field Strains for Vaccine and Diagnsostic Test Development. Oklahoma State University Technology Business Development Program. $20.000. 2011-2013. R. Fulton, Principal Investigator. Bovine Coronavirus Respiratory Challenge in Neonate Calves. $143,112.50, Pfizer Animal Health, Kalamazoo, MI. 2012-Current. R. Fulton, Principal Investigator. Bovine Herpesvirus-1: Evaluation of Genetic Diversity of Field Strains From Various Clinical Forms. Novartis Animal Health, Greensboro, NC. $36,450. 2013- Current. R. Fulton, Principal Investigator, Bovine Herpesvirus-1: Selection of Genetic Variants for Vaccine Development and Evaluation. Oklahoma State University Technology Business Development Program. $25,000. 2013-2014. T. Confer, Principal Investigator 2009 2013 - Mannheimia haemolytica chimeric protein vaccine for delivery of multiple outer membrane protein and leukotoxin antigens. USDA-CSREES, AFRI Competitive Grant (Grant # 2009-01626) - $375,000 T. Confer, Principal Investigator (J. Taylor, Co-PI) 2010 - Comparison of Mannheimia haemolytica isolates from cattle in Australia with U.S. isolates. Pfizer Animal Health, Kalamazoo, MI & Australia - $75,419 SBIR Phase I: High Sensitivity Optical Fiber Biosensor with Nanoscale Coatings for Rapid Diagnostics of MRSA. NSF SBIR. $144,446; subaward to T. Inzana. Polymicrobial Biofilm Formation by Pasteurella multocida and Histophilus somni in vitro and in the Bovine Host. Internal Research Competition, College of Veterinary Medicine, Virginia Tech. $20,000, 7/1/13-6/30/15. Development of a DNA-based nanoscale optical fiber biosensor assay to detect Brucella. T.J. Inzana, R. Heflin, and A. Bandara. United States Department of Agriculture. $200,000. Biofilm formation by Pasteurella multocida and its co-habitive interaction with Histophilus somni biofilm in vitro and in the bovine host. USDA-NIFA, $499,999, 10/1/2013-9/30/17. Improving Fertility of Dairy Cattle using Translational Genomics. Agriculture and Food Research Initiative. US Department of Agriculture. PI: T.E. Spencer and Co-PIs: H.L. Neibergs, J.B. Cole, J. C. Dalton, A.J. De Vries, P.J. Hansen, D.A. Moore

Publications

Agnes JT, Zekarias B, Shao M, Anderson ML, Gershwin LJ, Corbeil LB. Bovine respiratory syncytial virus and Histophilus somni interaction at the alveolar barrier. Infect Immun. 2013 Jul;81(7):2592-7. doi: 10.1128/IAI.00108-13. Van Eenennaam, A. L. 2012. Integrated program for reducing bovine respiratory disease complex in beef and dairy cattle coordinated agricultural project (BRD CAP). The American Association of Bovine Practitioners (AABP) Proceedings 45: 36-39. Van Eenennaam, A. L. 2012. The Potential Value of DNA-based Tests for Bovine Respiratory Disease (BRD) Resistance to the Beef Cattle Supply Chain The American Association of Bovine Practitioners (AABP) Proceedings 45: 60-65. Heins BD, Nydam DV, Woolums AR, Berghaus RD, Overton MW. Comparative efficacy of enrofloxacin and tulathromycin for treatment of pre-weaning respiratory disease in dairy heifers. J Dairy Sci 2013. Accepted. Woolums AR, Berghaus RD, Smith DR, White BJ, Engelken TJ, Irsik MB, Matlick DK, Jones AL, Ellis RW, Smith IJ, Mason GL. A producer survey of herd-level risk factors for nursing beef calf respiratory disease. J Am Vet Med Assoc 2013. 243:538-547. Theurer M.E., Anderson D.E., White B.J., Miesner M.D., Mosier D.A., Coetzee J.F., Lakritz J., Amrine D.E. Effect of Mannheimia haemolytica pneumonia on behavior and physiologic responses of calves experiencing hyperthermal environmental conditions. J Anim Scii 2013.91:1-13. Hanzlicek G.A., Renter D.R., White B.J., Wagner B.A., Dargatz D.A., Sanderson M.W., Scott H.M., Larson R.E.. Management practices associated with the rate of pre-weaning calf respiratory disease: results from a national survey of U.S. cow-calf operations. 2013 J Am Vet Med Assoc. 242(9): 1271-1278. PMID: 23600786 Babcock A.H., White B. J., Renter D. G., Dubnicka S., Scott H.M. Predicting cumulative risk of bovine respiratory disease complex using feedlot arrival data and daily morbidity and mortality counts. Can J Vet Res. 2013. 77(1):33-44. Fraser BC, Anderson DE, White BJ, Miesner MD, Wheeler C, Amrine D, Lakritz J, Overbay T. Assessment of a commercially available point-of-care assay for the measurement of bovine cardiac troponin I concentration. Am J Vet Res. 2013 Jun; 74 (6):870-3 Amrine D.E., White B.J., Larson R.L, Anderson D.E, Mosier D.A, Cernicchiaro N. 2013. Precision and accuracy of clinical illness scores, compared with pulmonary consolidation scores, in Holstein calves with experimentally induced Mycoplasma bovis pneumonia. Am J Vet Res. 74:310-315. PMID: 23363359 Hanzlicek G.A., Lubbers B.V. Antimicrobial multidrug resistance and coresistance patterns of Mannheimia haemolytica isolated from bovine respiratory disease cases a three-year (2009-2011) retrospective analysis. J Vet Diag Invest, 25:413-417, 2013. Cernicchiaro N., White B.J., Renter D.G., Babcock A.H. Evaluation of economic and performance outcomes associated with the number of treatments after an initial diagnosis of bovine respiratory disease in commercial feeder cattle. Am J Vet Res, 74:300-309, 2013. Rainbolt S., Moore M., Lubbers B., Davis R., Pillai D., Mosier D. Comparison of Mannheimia haemolytica isolates from an outbreak of Bovine Respiratory Disease. Merial-NIH Veterinary Scholars Symposium, East Lansing, MI, 2013. Corbett EM, Grooms DL, Bolin SR. Evaluation of Skin Samples by RT-PCR Following Immunization with a Modified-Live Bovine Viral Diarrhea Virus Vaccine. Am J Vet Res. 2012;73(2):319-324. Okafor CC, Grooms DL, Bolin SR, Kaneene JB. Detection of bovine interferon-³ response in blood collected during exsanguination of cattle sensitized with Mycobacterium bovis. Amer J Vet Res 2012:73(6):847-853. Lim A, Steibel JP, Coussens PM, Grooms DL, Bolin SR. Differential Gene Expression Segregates Cattle Confirmed Positive for Bovine Tuberculosis from Antemortem Tuberculosis Test-False Positive Cattle Originating from Herds Free of Bovine Tuberculosis. Vet Medicine International. 2012;2012:192926. Urban-Chmiel R, Grooms DL. Prevention and Control of Bovine Respiratory Disease. Journal of Livestock Science. 2012;3:27-36. Okafor CC, Grooms DL, Bolin SR, Gravelyn TD, Kaneene JB. Factors that can affect measurable interferon-³ response to Mycobacterium bovis in cattle at time of slaughter. J Vet Diagn Invest, 2013 Feb 15. [Epub ahead of print]. Okafor CC, Grooms DL, Bolin SR, Averill JJ, Gravelyn TD, Kaneene JB. Bovine TB INF-³ Assay at Slaughter: A Novel Strategy for Targeted Surveillance. Transboundary and Emerging Diseases, 22 MAR 2013, DOI: 10.1111/tbed.12080. Urban-Chmiel R, Grooms DL .Rapid Detection of Bovine Respiratory Syncytial Virus in Poland Using A Human Patient-Side Diagnostic Assay. Transboundary and Emerging Diseases. 2013 Aug 12. doi:10.1111/tbed.12134. Grooms DL, Barlett BB, Bolin SR, Corbett EM, Grotelueschen DM, Cortese VS. Special Report: A Review of the Michigan Upper Peninsula Bovine Viral Diarrhea Virus Eradication Project. J Am Vet Med Assoc. 2013 Aug 15;243(4):548-54. doi:10.2460/javma.243.4.548. Grooms DL, Brock KV, Bolin SR, Grotelueschen DM, Cortese VS. Effect of constant exposure to cattle persistently infected with bovine viral diarrhea virus on morbidity, mortality, and performance in feedlot cattle: summary of three studies. J Am Vet Med Assoc. Accepted for Publication, 2013. Sinani, D., L. Frizzo da Silva, and C. Jones. 2013. A bovine herpesvirus 1 protein expressed in latently infected neurons (ORF2) promotes neurite sprouting in the presence of activated Notch1 or Notch3. J of Virology, 87:1183-1192. Pittayakhajonwut, D., D. Sinani, and C. Jones. 2013. A protein (ORF2) encoded by the latency related gene of bovine herpesvirus 1 interacts with DNA. J of Virology, 87: 5943-5501. Jones, C. 2013. Bovine herpesvirus 1 (BHV-1) and herpes simplex virus type 1 (HSV-1) promote survival of latently infected sensory neurons, in part by inhibiting apoptosis. J of Cell Death, 6:1-16. Frizzo da Silva, L., I. Kook, A. Doster, and C. Jones. Bovine herpesvirus 1 regulatory proteins, bICP0 and VP16, are readily detected in trigeminal ganglionic neurons expressing the glucocorticoid receptor during the early stages of reactivation from latency. In Press, J of Virology. Confer AW, Ayalew S. The OmpA Family of Proteins: Roles in Bacterial Pathogenesis and Immunity. Vet Microbiol. 163: 207-222, 2013 Taylor JD, Doyle DJ, Confer AW. Use of REP-PCR and 16s rRNA sequencing for comparison of Mannheimia haemolytica isolates obtained from fatal cases of bovine respiratory disease in the United States and Australia. Austral Vet J, 2013, in press Kraus, B., Fulton, R.W., Johnson, B.J., Sjeklocha, D.B.: Case Report- Comparison of Pooled Polymerase Chain Reaction Testing to Non-Pooled Antigen Capture Enzyme-Linked Immunosorbent Assay Using Individual Samples to Detect Bovine Viral Diarrhea Virus Persistently Infected Stocker Calves. Bovine Practitioner, 46: 131-135, 2012. Fulton,R.W.: Host Response to Bovine Viral Diarrhea Virus and Interactions with Infectious Agents in the Feedlot and Breeding Herd. Biologicals, 41:31-38, 2013. Fulton, R.W., Ridpath, J.F., Burge, L.J.: Bovine Coronaviruses From the Respiratory Tract: Antigenic and Genetic Diversity. Vaccine, 31: 886-892,2013. dOffay, J.M., Fulton,R.W., Eberle, R.: Complete Genome Sequence of the NVSL BoHV-1.1 Cooper Reference Strain. Archives of Virology, 158: 1109-1113, 2013. Fulton, R.W., dOffay, J.M., Eberle, R.: Bovine Herpesvirus-1: Comparison and Differentiation of Vaccine and Field Strains Based on Genomic Sequence Variation. Vaccine, 31:1471-1479, 2013 . Chase CCL. Viral Disruption of Adaptive Immunity. 2013. Biologicals 41:52- 60. Perry GA, AD Zimmerman, RF Daly, RE Buterbaugh, J Rhoades, D Scholz, A Harmon, CCL Chase. 2013. The effects of vaccination on serum hormone concentrations and conception rates in synchronized naive beef heifers.Theriogenology 79:200-205. Elswaifi, S.F., W.K. Scarratt, T.J. Inzana. 2012. The role of lipooligosaccharide phosphorylcholine in colonization and pathogenesis of Histophilus somni in cattle. Vet. Res. 43:49. Inzana, T.J., R. Balyan, and M. D. Howard. 2012. Decoration of Lipooligosaccharide with N-acetyl-5-neuraminic acid contributes to Histophilus somni virulence and resistance to host defenses. Vet. Microbiol. 161:113-121. Aulik, N.A., D. Atapattu, D. McCaslin and C. J. Czuprynski. 2013. Brief heat treatment causes a structural change and enhances cytotoxicity of the Escherichia coli ±-hemolysin. Immunopharmacol Immunotoxicol. 35:15-27. Hellenbrand, K.M., K.M. Forsythe, J.J. Rivera-Rivas, C.J. Czuprynski, and N.A. Aulik. 2013. Histophilus somni causes extracellular trap formation by bovine neutrophils and macrophages. Microb Pathog. 54:67-75. N'jai, A.U., J. Rivera, D.N. Atapattu, K. Owusu-Ofori, and C.J. Czuprynski. 2013. Gene expression profiling of bovine bronchial epithelial cells exposed in vitro to bovine herpesvirus 1 and Mannheimia haemolytica. Vet. Immunol. Immunopathol. Jul 1. doi:pii: S0165- 2427(13)00193-1. 10.1016

Impact Statements

A validated calf BRD scoring system will be used to identify husbandry practices associated with BRD. When practices associated with BRD are identified, a tool to can be developed to provide animal health professionals a method to identify and change husbandry procedures to improve animal health.
The ongoing studies on pathogenesis of the synergy between BRSV and H. somni will yield information valuable for further development of therapeutic and prophylactic regimens.
The RNA sequences collected in specific pathogen infections performed for the CAP collaboration will demonstrate which host response genes are activated for each primary infective organism. This information will be used to determine genotypes of cattle that are resistant to BRD.
The study of risk factors for BRD in preweaning beef calves will provide veterinarians and producers with information to allow development of better methods of management to prevent calf BRD.
The 2014 BRD Symposium will provide an opportunity for veterinarians, scientists, policy makers, and producers to learn about the latest scientific findings related to BRD. This should improve field application of new findings and should also provide new opportunities for collaboration among researchers. Together this should help veterinarians and producers institute new practices to decrease the negative impacts of BRD on health and growth of cattle.
Because accurate field diagnosis of BRD can sometimes be difficult, the development of clinical illness scores, behavior monitoring systems, and specific biomarkers to more accurately identify BRD will help improve the ability of producers to identify and treat cattle with BRD more quickly and effectively.
Monitoring the emergence of multi-drug resistance in M. haemolytica isolates will help define the need to modify treatment approaches to preserve antimicrobial utility in the therapy of BRD.
The regional BVDV eradication program led to testing approximately one half of the herds in the Michigan UP, with identification of BVDV PI cattle in 3.9% of herds. As a result of the project, mandatory BVDV testing of cattle participating in the UP State Fair has been initiated, which will decrease the risk of BVDV transmission in cattle involved in the Fair and which will improve the knowledge of cattle producers regarding a practice important to maintenance of cattle health.
Validating the use of the IFN-gamma ELISA to identify cattle infected with TB at slaughter provides an efficient and feasible method to improve identification of TB-infected cattle at points of concentration, which should advance bovine TB control and eradication.
The research identifying viral proteins required for viral latency and latency reactivation will lead to the development of safer and more effective BHV1 vaccines, which should prevent disease by limiting latency.
The research to test cattle in BRD outbreaks for novel viruses will help determine whether new viruses contribute to BRD. If new viruses are found, information regarding these viruses will aid the development of new vaccines to better prevent BRD.
The BoHV-1 sequencing permitted the identification of SNPs which allow for the differentiation of the MLV strains from field strains. This permits more accurate diagnosis of BoHV-1 for the clinician and cattle owner.
BoCV strains isolated from BRD cases manifest genetic and antigen differences. These differences are unique in that the licensed MLV vaccine in the U.S is a different clade. The current enteric strain BoCV1 in the vaccine should be shown to be efficacious for the BoCV2 respiratory strains found in the current study. Or perhaps there should be BoCV2 vaccines made for BRD.
The work to determine whether genetic factors influence transfer of antibody from cow to calf will reveal whether cattle can be selected that pass high concentrations of antibody to their calves, thus helping these calves be more resistant to disease.
The complete sequence of a full circularized M. haemolytica genome will allow determination of genetic factors that help this bacteria cause BRD, which will help scientists develop tests to identify strains of the bacteria more likely to cause disease, and which will also aid in the development of vaccines that better protect cattle from disease due to M. haemolytica.
The work to develop fiber optic biosensors may lead to the development of highly sensitive and mobile detection device that will identify H. somni in clinical samples. This should help veterinarians treat and prevent disease better by allowing them to identify the cause of disease more quickly and inexpensively.
The research on biofilm formation by P multocida and H somni will reveal new methods these bacteria persist in the host, causing disease. Identification of these methods will allow the discovery of new therapeutic and preventative strategies to decrease BRD.
The identification of genetic regions associated with BRD susceptibility provides the first step in selecting for cattle that are less likely to have this disease.
Studies identifying the impact of BVDV infection on autophagy and on expression of activation markers by dendritic cells provides new foundational knowledge regarding a common and important pathogen, and provides the basis for the development of improved means to counteract and prevent immunosuppression due to BVDV.
The information regarding formation of extracellular traps improves foundational knowledge regarding the bovine immune response to infection, and will provide the basis for future studies to improve the ability of cattle to resist respiratory infection.
Results of studies to evaluate the response of respiratory epithelial cells to infection improves foundational knowledge regarding how the bovine lung responds to insult, and will allow researchers to develop ways to decrease lung damage and improve resistance of the lung to infection.

Date of Annual Report: 12/09/2014

Report Information

Annual Meeting Dates: 07/29/2014 - 07/29/2014
Period the Report Covers: 10/01/2013 - 09/01/2014

Participants

Amelia Woolums*, University of Georgia; Chris Chase*, South Dakota State University; Chuck Czuprynski*, University of Wisconsin; Holly Neibergs*, Washington State University; Laurel Gershwin*, University of California, Davis;
Derek Mosier*, Kansas State University; Robert Fulton*, Oklahoma State University; Tom Inzana*, Virginia Tech; John Richeson, West Texas A&M University; Amy Young, University of California, Davis; Christine Navarre*, Louisiana State University; Terry Lehenbauer, University of California, Davis;
Dan Grooms*, Michigan State University; Dale Grotelueschen, University of Nebraska, Great Plains Veterinary Education Center; BJ Newcomer, Auburn University; Brian Vander Ley*, University of Missouri.

Brief Summary of Minutes

Meeting date and time: Tuesday, July 29, 2014; 8:30AM-5:00PM
Location: Vail Room of the Denver Renaissance Hotel, Denver, CO

Meeting Agenda:

8:00AM: Registration

8:30AM: Welcome and BRD Symposium Questions/Comments

8:50AM CAP BRD Research Update

9:20AM: Station Reports

10:00AM: Break

10:30AM: Station Reports

12:00PM: Lunch

1:00PM Station Reports

3:15PM Break

3:30PM Business Meeting

5:00PM Adjourn

Minutes of the Meeting:

Meeting Minutes

I. Introduction

a. Attendees were welcomed and reminded of registration fee

b. Attendees introduced themselves

II. Update on BRD Symposium

a. Meeting organized by NC 1192 members and the BRD Cap. The symposium has
been sponsored by AVC and AABP as well as other sponsors. Website introduced as www.brdsymposium.com.

III. BRD Cap Update

a. Update on BRD Cap progress and results presented by Alison Van Enennaam.

IV. Station Reports

a. Dr. B.J. Newcomer presented a report on BRD related research activities
at Auburn University.

b. Dr. Laurel Gershwin presented a report on BRD research at the University of California-Davis.

c. Dr. Amelia Woolums presented a report on BRD research at the University of Georgia.

d. Dr. Derek Mosier presented a report on BRD research at Kansas State University.

e. Dr. Dan Grooms presented a report on BRD research at Michigan State University.

f. Dr. Dave Smith presented a report on BRD research at Mississippi State University.

g. Dr. Brian Vander Ley presented a report on BRD research at the University of Missouri.

h. Dr. Robert Fulton presented a report on BRD research at Oklahoma State University.

i. Dr. Chris Chase presented a report on BRD research at South Dakota State University.

j. A question was presented by Dr. Neibergs regarding the length of time BRD vaccine strain viruses could still be found in clinical samples. Dr. Fulton stated that BHV-1 and BVDV can be isolated from vaccinates for a period of time; however, establishing a definite period of time in which samples would be questionable.

k. Dr. John Richeson presented a report on BRD research at West Texas A&M.

l. Dr. Tom Inzana presented a report on BRD research at Virginia Tech.

m. Dr. Holly Neibergs presented a report on BRD research at Washington State University.

n. Dr. Chuck Czuprynski presented a report on BRD research at the University of Wisconsin.

V. Business Meeting.

a. Dr. Woolums introduced Dr. Neal Merchen as the new administrative advisor.

b. Peter Johnson sends his regrets.

c. New BRD Project Proposal

d. The members of the project decided to select a 3-4 person committee to write the new BRD project proposal. Chris Chase, Dan Grooms, and Brian Vander Ley will serve as the committee to rewrite the proposal.

e. Dr. Woolums presented information from Chris Hamilton that may be useful in formulating the new project.

f. Update presented on station reports and the suggested template from NCRA. Stations will be sent the updated template next year’s reports to facilitate inclusion of the information requested in the template.

g. Nominations for a new secretary were requested. Dan Grooms offered to serve as the next secretary for NC 1192.

h. Next year’s NC 1192 has been tentatively scheduled for September 14 and 15, 2015.

i. Members discussed other institutions that should be invited to participate in NC 1192. Members are encouraged to reach out to BRD researchers who are not currently participating in NC 1192.

j. Meeting was adjourned.

Minutes submitted by Brian Vander Ley

Accomplishments

CA completed and published an enhanced BRD scoring system based on dichotomous variables that is designed to improve feasibility for on-farm use. Committee participants from CA, GA, LA, NE, OK, SD, and USDA MARC along with representatives from the National Cattlemen’s Beef Association and the Texas Veterinary Diagnostic Lab to organize and execute the 2014 BRD Symposium in conjunction with the summer Academy of Veterinary Consultants meeting in Denver, CO (July 30-31). GA has demonstrated increased IL-17 production and decreased IL-10 production following exposure of PBMCs to clinically virulent BVDV strains compared to less virulent BVDV strains. Differential responses in lymphocyte gene expression following exposure to high and low virulence BVDV strains were also observed. GA has demonstrated association of BRSV N protein with host cell MDA-F. Work to elucidate the impact of this association on IFN production is on-going. KS demonstrated decreased pulmonary damage and fewer clinical signs illness in heifers treated with tildipirosin compared to tulathromycin when both were used as metaphylactic agents. KS developed a multiplex PCR assay capable of detecting BVDV, BRSV, BCoV, BHV-1, M. haemolytica, P. multocida, H. somni, B. trehalosi, and Mycoplasma bovis. KS investigated transmission of M. haemolytica between 40 auction-derived calves and found that calves initially found to be culture negative would shed two distinct strains despite being challenged with a single strain after initial culture. MO has demonstrated an association between clinically diagnosed field cases of BRD and increases in lipopolysaccharide binding protein and haptoglobin in feedlot cattle. Transferrin concentrations were not significantly associated with BRD status. MS found that bull calves and calves born to young dams were more likely to develop BRD prior to weaning. MS used a proteomics approach to identify interaction of 97 bovine proteins with the NS3 protein from CP BVDV. These interactions are thought to manipulate cellular translation proteins and may be useful targets for future antiviral therapies. MS demonstrated that metaphylaxis decreased BRD incidence and increase dietary protein resulted in higher average daily gain in high risk stocker calves. NE demonstrated that in BHV-1 infections, ORF2 proteins fuse with reading frame B (15d ORF) and was more stable in transfected cells. This binding stimulated neurite formation in mouse neuroblastoma cells and interfered with Notch3 mediated trans-activation. Increased stability of 15d ORF is predicted to enhance the establishment of latency. NE found two regulatory viral proteins (VP16 and blCP0) that are expressed within 90 minutes of dexamethasone treatment of calves latently infected with BHV-1. NE demonstrated that BHV-1 infection stimulated inflammasome formation which is predicted to impact clinical symptoms in cattle. OK isolated vaccine strain BVDV and BHV-1 from nasal secretions and lung samples from modified live virus vaccinated calves. OK provided further evidence that respiratory coronavirus infections of cattle are caused by coronaviruses from either BoCV2 or BoCV3 clades. OK demonstrated that M. haemolytica produces an IgA protease that is immunogenic in cattle. SD characterized the development and dissemination of a cytopathic BVDV under natural conditions. SD demonstrated that BVDV replication did not occur in autophagosomes but that the autophagy inhibiting drug, 3MA, suppressed viral production. SD demonstrated that bovine NK cells are susceptible to BVDV infection but not to the production of infectious virus. BVDV infection results in NK phenotypic and activation changes that are strain dependent and may result in immunosuppression. SD discovered a novel protein interaction between BVDV Npro and cellular S100 A9 protein that appears to inhibit type I interferon production. SD attempted to infect bovine monocyte derived dendritic cells with BVDV and found that fully mature DCs fail to produce infection BVDV following infection. SD demonstrated that the infection of calves with bovine virus diarrhea virus (BVDV) is a transient and self-limiting infection that can result in a period of humoral immunosuppression with the type of TH response affected dependent up on the viral strains. VA developed an ELISA to detect bovine antibodies to H. somni exopolysaccharide. VA demonstrated that capsule formation in P.multocida interferes with biofilm formation. VA reported the putative genes believed to be responsible for P.multocida exopolysaccharide production. WI demonstrated that M. haemolytica produces greater amounts of biofilm when incubated in RPMI-1640 media and that biofilm formation could be inhibited by addition of anti-OmpA antibodies, galactose, or mannose to the growth medium. WI found that neutrophils attach to and form NETs at the edge of biofilm aggregates rather than on the apical surface of the biofilm mass. WI demonstrated that bovine macrophages use DNase II to degrade extracellular traps. WI discovered a spontaneously transformed bovine macrophage cell line that has been characterized as CD18+, phagocytic, esterase+, and cytokeratin-. Funding leveraged from project activities Comparison of nasopharyngeal swabs, pharyngeal recess swabs, bronchoalveolar lavage, and transtracheal wash for detecting bacterial and viral pathogens in dairy calves with bovine respiratory disease. Lehenbauer TW, Doyle DJ, Woolums AR, Aly SS, Champagne JD, Blanchard PC, Crossley BM. UC Davis Center for Food Animal Health. February 2014 – January 2015. Awarded $20,000. Effect of injectable trace minerals on the humoral and cell-mediated immune responses to vaccine antigens following administration of a modified-live viral vaccine in dairy calves. Palomares RA, Hurley DJ, Woolums AR. Multimin-USA. January 2014 – December 2014. Awarded $56,967. Webinars in bovine immunity. Woolums AR. Veterinary Technical Services, Bayer Animal Health GmbH. January – December 2014. Awarded $10,200. Webinars in bovine innate immunity. Woolums AR. Veterinary Technical Services, Bayer Animal Health. January – December 2013. Awarded $8,000. A case-control study to determine herd-level risk factors for nursing calf bovine respiratory disease (BRD) on cow-calf operations. Woolums A, Smith DR, Berghaus R, Daly R, White B, Stokka J. Zoetis. November 2012-December 2014. $42,960 Analysis of Bovine Herpesvirus 1 stress induced reactivation from latency. Jones, C. USDA, NIFA, 10/1/2013-9/30/2017. Dynamics of acquisition and transmission of polymicrobial respiratory disease that affects cattle: bovine respiratory disease complex (BRDC). Jones, C. Nebraska Research Initiative, 7/1/2012-6/30/2014. Development of a LR mutant/gE minus BHV-1 modified live vaccine. Jones, C. Boehringer Ingelheim Vetmedica, Inc., 6/1/2012-5/30/2014. R.W. Fulton. Principal Investigator “Bovine Herpesvirus-1: Molecular Characterization of Vaccine, Reference, and Field Strains”. Novartis Animal Health Greensboro, NC.$165,430. - Current. R.W. Fulton. Principal Investigator. “ Bovine Coronavirus Vaccine Strains: Diversity of Field Strains for Vaccine and Diagnostic Test Development”. Oklahoma State University Technology Business Development Program. $20.000. 2011-2013. R.W. Fulton. Principal Investigator. “Bovine Coronavirus Respiratory Challenge in Neonate Calves”. $143,112.50, Pfizer Animal Health, Kalamazoo, MI. 2012-Current. R.W. Fulton. Principal Investigator. “Bovine Herpesvirus-1: Evaluation of Genetic Diversity of Field Strains From Various Clinical Forms”. Novartis Animal Health, Greensboro, NC. $36,450. 2013- Current. R.W. Fulton. Principal Investigator, “Bovine Herpesvirus-1: Selection of Genetic Variants for Vaccine Development and Evaluation”. Oklahoma State University Technology Business Development Program. $25,000. 2013-2014. A.W. Confer. Principal Investigator – 2009 – 2013 - Mannheimia haemolytica chimeric protein vaccine for delivery of multiple outer membrane protein and leukotoxin antigens. USDA-CSREES, AFRI Competitive Grant (Grant # 2009-01626) - $375,000 A.W. Confer. Co-Principal Investigator (J. Taylor, Co-PI) – 2010 - Comparison of Mannheimia haemolytica isolates from cattle in Australia with U.S. isolates. Pfizer Animal Health, Kalamazoo, MI & Australia - $75,419 AW. Confer, Principal Investigator – 2013-2014 - Development of a Mannheimia haemolytica model for studying RecA inhibitors. Noble Foundation, Ardmore, OK - $88,384 SBIR Phase I: High Sensitivity Optical Fiber Biosensor with Nanoscale Coatings for Rapid Diagnostics of MRSA. NSF SBIR. $144,446; subaward to T. Inzana. A stocker cattle receiving system to test interactions of health and nutrition for Mississippi stocker calves: Phase I. Effect of crude protein levels and metaphylaxis on growth and performance of newly received stocker calves. MSU Special Research Initiative. Karisch B, Smith DR, Huston C, Vann R. 12/01/12-6/30/14. $50,000. A case-control study to determine herd-level risk factors for nursing calf bovine respiratory disease (BRD) on cow-calf operations. Woolums A, Smith DR, Berghaus R, Daly R, White B, Stokka J. Zoetis. 11/01/12-12/01/15. $42,960.

Publications

Banse H, Woolums AR, Step DL. A review of host pulmonary defenses with reference to cattle. 2014. Bov Pract 48:13-24. Woolums AR, Berghaus RD, Smith DR, White BJ, Engelken TJ, Irsik MB, Matlick DK, Jones AL, Smith IJ. A survey of veterinarians in 6 U.S. states regarding their experience with nursing beef calf respiratory disease. 2014. Bov Pract 48:26-36. Idoate, I. “Acute phase proteins in naturally occurring respiratory disease of feedlot cattle.” Master of Science Thesis. In Press. Sinani, D., L. Frizzo da Silva, and C. Jones. 2013. A bovine herpesvirus 1 protein expressed in latently infected neurons (ORF2) promotes neurite sprouting in the presence of activated Notch1 or Notch3. J of Virology, 87:1183-1192. Pittayakhajonwut, D., D. Sinani, and C. Jones. 2013. A protein (ORF2) encoded by the latency related gene of bovine herpesvirus 1 interacts with DNA. J of Virology, 87: 5943-5501. Frizzo da Silva, L. I. Kook, A. Doster, and C. Jones. 2013. Bovine herpesvirus 1 regulatory proteins, bICP0 and VP16, are readily detected in trigeminal ganglionic neurons expressing the glucocorticoid receptor during the early stages of reactivation from latency. J of Virology, 87: 11214-11222. Wang, J., Alexander, M. Wiebe, and C. Jones. 2014. Bovine herpesvirus 1 productive infection stimulates inflammasome formation and caspase 1 activity. Virus Research, 185: 72-76. Sinani, D., Y. Liu, and C. Jones. 2014. Analysis of a bovine herpesvirus 1 protein encoded by an alternatively spliced latency related (LR) RNA that is abundantly expressed in latently infected neurons. Virology, IN PRESS. Fulton, RW. Large Animal Internal Medicine: Fifth Edition, B.P. Smith, Editor, Dr. V. Cortese, Section Editor, Vaccines. “Bovine herpesvirus-1 Vaccines for Cattle in Bovine Respiratory Disease Vaccines Chapter”, pp. 1471-1475. 2014. Taylor, JD, Confer AW. Large Animal Internal Medicine: Fifth Edition, B.P. Smith, Editor, Dr. V. Cortese, Section Editor, Vaccines. “Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Bibersteinia trehalosi.”. pp 1477 – 1481, 2014 Ayalew S, Confer AW, Shrestha B, Wilson AE, Montelongo M. Proteomic Analysis and Immunogenicity of Mannheimia haemolytica Vesicles. Clin & Vaccine Immunol 20:191-196, 2013. Confer AW, Ayalew S. The OmpA Family of Proteins: Roles in Bacterial Pathogenesis and Immunity. Vet Microbiol. 163: 207-222, 2013 Taylor JD, Doyle DJ, Blackall PJ, Confer AW. Use of REP-PCR and 16s rRNA gene sequencing for comparison of Mannheimia haemolytica isolates obtained from fatal cases of bovine respiratory disease in the USA and Australia. Austral Vet J 92:15-23, 2014. Zimmerman, AD, AL Klein, RE Buterbaugh, B Hartman, CL Rinehart, CCL Chase. 2013. Vaccination with a multivalent modified-live virus vaccine administered one year prior to challenge with bovine viral diarrhea virus type 1b and 2a in pregnant heifers. The Bovine Practitioner 47(1):22-33 Hashish, EA, C Zhan, X Ruan, DE Knudsen, CC Chase, RE Isaacson, G Zhou, W Zhang. 2013. A Multiepitope Fusion Antigen Elicits Neutralizing Antibodies against Enterotoxigenic Escherichia coli and Homologous Bovine Viral Diarrhea Virus In Vitro. Clinical and Vaccine Immunology 20(7):1076–1083 Zimmerman, AD, AL Klein, RE Buterbaugh, CL Rinehart, CCL Chase. 2013. Protection against bovine herpesvirus type 1 (BHV-1) abortion following challenge 8 months or approximately1 year after vaccination. The Bovine Practitioner 47(2):73–81. Rajput, M. K., Darweesh, M. F., Park, K., Braun, L. J., Mwangi, W., Young, A. J., & Chase, C. C. 2014. The effect of bovine viral diarrhea virus (BVDV) strains on bovine monocyte-derived dendritic cells (Mo-DC) phenotype and capacity to produce BVDV. Virology Journal, 11(1), 44. doi:10.1186/1743-422X-11- 44. Chase C. Strategic vaccination: Trying to work around the ups and downs of the immune system. Proceedings of 22nd annual Northeast Dairy Production Medicine Symposium, Syracuse NY, March 22-24, 2013, CD-ROM. Chase C. Effective vaccination of the bovine neonate: challenges and opportunities. Interpretive Summary. 25th ADSA Discover Conference on Food Animal Agriculture-New Developments in Immunity, Nutrition, and Management of the Preruminant Calf, Itasca IL, May 28-30, 2013. Pan, Y., T. Fisher, C. Olk, L.B. Corbeil, T.J. Inzana. 2014. Detection of antibodies to the biofilm exopolysaccharide of Histophilus somni following infection in cattle by enzyme-linked immunosorbent assay. Clin. Vac. Immunol. In Press. Boukahil, I., and C.J. Czuprynski. In revision. Characterization of biofilm formation by Mannheimia haemolytica in vitro. A.H. Babcock, N. Cernicchiaro, B.J. White, S.R. Dubnicka, D.U. Thomson, S.E. Ives, H.M. Scott, G.A. Milliken, D.G. Renter. A multivariable assessment quantifying effects of cohort level factors associated with combined mortality and culling risk in U.S. commercial feedlot cattle. Prev Vet Med. 2013. 108:38-46. PMID: 22871305. M.E. Theurer, R.L. Larson, B.J. White. A meta-analysis of vaccine effectiveness against bovine herpes virus, bovine viral diarrhea virus, bovine respiratory syncytial virus, and parainfluenza-type 3 virus in cattle for bovine respiratory disease complex. J Am Vet Med Assoc (In Print) D.E. Amrine, B.J. White, R.L. Larson, D.A. Mosier. Pulmonary lesions and clinical disease response to Mannheimia haemolytica challenge 10 days following administration of tildipirosin or tulathromycin. J Anim Sci 92:311-319, 2014. D. E. Amrine, B.J. White, R.L. Larson. Comparison of classification algorithms to predict outcomes of feedlot cattle identified and treated for Bovine Respiratory Disease. Comput Electron Agri. 2014 July 105:9-19. Doi: 10.1016/j.compag.2014.04.009. B. Fraser, D.E. Anderson, B.J. White, M.D. Miesner, D.E. Amrine. Associations of various physical and blood analysis variables with experimentally induced Mycoplasma bovis pneumonia in calves. Am J Vet Res. 2014. 75(2): 200-207. doi: 10.2460/ajvr.75.2.200. D.E. Amrine, B.J. White, R. L. Larson, D.A. Mosier. Determining differences in pulmonary lesions and clinical disease response to Mannheimia haemolytica challenge occurring 10 days after administration of tildipirosin, tulathromycin, or saline. 2014 J Anim Sci 92:311-319.

Impact Statements

CA: The work done on BRSV-H. somni synergy by Gershwin and Corbeil has had a significant impact on understanding the effect of infection with BRSV and/or treatment with virulence factors of H. somni on respiratory epithelium. Our previous paper (reported in 2013) demonstrated the role of the matrix metalloproteases on altering permeability at the alveolar interface. Our current studies that derive from the microarray analysis may demonstrate a protective mechanism against viral infection. However, this work is still very preliminary and we are not ready to report fully on the data we have thus far.
GA: If the device for field sampling of respiratory pathogen circulation in nursing beef calves can be validated, it will provide a new method to identify the respiratory viruses and bacteria that contribute to respiratory disease in a population of cattle that are otherwise difficult to sample by traditional methods. A simple and effective method of identification of respiratory pathogens will facilitate research to determine what practices can decrease pathogen circulation in nursing beef calves. The method will also help veterinarians and cattle producers determine what vaccines should be used to control disease in populations of beef calves.
GA: The research to determine how virulence of BVDV strains is related to activation of different T helper cell types will help researchers understand why some strains of BVDV make cattle more sick than other strains. If the pathways that are related to more severe disease can be determined, it may then be possible to develop ways to counteract the effects of the more virulent strains, decreasing disease in infected cattle.
GA: The research to determine how BRSV impairs the host interferon response will provide new foundational knowledge regarding the means by which this virus evades the host immune response. Also, because interferon is a key component in the early stages of the development of an effective anti-viral immune response, it may be that manipulation of BRSV to block the ability of the virus to impair interferon production could lead to the development of vaccines that induce better long-term immunity. This could lead to the development of more effective vaccines to protect cattle from BRSV infection.
GA: Because there is very little original research on risk factors for nursing calf respiratory disease in cow-calf herds, it is difficult for veterinarians to make evidence-based recommendations to producers regarding methods to decrease occurrence of the disease in herds where it is a problem. The study of risk factors for nursing calf respiratory disease has the potential to provide veterinarians and producers with the information they need to develop better methods of management to prevent calf respiratory disease. Decreased rates of respiratory disease will improve calf health and well-being, will decrease the need for antimicrobial use, and should enhance producer profitability.
GA: The 2014 BRD Symposium will provide an opportunity for veterinarians, scientists, policy makers, and producers to learn about the latest scientific findings related to BRD. This should improve field application of new findings and should also provide new opportunities for collaboration among researchers. Together this should help veterinarians and producers institute new practices to decrease the negative impacts of BRD on health and growth of cattle.
LA: Incorporating the latest research data into extension programs increases awareness of the disease and allows beef and dairy producers to incorporate new prevention and treatment modalities into their herd health plans. This in turn reduces the negative effects of this disease on health and welfare of cattle and increases the economic sustainability of individual producers as well as the beef and dairy industries.
MO: The results of our research support previous findings that haptoglobin is significantly associated with both naturally occurring and induced BRD. Additionally, we have now demonstrated that lipopolysaccharide binding protein is significantly elevated in naturally occurring BRD cases. This information provides an improved understanding of the immune response to BRD and provides opportunities for the development of diagnostics to improve BRD diagnosis.
MS: The finding of gender effects on BRD risk in the field is important because it aids in directing basic immunologic research questions. Also, the finding is of practical importance because the information might be used to manage BRD risk in production settings (e.g. by managing heifer calf pairs separately).
OK: Field studies will permit evaluation of current viral and bacterial vaccines along with newly developed vaccines. The identification, cloning, and production of subunit components of M. haemolytica and P. multocida offer opportunity for new bacterial vaccines to control BRD. The diversity (antigenic) of BVDV will be further examined to determine appropriateness/relevance of current and future BVDV vaccines to control BVDV. Bovine coronavirus identified in cattle with BRD represents another infectious agent which either singly or in combination with other viruses and bacteria may have a role in BRD.
OK: The BoHV-1 sequencing permitted the identification of SNPs which allow for the differentiation of the MLV strains from field strains. This permits more accurate diagnosis of BoHV-1 for the clinician and cattle owner. BoCV strains isolated from BRD cases manifest genetic and antigen differences. These differences are unique in that the licensed MLV vaccine in the U.S is a different clade. The current enteric strain BoCV1 in the vaccine should be shown to be efficacious for the BoCV2 respiratory strains found in the current study. Or perhaps there should be BoCV2 vaccines made for BRD. It is important that PI cattle be accurately diagnoses for biosecurity purposes, and tests must be validated for the accuracy. In the current study, the antigen capture ELISA test performed better than the pooled PCR test.
OK: The IgA proteases of M. haemolytica are potential virulence factors for M. haemolytica. Their roles as virulence factors help better explain the virulence of this bacteria in BRD. Potentially these proteins might be used for vaccines protecting cattle against M.haemolytica disease.
VA: Rapid and culture-free diagnostic assays are required for the detection of H. somni in clinical samples. The NOFS assay is capable of detecting about 100 cells of H. somni, and we expect that sensitivity to carry over to clinical specimens. Performance of the assay does not require skill or sophisticated laboratory conditions, and can be completed in less than an hour in a basic laboratory or even under field conditions. Detection of antibodies to the H. somni EPS may be a valuable indicator of active disease due to H. somni, and should be pursued as a supplementary test.
VA: Our results support that H. somni and P. multocida are capable of forming a polymicrobial biofilm, indicating that prevention and treatment options may need to be directed at the biofilm and not planktonic bacteria. Furthermore, there is a clear inverse correlation between capsule production and EPS/biofilm production by P. multocida strains, including those from BRD. We postulate that isolates that are the most encapsulated cause the most serious forms of BRD, and do not form good biofilms. However, these isolates can take advantage of the biofilm formed by H. somni.

Date of Annual Report: 01/29/2016

Report Information

Annual Meeting Dates: 09/16/2015 - 09/16/2015
Period the Report Covers: 10/01/2014 - 09/30/2015

Participants

Amelia Woolums-Mississippi State University
Chris Chase-South Dakota State University
Terry Lehenbauer-University of California-Davis
Roberto Palomares-University of Georgia
Tom Inzana-Virginia Tech
Danielle Doyle-Mississippi State University
Dave Smith-Mississippi State University
Neal Merchen-University of Illinois
Brian Vander Ley-University of Missouri

Brief Summary of Minutes

Meeting Minutes

Room 214 New Orleans Convention Center, New Orleans, Louisiana

September 16, 2015

Attendees:

Amelia Woolums

Chris Chase

Terry Lehenbauer

Roberto Palomares

Tom Inzana

Danielle Doyle

Dave Smith

Neal Merchen

Brian Vander Ley

8:00-8:15: Attendees were welcomed to the meeting, introductions were made, and Dr. Neal Merchen, our administrative assistant introduced himself to the group.

8:15-10:00, 10:15-12:00: The NC 1192 project is set to expire next September. To continue the project, a renewal application will have to be filed. Dr. Vander Ley submitted the request to renew the project along with the issues and justification for the project in the NIMMs system by the September 15, 2015 deadline. The committee discussed the following timeline:

September 15: Request to renew along with Issues and Justification due.

October 15: Project Objectives due.

November 1: Appendix E information completed and submitted by participating stations

December 1: Completed project due to be uploaded in NIMMs.

Following a review of the timeline, committee members elected to request a change in number to NC 107 if possible. This is the original number assigned to the project. Dr. Vander Ley will contact Chris Hamilton to see if the change can be made.

Committee members discussed possibilities for recruiting new members to the committee. Several ideas including reaching outside the veterinary community, holding committee meetings in more convenient places for animal scientists/other disciplines to meet, and using current members’ networks to find new, interested committee members were discussed.

The objectives from the previous project were reviewed and the decision was made to consolidate objectives 2 and 3 as well as objectives 5 and 6. Objectives 1 and 4 were discusses and significantly amended. A new objective was constructed to address the need for research investigating forces and circumstances that prevent preconditioning and other practices at the cow/calf level that are known to drastically reduce BRD occurrence at the feedlot level. This objective was designed to address systems and epidemiology of BRD. In the revisions, statements to directly address topics of vaccination, judicious antimicrobial use, genetics, and environment were added.

New and revised objectives were compiled and will be emailed to the full committee for review, comment, and revision.

12:00-1:30: Lunch Break

1:30-4:30: Station Reports from attending committee members including reports from:

South Dakota-Chris Chase

California-Terry Lehenbauer

Georgia-Amelia Woolums

Mississippi-Dave Smith

Missouri-Brian Vander Ley

Virginia Tech-Tom Inzana

4:30-5:00: Business Meeting

Because of an institutional promotion, the secretary position was vacant. Dr. Vander Ley volunteered to fill both the roles of Chair and Secretary until the meeting in 2016. Drs. Vander Ley and Chase will continue to serve on the renewal writing committee.

A list of action items was developed for committee members desiring to participate in the new project.

Review, revise, and comment on objectives by October 1, 2015.

Contact your experiment station director and begin the process of completing the appendix E. Invitations should be sent automatically by email to existing members of NC-1192 requesting participation.

Contact potential new members of the committee and encourage them to participate in the new project.

Generate a word document detailing the materials and methods relating to objectives you plan to work on. Please turn send this to Dr. Vander Ley by November 1, 2015.

Accomplishments

Objective 1: To aid the rapid identification and subsequent management of BRD by developing, validating and guiding the application of new state-of-the-art diagnostic tools. University of Georgia Comparison of 4 methods to identify respiratory pathogens in dairy calves with acute undifferentiated BRD.  In collaboration with CA, we recently completed a project to determine the sensitivity of the nasal swab (NS), deep guarded nasopharyngeal swab (NPS), and bronchoalveolar lavage (BAL), as compared to the gold standard of transtracheal aspirate (TTA), for the diagnosis of viral and bacterial respiratory pathogens in dairy calves with acute undifferentiated BRD.  One hundred dairy calves with undifferentiated BRD were sampled sequentially by NS, NPS, TTA, and BAL. Calves that had ever been treated for respiratory disease, or that had received intranasal modified live viral respiratory vaccine in the previous 30 days, were excluded.  Samples were submitted for aerobic culture to identify M. haemolytica, P. multocida, H. somni, and Mycoplasma sp., and RT-PCR to identify BHV-1, BRSV, BVDV, and coronavirus.  Mycoplasma sp. isolates were tested by PCR to determine if they were M. bovis.  Agreement between the TTA and the NS, NPS, or BAL was determined by calculation of the kappa statistic.  Values of kappa were categorized according to these levels of agreement: 0.20 = poor; 0.21 – 0.40 = fair; 0.41 – 0.6 = moderate; 0.61 – 0.80 = good, and 0.81 – 1.00 = very good.  One hundred calves were enrolled.  Relative to the TTA, all sampling methods showed very good agreement for identification of Pasteurella multocida or Mannheimia haemolytica.  In contrast, for identification of bovine respiratory syncytial virus and relative to TTA, agreement was moderate for NS, good for NPS, and very good for BAL.  For identification of BCV and relative to TTA, agreement was moderate for the NS and NPS, and good for BAL.  Agreement between TTA and other sampling methods differed for different pathogens; results of BAL agreed best with the TTA for all pathogens.  Oklahoma State University Enteric Disease in Post –Weaned Calves Associated with a Recently Identified Clade of Bovine Coronavirus. Bovine coronavirus (BoCV) infections are associated with varied clinical presentations including neonatal diarrhea, winter dysentery in dairy cattle, and respiratory disease in various ages of cattle. This report presents information on BoCV infections associated with enteric disease of postweaned beef cattle in our region. In three separate accessions, one in 2012 and two in 2013 calves were observed with bloody diarrhea. One herd had the clinic cases in both 2012 and 2013. One calf in 2012 died and was necropsied and two calves from this herd died in 2013 and were necropsied. A third calf from another herd died and was necropsied. The gross and histopathologic diagnosis was acute, hemorrhagic colitis in all four cattle. Colonic tissues from all four animals were positive by fluorescent antibody and/or immunohistochemistry for BoCV antigen. BoCV was isolated in human rectal tumor cells from swabs of colon surfaces of all animals. The genomic information from a region of the S envelope region revealed BoCV clade 2. Detection of BoCV clade 2 in beef cattle in our region is consistent with recovery of BoCV clade 2 from the respiratory tract of postweaned beef calves that had respiratory disease signs or were healthy. Further investigations on the ecology of BoCV in cattle are important as the BoCV may be an emerging disease beyond the initial descriptions. Challenge studies are important to determine pathogenicity of these strains. Also there are implications to determine if current BoCV vaccines are efficacious against the BoCV clade 2 strains.   Diverse Outcomes of Bovine Viral Diarrhea Virus Infections in a Herd Naturally Infected During Pregnancy- A Case Study. A beef producer purchased Angus crossbred cattle that were pregnant with nursing calves. The purchased cattle, their nursing calves, and subsequent born calves were not initially tested for BVDV.  Bovine viral diarrhea virus subtype 2a (BVDV2a) was isolated from an aborted bovine fetus, 6.5 months,  with multiple congenital malformations including arthrogryposis, kyphosis, scoliosis, polydactylism, and  cardiac over riding aorta. Testing by immunohistochemistry and virus isolation resulted in the detection of a persistently infected (PI) yearling cohort, and a PI calf born during the same calving season. The viruses isolated from the fetus, the yearling cohort, and a PI calf born during the same calving season were identical. These malformations observed in the fetus were similar to arthrogryposis multiplexa (AM) and contractural arachnodactyly (CA) diseases with association with genetic defects in the Angus breed. The fetal tissue of the malformed fetus was negative for genetic material for AM and CA. The owner had purchased cattle that were pregnant and nursing calves, but were not tested for BVDV and with an unknown vaccination history. This case illustrates that suspect malformations should also be tested for BVDV. Also the case underscores the potential for disease after failed or inadequate biosecurity.   Bovine Herpesvirus-1: Evaluation of Genetic Diversity of Subtypes Derived from Field Strains of Varied Clinical Syndromes and their Relationship to Vaccine Strains. Bovine herpesvirus-1 (BoHV-1) causes significant disease in cattle. Control programs in North America incorporate vaccination with modified live viral (MLV) or killed (KV) vaccine.  BoHV-1 strains are isolated from diseased animals or fetuses after vaccination. There are markers for differentiating MLV from field strains using whole-genome sequencing and analysis identifying single nucleotide polymorphisms (SNPs). Using multiple primer sets and sequencing of products permits association of BoHV-1 isolates with vaccines. To determine association between vaccine virus and strains isolated from clinical cases following vaccination, we analyzed 12 BoHV-1 isolates from animals with various clinical syndromes; 9 corresponded to BoHV-1.1 respiratory group. The remaining three corresponded to BoHV-1.2b, typically found in genital tracts of cattle.  Four BoHV-1 isolates were identical to a vaccine strain; three were from post-vaccination abortion episodes with typical herpetic lesions whose dams had received MLV vaccine during pregnancy, and one from a heifer given a related MLV vaccine; Sequences of   two respiratory isolates perfectly matched mutations characterizing RLB106 strain, a temperature sensitive mutant used in intranasal and parenteral vaccines. The last three respiratory strains clearly appeared related group of MLV vaccines. Previously the MLV vaccines were grouped into four groups based on SNPs patterns. In contrast with  above-mentioned isolates that  closely matched  SNP patterns of their respective MLV vaccine virus, these 3 strains both lacked some and possessed a number of additional mutations  compared to  a group  of MLV vaccine viral genome. Finding BoHV-1.2b in respiratory cases indicates   focus should be given BoHV-1.2b as an emerging virus or a virus not recognized nor fully characterized in BRD.   South Dakota State University <ol> <li>Diagnostic findings: The infectious agents associated with bovine respiratory disease complex were monitored by bacterial culture, virus isolation, and fluorescent antibody techniques.</li> <li>Bacteriology: Bacterial agents isolated from bovine pneumonic lungs, tracheal swabs, and nasal swabs are as follows (July 1, 2013-June 30, 2014):</li> </ol> Organism                    Isolations <ol> <li>haemolytica            220</li> <li>multicida         163</li> <li>somnus 151</li> <li>trehalosi 45</li> </ol>   <ol start="2"> <li>Virology: Viral agents from bovine pneumonic lungs are as follows (July 1, 2013-June 30, 2014). There were 574 respiratory tract virus isolation attempts and 35 isolations:</li> </ol> Virus                Virus Isolations                 Fluorescent Antibody BVDV -NCP                13                    BVDV-CP                   12 BHV-1                         2                                             BRSV                                                  9 positive BHV-4 (DN-599)                     12   <ol start="3"> <li>Individual Serum Ear notch-Immunohistochemistry (IHC) or ELISA: ELISA ear notch BVDV tests were conducted on 6770 samples and there were 61 positives (0.9%).</li> <li>Molecular Diagnostics: PCR tests were done for BVDV, BHV-1, BCV, BRSV and Mycoplasma. Pooled ear notch was done on 1424 submissions with 75 positive samples, 5.3% case positive.  BVDV PCR was also done on nasal swabs, tissue, whole blood/serum /milk samples (223 submissions 18 positives, 8.0% positive).  BHV-1 PCR was also done on nasal swabs, tissue, whole blood/serum (224 submissions 11 positive).  BCV PCR was also done on nasal swabs and tissue (345 submissions 90 positive, 26%).  BRSV PCR was also done on nasal swabs and tissue (345 submissions 56 positive 16%).  There was 1 Mycoplasma bovis PCR positive samples (25 of 52, 48%).</li> <li>Characterization of BHV-1 Field isolates. Sequencing of reproductive isolates indicated that they are almost exclusively vaccine strains.  Case Histories Case 1 occurred in a herd of 106 first-calf beef heifers. The animals were vaccinated twice with Vaccine A, a MLV vaccine containing BHV-1, once at weaning in the fall of 2012 and again in February 2013. The animals were bred in June 2013 and confirmed pregnant by rectal palpation. The animals were vaccinated with an oil adjuvanted scours vaccine and treated with an anthelmintic in January 2014. Five days later the animals began to abort. Case 2 occurred in a herd of ~100 Holstein heifers. The normal vaccination protocol was animals were vaccinated with 2 to 3 doses of MLV vaccine prior to breeding and another dose of MLV was given at pregnancy confirmation. In this group animals had not been pregnancy checked for over 8 months so no vaccine had been administered to these animals in 8-12 months. The entire herd was vaccinated with either Vaccine B or Vaccine C; both vaccines were MLV vaccines that contained BHV-1. Two weeks post vaccination 8 abortions were found in the pen. Case 3 occurred in a herd of beef cows with a poor vaccination history. The animals received a single dose of inactivated multivalent viral vaccine containing BHV-1 as heifer calves. Annual revaccination with the same inactivated vaccine was infrequent. In August 2014, calves nursing these cows were given a single dose of MLV multivalent Vaccine C containing BHV-1. Beginning in January 2015, the cows began aborting.  Case 4 occurred in a herd of 350 crossbred beef cattle.  These cattle were vaccinated by a veterinarian following the manufacturer’s instructions (i.e., the cows had all been vaccinated a year prior with a similar product from the same company; heifers vaccinated twice with similar product prior to breeding). These animals were at between 5-6 months gestation and had been vaccinated with Vaccine C about 15-20 days prior to the abortions occurring. Three 3 aborted fetuses from heifers and 1 mummified fetus from a cow were recovered over the course of 2 weeks.  Results Case 1: Forty-four of the 106 calves were aborted, delivered dead, or were born alive but died within the first 24 hours. BHV-1 was isolated from 4 of the fetuses on two separate submissions. The viruses were submitted for genetic analysis to the Animal Disease Research and Diagnostic laboratory at SDSU. Single nucleotide polymorphisms (SNP) analysis indicated that the SNP pattern of the viruses isolated from the fetuses was identical to Vaccine A. Case 2: Two fetuses were submitted and virus isolation and genetic SNP analysis was performed. The SNP pattern indicated that the SNP pattern of the two isolates were identical with Vaccine C. Case 3: Two fetuses were submitted and virus isolation and genetic SNP analysis was performed. The SNP patterns of the two isolates were identical with Vaccine C.  Case 4: All fetuses from the heifers had typical histological lesions of herpesviral abortion (multiple foci of necrosis in multiple organs, some with herpetic inclusions). 3/3 were FA positive for BHV-1, while virus was isolated from 2/3. The mummified fetus was a typical mummified fetus—tissues were too autolyzed for meaningful histopathology, and all viral assays for BHV-1 were negative (FA and VI negative). One isolate was submitted for genetic SNP analysis. The SNP pattern of the isolate was identical with Vaccine C. </li> </ol> Virginia Tech Novel diagnostic tests to detect the presence of H. somni DNA in clinical samples without PCR and to detect the presence of antibodies to the biofilm EPS have been developed.  Details of this work have been presented in previous reports and have now been published or are in preparation for publication. University of Wisconsin The Wisconsin Veterinary Diagnostic Laboratory (WVDL) Virology Service (Dr. Kurth) evaluated 737 samples for respiratory viruses and mycoplasmas (Table 1 below). The greatest percentage of positive were for Mycoplasma bovis (21.2%) or Bovine respiratory corona virus (15.9%). The Bacteriology Service (Dr. Okwumabua) evaluated 1173 samples (Table 2 below). The greatest percentage of positive cultures were for Pasteurella multocida (15.8%) or Mannheimia haemolytica (11.7%). <table> <tbody> <tr> <td width="140"> Table 1 Real time PCR positive samples for viruses and Mycoplasma (737 submissions) Respiratory Pathogen </td> <td width="140"> No. Positive </td> <td width="140"> % Positive </td> <td width="140"> Change from 2013 </td> </tr> <tr> <td width="140"> Bovine respiratory syncytial virus </td> <td width="140"> 59 </td> <td width="140"> 8.0 </td> <td width="140"> Increase of 0.7% </td> </tr> <tr> <td width="140"> Bovine respiratory corona virus </td> <td width="140"> 117 </td> <td width="140"> 15.9 </td> <td width="140"> Decrease of 5.0% </td> </tr> <tr> <td width="140"> Bovine viral diarrhea virus </td> <td width="140"> 16 </td> <td width="140"> 2.2 </td> <td width="140"> Increase of 0.7% </td> </tr> <tr> <td width="140"> Bovine herpes virus 1 </td> <td width="140"> 10 </td> <td width="140"> 1.4 </td> <td width="140"> Decrease of 2.5% </td> </tr> <tr> <td width="140"> Mycoplasma bovis </td> <td width="140"> 156 </td> <td width="140"> 21.2 </td> <td width="140"> Increase of 17% </td> </tr> </tbody> </table>   Objective 2: To elucidate key steps in the dynamic interactions between pathogens, host immunity and the environment, and to determine how manipulation of these factors can reduce the risk of BRD. University of Georgia Comparison of 4 methods to identify respiratory pathogens in dairy calves with acute undifferentiated BRD.  In collaboration with CA, we recently completed a project to determine the sensitivity of the nasal swab (NS), deep guarded nasopharyngeal swab (NPS), and bronchoalveolar lavage (BAL), as compared to the gold standard of transtracheal aspirate (TTA), for the diagnosis of viral and bacterial respiratory pathogens in dairy calves with acute undifferentiated BRD.  One hundred dairy calves with undifferentiated BRD were sampled sequentially by NS, NPS, TTA, and BAL. Calves that had ever been treated for respiratory disease, or that had received intranasal modified live viral respiratory vaccine in the previous 30 days, were excluded.  Samples were submitted for aerobic culture to identify M. haemolytica, P. multocida, H. somni, and Mycoplasma sp., and RT-PCR to identify BHV-1, BRSV, BVDV, and coronavirus.  Mycoplasma sp. isolates were tested by PCR to determine if they were M. bovis.  Agreement between the TTA and the NS, NPS, or BAL was determined by calculation of the kappa statistic.  Values of kappa were categorized according to these levels of agreement: 0.20 = poor; 0.21 – 0.40 = fair; 0.41 – 0.6 = moderate; 0.61 – 0.80 = good, and 0.81 – 1.00 = very good.  One hundred calves were enrolled.  Relative to the TTA, all sampling methods showed very good agreement for identification of Pasteurella multocida or Mannheimia haemolytica.  In contrast, for identification of bovine respiratory syncytial virus and relative to TTA, agreement was moderate for NS, good for NPS, and very good for BAL.  For identification of BCV and relative to TTA, agreement was moderate for the NS and NPS, and good for BAL.  Agreement between TTA and other sampling methods differed for different pathogens; results of BAL agreed best with the TTA for all pathogens.  Acute infection with BVDV of low or high virulence leads to depletion and redistribution of WC1+ gamma delta cells ( in lymphoid tissues of beef calves.  Previous studies have shown that T lymphocytes proliferate significantly in cattle following infection or vaccination with BVDV.  However, the number, distribution, and function of these cells in the response to BVDV infection are not completely understood.   The objective of this study was to compare the abundance and distribution of T lymphocytes in lymphoid tissue during acute infection with ncp BVDV of low or high virulence.  Spleen and mesenteric lymph node (MLN) were collected from cattle at postmortem on day 5 after they were inoculated with either BVDV-1a SD-1 (low virulence; LV), BVDV-2 1373 (high virulence; HV) or control exposure (sham exposure with tissue culture medium) A higher proportion of calves challenged with BVDV showed apoptosis and cytophagy in MLN and spleen, relative to controls.  A significantly lower number of T cells was observed in spleen and MLN from calves in HV and LV groups than in controls.  Acute infection with HV or LV BVDV resulted in depletion of WC1+ T cells in mucosal and systemic lymphoid tissues at 5 days after challenge.  Oklahoma State University Pulmonary Lesions and Clinical Disease in Calves Challenged with Histophilus somni Five Days after Tildipirosin or Tulathromycin Treatment. The objective of this study was to compare in calves the efficacy of two macrolide antimicrobials, tildipirosin and tulathromycin, in a metaphylaxis model of experimentally induced Histophilus somni respiratory infection. Twenty-four Holstein and Holstein - crossbreed steers, approximately 4 months of age that had low to no anti-H. somni antibodies were enrolled in the study. On day 0, 8 calves each were treated by subcutaneous injection:  Group 1 - tildipirosin   at 4 mg/kg, Group 2 - tulathromycin at 2.5 mg/kg, and Group 3 (Control) - isotonic saline. On day 5, all calves were inoculated intrabronchially with 1.6 x 109 CFU/ml of H. somni strain 7735. Clinical evaluations were done on days 5-8. On day 8, calves were humanely killed, and lungs evaluated for pneumonia and bacterial isolation.  Twelve hours after challenge, clinical scores for each group significantly increased compared to scores at challenge. On days 6, 7, and 8, clinical scores for the tildipirosin-treated group were significantly lower compared to the tulathromycin-treated and Control calves. On days 6 and 7, clinical scores for tulathromycin-treated calves were significantly lower than for the Control calves. Percentage of pneumonic lung was significantly lower for the tildipirosin-treated group (6.0 ± 2.3) than for tulathromycin (30.9 ± 21.5%) and Control groups (26.3 ± 17.6%). H. somni was reisolated from tulathromycin-treated and Control groups. Within the parameters of this metaphylaxis model, tildipirosin was generally more efficacious than tulathromycin in minimizing clinical disease. Objective 3: To investigate the mechanisms by which infectious agents work singly or in combination to evade, suppress, or misdirect the host immune response, or to directly induce cellular or molecular pathology, in BRD.   South Dakota State University Neutrophils are the predominant white blood cells in peripheral blood activate the innate as well as adaptive immune response. In the current study, the effect of bovine viral diarrhea virus (BVDV) infection viability, surface marker expression such as CD14, CD18 and L-selectin, neutrophil’s migration and phagocytosis ability was investigated. Isolated neutrophils (CD14+, CD18+ and L-selectin+.) were infected with homologues pair of BVDV [e.g. cytopathic (cp) BVDV1b-TGAC or non- cytopathic (ncp) BVDV1b-TGAN] at 3 MOI.  Neutrophils were examined for apoptosis and CD14, CD18 or L-selectin expression at 1 hr and 6 hrs post infection (PI). Neutrophil migration was measured through transwell assay with BVDV or LPS treated bovine macrophages in lower chamber and freshly collected neutrophils in the upper chamber. To measure the effect of BVDV on phagocytosis ability of neutrophils, neutrophils were infected with cpBVDV1b-TGAC or ncpBVDV1b-TGAN at 3 M.O.I for one hour and phagocytosis ability was measured through 0.2µm florescent beats. Both biotypes of BVDV induced the apoptosis in neutrophils. The cp BVDV1b-TGAC induced more apoptosis than its homologues ncp BVDV1b-TGAN at 1 and 6 hrs PI. Similarly, both biotypes and LPS down regulated the L-selectin expression on bovine neutrophils with course of infection. The LPS up regulated the CD14 expression while down regulates the CD18 expression on neutrophils. Whereas both biotypes of BVDV down regulated the CD14 expression and upregulate the CD18 expression on bovine neutrophils. BVDV infection significantly reduced the neutrophils migration and phagocytosis ability (p < 0.05). The cp BVDV1b-TGAC significantly reduced neutrophil’s migration and phagocytosis ability as compare to its homologues ncp BVDV1b-TGAN (p <0.05). Virginia Tech The biofilm formed by P. multocida has been characterized. Strains that are the most highly encapsulated, and are often isolated from acute cases of bovine respiratory disease, produce the most capsule, but make the least amount of, most rough, and least dense biofilm.  However, even highly encapsulated, poor biofilm forming strains can reside in an H. somni biofilm.  After 2-3 days incubation, H. somni is difficult to isolate and P. multocida becomes prominent or the sole agent of the biofilm. The amount of polysaccharide also diminishes over time in P. multocida mono-biofilms or in polymicrobial biofilms, but continues to increase in H. somni mono-biofilms. Objective 4: To develop management practices, including rationally applied therapeutic and preventative interventions that minimize the impact of BRD on cattle health, welfare and productivity University of Georgia Evaluation of the effect of an injectable trace mineral (ITM) supplement on the immune response of dairy calves to viral vaccines.  Previous studies demonstrated that administering injectable trace minerals (ITM; Zn, Mn, Cu, and Se) improved humoral immune response to BHV-1 following administration of a MLV vaccine in cattle (Arthington and Havenga, 2012). The objective of this study was to evaluate the effect of an ITM supplement containing zinc, manganese, selenium, and copper on the humoral and cell mediated immune (CMI) responses to individual vaccine antigens in dairy calves receiving a modified-live viral (MLV) vaccine containing BVDV, BHV1, PI3V and BRSV. Thirty dairy calves (3 months of age) were administered 2 mL of a 5-way MLV vaccine and 2 mL of an attenuated-live M. haemolytica and P. multocida bacterin subcutaneously (SC). Calves were randomly assigned to 1 of 2 groups:  (1) Subcutaneous administration of ITM (1 mL/100 lb BW, ITM; MultiMin 90, Fort Collins, CO; n = 15) or (2) Subcutaneous injection of sterile saline (2 mL, Control; n = 15). Three weeks after initial vaccination, calves received a booster of 2 mL of the 5-way MLV vaccine, and 2 mL of the attenuated-live bacterin SC. Concurrently with the vaccine booster, a second administration of injectable trace minerals or sterile saline SC was given to calves in ITM and control group, respectively. Blood samples were collected on days 0, 7, 14, 21, 28, 42, 56, and 90 relative to prime vaccination for antibody titer determination, antigen-induced in vitro IFN-γ production by peripheral blood mononuclear cell (PBMC), and antigen-induced PBMC proliferation.   Administration of ITM concurrently with MLV vaccination resulted in higher antibody titer to BVDV-1 on day 28 post prime vaccination compared to the control group (P=0.03). There was a tendency of a higher PBMC proliferation response to BVDV (P=0.08) on day 14 post prime vaccination in calves treated with ITM than in the control group. Additionally, calves treated with ITM showed an earlier and more consistently increased PBMC proliferation to BVDV following MLV vaccination (on days 14, 21, and 42 relative to day 0), compared to the control group (only on day 28). There was a significantly higher PBMC proliferation upon BRSV stimulation on day 7 post prime vaccination in the ITM group than the control group (P=0.01). Calves treated with ITM showed a significantly augmented PBMC proliferation upon stimulation with BRSV on days 7, 14, and 42 after prime vaccination relative to day 0 (P <0.05). Proliferation of PBMC was increased upon BHV1 recall in both ITM and control groups on days 14, 21 28 and 42 post vaccination compared with day 0 (P<0.05). Significant differences were not found in the production of IFN-γ by PBMC after stimulation with BVDV, BHV1, and BRSV between calves treated or not with ITM. In conclusion, administration of ITM concurrently with vaccination in dairy calves resulted in increased antibody titer to BVDV1 (on day 28 after prime vaccination) and PBMC proliferation after BVDV and BRSV stimulation compared to the control group. Case-control study of herd-level and calf-level risk factors for nursing (preweaning) calf BRD in cow-calf herds.  In collaboration with KS, SD, ND, and MS, we have completed a case-control study of herd-level risk factors for pneumonia in nursing beef calves.  Herds were enrolled in NE, SD, and ND. Case herds treated at least 5% of their nursing calves for BRD, while control herds selected randomly from the same veterinary practice treated no more than 0.5% of their nursing calves for BRD.  A total of 84 herds were enrolled in the study: 30 case herds and 54 matched control herds.  Fifty-two of the herds were enrolled in 2012, 24 were enrolled in 2013, and eight were enrolled in 2014.  Twenty-nine of the herds were located in Nebraska, 23 were located in North Dakota, and 32 were located in South Dakota.  In the multivariable analysis, three variables were significantly associated with calf BRD: herd size; the use of intensive grazing; and synchronizing cows and heifers after calving.  Compared to herds with fewer than 150 cows, the odds of having > 5% incidence of calf BRD were 7.9 times higher for herds with 150-499 cows, and 12 times higher for herds with 500 cows or more.  Compared to herds that did not use intensive grazing, the odds of having > 5% incidence of calf BRD were 3.3 times higher for herds that did use intensive grazing.  And compared to herds that did not use a synchronization program after calving, the odds of having > 5% incidence of calf BRD were 4.5 times higher for herds that did use a synchronization program. Immune response to subcutaneous and intranasal vaccination in young beef calves.  In this study we compared the serum neutralizing antibody (SNA) titers to BHV1 and BRSV and nasal mucosal BHV1-specific IgA levels following IN or SC MLV booster 60 days after IN MLV priming in young beef calves. The objective was to determine whether IN or SC booster following IN priming induced different immune responses.  We used 24 Angus calves (1-3 weeks of age) that received an IN prime vaccine containing BHV1, BRSV and PI3 (Inforce-3®) and 60 days later were boostered with 2ml of the same vaccine either IN (n=12) or SC (n=12). Calves had high SNA titers to BRSV and BHV1 at 1-3 weeks of age, likely due to maternal antibody.  A significant decrease in SNA titers to BRSV and BHV1 was observed 2 and 8 weeks after IN prime vaccination. Booster vaccination 60 days after priming (IN or SC) did not cause increase in SNA titers against BHV1. Intranasal booster vaccination induced a significant SNA response to BSRV 2-4 weeks after booster. On the other hand, calves that received SC booster vaccine did not show increase in SNA to BRSV.   A sustained increase in BHV1 specific IgA titers in nasal secretions was observed in both (IN & SC) groups post prime and booster vaccination.   By comparing the fold change in IgA in nasal secretions on day 21 relative to the day of booster vaccination, two patterns of response were observed in the calves that received IN booster. The calves with strongly enhanced nasal IgA to BHV1 after priming did not show clear recall response to IN booster vaccine, but those with low nasal IgA to BHV1 at the time of booster showed a strong (≥8 fold changes) recall titer on 3 or 4 weeks post IN booster.  In contrast, a significant BHV1 nasal IgA response (>8 fold increase) was observed in six calves that received a SC booster regardless the level of BHV1-specific IgA in nasal secretions by the time of booster. In summary, in this study calves primed by IN vaccination had significantly higher SNA titers to BRSV following IN booster, but not SC booster.  In contrast, BHV1 SNA titers did not increase following either IN or SC booster. However, nasal BHV1-specific IgA concentration was significantly increased following IN booster of calves if their IgA titers after priming were not high; SC booster also increased nasal BHV1-specific IgA titers.  The results of this study, which is scheduled to be repeated in fall 2015, suggest that booster route influences the immune response in primed cattle, but the response may not be the same for all pathogens.  More work will be needed to clarify the significance of these results.  University of Missouri We, along with collaborators from the University of California-Davis and Auburn University, have determined that orally administered iodine may be used in the respiratory tract of cattle to inactivate pathogens commonly associated with bovine respiratory disease. With further research, these findings may translate into alternative treatment and preventative therapies that can reduce the use of conventional antimicrobials. Mississippi State University Evaluation of the effect of on-arrival vaccination and deworming on stocker cattle health and growth performance:  High risk stocker cattle are commonly vaccinated and treated with anthelmintics at arrival; however, little research has been published which provides evidence that these practices are beneficial.  It is possible that vaccination of cattle that may be ill, such as some newly arrived high risk cattle, could be beneficial.  We wished to evaluate the effect of two common management practices, vaccination and deworming, on health and growth of high risk stocker cattle over an 84-day conditioning period.  Sera were also collected for measurement of SN titers to BHV-1, BRSV, and BVDV at multiple points over time; those results will be reported later.  Auction market derived calves (n=80) received from an order-buyer were stratified by d-3 weight and fecal egg count into 20 pens of 4 animals each. Pens were randomly assigned to treatments in a 2x2 factorial design to test vaccination at arrival (d0 modified-live BRD and clostridial vaccine or not) and deworming (d0 oral fenbendazole and levamisole or not). Body weight and blood was collected days 0, 14, 28, 42, 56, 70 and 85.  Fecal egg counts were measured days -3, 28, 56, and 85. Clinical signs of BRD were monitored daily. Treatment effects on BRD incidence, mortality, and growth were tested using Poisson, logistic, or linear regression, respectively (α≤0.05). BRD incidence was greater for calves with d0 vaccination (RR=3.2), high (≥104F) fever at day 0 and higher d-3 FEC. Mortality was greater for d0 vaccination and high fever.  Growth was lower for d0 vaccination (-10.3 lbs), moderate (103-103.9F) and high fever (-24.1 lbs and -16 lbs, respectively), and number of times treated for BRD (-17.5 lbs/treatment). Deworming at arrival decreased fecal egg counts in treated cattle but was not found to be significantly associated with BRD morbidity, mortality, or weight gain. South Dakota State University The Genetics of Feedlot Health This station along with Texas, Illinois, Missouri, New York and Colorado are involved in the Genetics of Feedlot Health Project the project was performed in 2009 and 2010.  This study looks at behavior, genetics, nutrition along with microbiology and immunology on respiratory disease and carcass quality. We have continued to analyze the immunological data and are analyzing cortisol and it relationship to overall antibody level and proinflammatory responses. Objective 5: To promote open scientific exchange and dialogue among scientists, veterinarians, allied industry professionals and cattlemen to advance BRD research initiatives. University of Georgia With SD and CA, GA worked with the publisher of Animal Health Research Reviews to publish the proceedings of the 2014 BRD Symposium.  This means that the Symposium proceedings are now identifiable by searches on PubMed, and anyone at an institution with a subscription to the journal will be able to access the papers.  Thus the information presented at the 2014 BRD Symposium will be widely available.  University of Missouri Our findings have been presented at the Merial Veterinary Research Scholars Symposium and have been accepted for presentation at the annual Conference of Research Workers in Animal Diseases. In addition, a manuscript detailing our findings is being prepared. South Dakota State University The 2014 BRD symposium was held in Denver CO in July-August 2014.  With attendance of over 300 including producers, veterinarians, government officials and animal health industry professionals, the meeting was a success.   The 6th BVDV-Pestivirus meeting held in October 2015 in Kansas City MO.  This international conference had over 125 attendees. SD provided BRSV isolates toCA & GA and BHV-1 isolates to OK. Objective 6: To facilitate the translation of research findings to practical field application by developing and integrating BRD educational programming for national veterinary and producer organizations focused on cattle health and management. Oklahoma State University Impact of Species and Subgenotypes of Bovine Viral Diarrhea Viruses on Control by Vaccination.   Bovine viral diarrhea viruses (BVDV) are diverse genetically and antigenically. This diversity impacts both diagnostic testing and vaccination. In North America there are two BVDV species, 1 and 2 with 3 subgentoypes, BVDV1a, BVDV1b and BVDV2a. Initially U.S. vaccines contained BVDV1a cytopathic (CP) strains. With the reporting of BVDV2 severe disease in Canada and the U.S. there was focus on protection by BVDV1a vaccines on BVDV2 disease. Also emphasis of controlling persistently infected (PI) cattle resulted in studies for fetal protection afforded by BVDV1a vaccines. Initially, studies indicated that some BVDV1a vaccines gave less than 100% protection against BVDV2 challenge for fetal infection. Eventually vaccines in North America added BVDV2a to modified live virus (MLV) and killed BVDV1a vaccines. Ideally vaccines should stimulate complete immunity providing 100% protection against disease, viremias, shedding, and 100% fetal protection in vaccinates when challenged with a range of diverse antigenic viruses (subgenotypes).  There should be a long duration of immunity stimulated by vaccines, especially for fetal protection. MLV vaccines should be safe when given according to the label and free of other pathogens. While vaccines have now included BVDV1a and BVDV2a, the discovery of the predominate subgenotype of BVDV in the U.S. to be BVDV1b, approximately 75% or greater in prevalence, protection in acute challenge and fetal protection studies became more apparent for BVDV1b. Thus many published studies examined protection by BVDV1a and BVDV2a vaccines against BVDV1b in acute challenge and fetal protection studies. There are no current BVDV1b vaccines in the U.S. There are now more regulations on BVDV reproductive effects by the USDA Center for Veterinary Biologics (CVB) regarding label claims for protection against abortion, persistently infected calves, and fetal infections, including expectations for studies regarding those claims. Also USDA CVB has a memorandum providing guidance for exemption of the warning label statement against the use of the MLV BVDV in pregnant cows and calves nursing pregnant cows. In reviews of published studies in the U.S, the results of acute challenge and fetal protection studies are described including subgenotypes in vaccines and challenge strains and the results in vaccinates and the vaccinates fetuses/newborns. In general vaccines provide protection against heterologous strains, ranging from 100% to partial but statistically significant protection. In recent studies, the duration of immunity afforded by vaccines was investigated and reported. Issues of contamination remain, especially since fetal bovine serums may be contaminated with noncytopathic BVDV. In addition, the potential for immunosuppression by MLV vaccines exists, and new vaccines will be assessed in the future to prove those MLV components are not immunosuppressive by experimental studies. As new subgenotypes are found, the efficacy of the current vaccines should be evaluated for these new strains.   South Dakota State University Classes specific on bovine respiratory disease were taught to the advanced group of the United States Dairy Education Consortium at Clovis NM in June 2015.

Publications

Palomares RA, Sakamoto K, Walz HL, Brock KV, Hurley DJ.  Acute infection with bovine viral diarrhea virus of low or high virulence leads to depletion and redistribution of WC1+ cells in lymphoid tissue of beef calves.  Vet Immunol Immunopathol, 2015.  In press.  Palomares RA, Hurley DJ, Woolums AR, Parrish JE, Brock KV.  Analysis of mRNA expression for genes associated with regulatory T lymphocytes (CD25, FoxP3, CTLA4, and IDO) after experimental infection with bovine viral diarrhea virus of low or high virulence in beef calves.  Comp Immunol Microbiol Infect Dis. 2014. 37:331-338. Palomares RA, Parrish J, Woolums AR, Brock KV, Hurley DJ.  Expression of toll-like receptors and co-stimulatory molecules in lymphoid tissue during experimental infection of beef calves with bovine viral diarrhea virus of low and high virulence.  Vet Res Commun. 2014 38:329-335.  Klingenberg M.A., Shoemake B.M., Nolan R.A., Heller M.C., Newcomer B.W., Meyer A.M., Vander Ley B.L. “Iodine secretion in airway fluid following a single bolus of sodium iodide.” Poster for the Merial Veterinary Scholars Symposium. August 2015. Heller M.C., Clothier K.A., Newcomer B.W., and Vander Ley B.L. “Sodium iodide inactivates Mannheimia haemolytica and Bibersteinia trehalosi in vitro.” Abstract for the Conference for Research Workers in Animal Diseases. December 2015. Newcomer B.W., Vander Ley B.L., Galik P., and Heller M.C. “In vitro inactivation of bovine viral respiratory pathogens using an iodine-based antimicrobial system.” Abstract for the Conference for Research Workers in Animal Diseases. December 2015. Shoemake B.M., Vander Ley B.L., Klingenberg M.A., Nolan R.A., Meyer A.M., Schultz L.G., Newcomer B.W., and Heller M.C. “Iodine Secretion in airway surface fluid following a single oral bolus of sodium iodide in calves.” Abstract for the Conference for Research Workers in Animal Diseases. December 2015. Fulton, R.W., Rezabek, G.B., Grant, R., Ridpath. J.F., Burge.L.J.: Diverse Outcomes of Bovine Viral Diarrhea Virus Infections in a Herd Naturally Infected During Pregnancy- a Case Study.. Bovine Practitioner, 48: 95-98, 2014.   Fulton, R.W., Herd, H.R., Sorensen, N.J., Confer, A.W., Ritchey, J.W., Ridpath, J.F., Burge.L.J.: Enteric Disease in Postweaned Beef Calves Associated with  Bovine Coronavirus Clade 2. Journal of Veterinary Disease Investigation, 27: 97-101, 2015.   Fulton, R.W., d’Offay J.M., Eberle,R., Moeller, R.B., Van Campen, H., O’Toole, D., Chase, C., Miller. M.M., Sprowls, R., Nydam, D.V.: Bovine Herpesvirus-1: Evaluation of Genetic Diversity  of Subtypes Derived from Field Strains of Varied Clinical Syndromes and Their Relationship to Vaccine Strains. Vaccine, 31: 549-558, 2015.   Fulton, R.W.: Impact of Species and Subgenotypes of Bovine Viral Diarrhea Virus on Control by Vaccination. Animal Health Research Reviews, 16: 40-54, 2015.   Holbrook T, Gilliam L, Stein F, Morgan S,  Avery A, Confer AW, Panciera RJ. Lathyrus hirsutus (Caley Pea) Intoxication in a Herd of Horses. J Vet Intern Med 29 :294-298, 2015.   Taylor JD, Holland B, Step DL, Payton ME, Confer AW. Nasal isolation of Mannheimia haemolytica  and Pasteurella multocida as predictors of respiratory disease in shipped calves. Res Vet Sci 99:41-45, 2015.   Confer AW, Snider TA, Taylor JD, Montelongo M, Sorensen NJ. Pulmonary lesions and clinical disease in calves challenged with Histophilus somni five days after tildipirosin or tulathromycin treatment. AJVR, in press 2015.   Pan, Y., T. Fisher, C. Olk, and T. J. Inzana. 2014. Detection of antibodies to the biofilm exopolysaccharide of Histophilus somni following infection in cattle by enzyme-linked immunosorbent assay.  Clin. Vac. Immunol.  21:1463-1467. Shah, N., A.B. Bandara, I. Sandal, and T.J. Inzana.  2014. Natural Competence in Histophilus somni strain 2336.  Vet. Microbiol. 173:371-378.   Petruzzi, B., R.E. Briggs, W.E. Swords, C. De Castro, A. Molinaro, T. J. Inzana. 2014.  Polymicrobial Biofilm formation by Pasteurella multocida and Histophilus somni.  Abst. 14.  1st ASM Conference on Polymicrobial Infections.  Nov. 13-16, 2014. Washington, DC. Petruzzi, B.L., R.E. Briggs, C. De Castro, A. Molinaro, T. Inzana. Characterization of Biofilm Formation by Pasteurella multocida.  Oral Pres. 018. Conf. Res. Workers Anim. Dis. 2014 Ann. Meet. Dec. 7-9, 2014. Chicago, IL. Petruzzi, B., W.E. Swords, R. Briggs, and T.J. Inzana. 2015. Characterization of biofilm formation in Pasteurella multocida. Mid-Atlantic Microbial Pathogenesis Meeting 2015. Jan. 25-27, 2015.Wintergreen, VA.   Section 7. Scientific and Outreach Oral Presentations. Palomares, RA, Sakamoto, K, Waltz, H, Brock, KV, Hurley, DJ. Acute infection with        bovine viral diarrhea virus causes depletion of <a href="http://www.ncbi.nlm.nih.gov/pubmed/22238587"> WC1+ γδ T cells</a> in lymphoid tissues in     beef calves. 95th Annual CRWAD meeting, December 7-9, 2014 Chicago, IL. Abstract Woolums A.  Review of Immunity in the Calf, and Vaccination to Control BRD.  Northwest Women Veterinarians’ Meeting, Walla Walla WA.  July 29, 2015. Woolums A.  The Impact of Vaccination and Maternal Immunity on Respiratory Disease in Calves.  XV. Middle European Buiatric Congress and 10th European College of Bovine Health Management Symposium.  Maribor, Slovenia.  June 10-13, 2015.  Woolums A.  BRD Diagnostic Testing in Stocker Cattle. Meeting the Stocker Challenge.  Starkville MS.  December 9-10, 2014. Woolums A.  Bovine Respiratory Vaccinations: When, Why, and How?  University of Georgia Continuing Education Conference for Veterinary Technicians.  Athens GA.  October 18, 2014. Griffin CM; Karisch B; Woolums AR; Blanton J; Kaplan RM; Epperson W; Smith DR. Evaluation of on-arrival vaccination and deworming on stocker cattle health and growth performance.  Mississippi State University College of Veterinary Medicine Research Day, August 13, 2015.  T.J. Braud, B.B. Karisch, D.R. Smith, C.L. Huston, S.G. Genova. 2015.  Effect of crude protein levels and metaphylaxis during the stocker receiving phase on feedlot and carcass performance. ASAS Southern Section.  Smith DR. 2015. Stocker cattle receiving programs.  Mississippi Veterinary Medical Association, Summer Meeting. Orange Beach, AL. July 2015. Smith DR. 2015. Pneumonia in calves prior to weaning.  Mississippi Veterinary Medical Association, Summer Meeting. Orange Beach, AL. July 2015.  Schneider LG, Smith DR. 2015. The effect of morbidity on weaning weight of beef calves.  Conference of Research Workers in Animal Diseases.  Chicago.  Oral Presentation 081.  Dec 8, 2014. Brown, J., Woolums, A.,Jones, L,McKinnon, G., Fulton, R.W., Ridpath, J.: Investigation Into an Outbreak of Respiratory Disease in Nursing Calves in a South Georgia Beef Herd. Bovine Respiratory Disease Symposium (BRDS) 2014. Denver , CO. July 30-31,2014. Fulton, R.W.: Bovine Coronaviruses: Respiratory and Digestive Tract Infections/Disease in Post Weened Beef Calves. Academy of Veterinary Consultants Meeting. Augest 1-2,2014. Denver, CO. Herd, H.R., Ritchey, J.W., Fulton, R.W., Ridpath, J.F.: Coronavirus-Associated Hemorrhagic Colitis in Two Young Adult Beef Cattle in Oklahoma. 57th Annual Meeting of AAVLD, October 26=22-21,2014. Kansas City, MO. Fulton, R.W.: Impact of Species and Subgenotypes of BVDV on Control by Vaccination. Joint U.S. BVDV/ESVV Pestivirus Symposium. October 14-15,2014. Kansas City, MO. Confer AW, Taylor JD.  Pasteurella multocida in bovine respiratory disease: What we know and what we don’t know. Summer Conference, Academy of Veterinary Consultants, Denver, CO, 2014.  Exploring mechanisms of Histophilus somni virulence: from phase variation to biofilm formation.  2014.  University of Mississippi, College of Veterinary Medicine. Biofilm and Exopolysaccharide Production by Histophilus somni and Pasteurella multocida individually and during Polymicrobial Interaction.  2014.  European Cooperation in Science and Technology Conference on Microbial cell surface determinants of virulence as targets for new therapeutics in Cystic Fibrosis.  Università Di Napoli Federico Ii, Naples, Italy.

Impact Statements

Polymicrobial Biofilm Formation by Pasteurella multocida and Histophilus somni in vitro and in the Bovine Host. Internal Research Competition, College of Veterinary Medicine, Virginia Tech, 7/1/13-6/30/15.