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Brief Summary of Minutes

SEPTEMBER 19:


Meeting began 8:30 am. Brief discussion of applying for CAP when next opportunity arises, per discussions between Chris Chase and Amelia Woolums with Peter Johnson over e-mail. CAP award for BRD is conceivable; time frame within which we could apply depends on when next award is open. The Johnes CAP has been renewed for 4 years ($1.2 million/year); the avian flu CAP is applying for a renewal, and the PRRS CAP is submitting a revised renewal proposal. If either PRRS or AI arent renewed, funds will be freed up to begin a new CAP; if they are both renewed, a new CAP wont be possible in FY2009 (or longer).


Station reports 8:45 am - 12:20 pm with break 10:30-10:45.


Lunch (sponsored by Novartis Animal Health) began at 12:20.


1:00 pm: Teleconference with Gary Sherman. Discussed points in our letter to him of August 17th. Has talked with people at CSREES about reports from other multistate projects. Some are feeling less successful, as are we; others doing better. Some talk at CSREES re that, if many projects are having problems like us, changes may be made so that projects can function more effectively (as multistate projects are vital to CSREES mission). Wont be fast, but hes going to try to have gatherings at CSREES, maybe a conference on revitalizing multistate committee process, to find out what can be done to improve situation. Principles are the Ag Expt Station directorsCSREES is just in an overview position, as the law provides. Law very broad, so lots of discretion left to Ag Expt Station directors to use Hatch money as at they see fit. Not enough money to go around as with everywhere elsestate support for universities dropping so much has contributed a lot to this. Thus Ag Expt station directors re-direct Hatch funds to do whatever they feel is most necessary. This is allowable under law. Law does state that 25% of funds need to go to multistate projectsthis is audited to ensure compliance. But many different activities can fit under this 25%. Should talk to our Ag Expt Station director re how this is broken down. Bottom line: Expt Station directors are using their authority in difficult times to do the best they can with the funds they have. Not to say we cant make a case re BRD being an area that requires particular support&but need to keep this in mind when we visit Expt Station Directors. Also remember that BRD is still well supported by NRI, as compared to other animal diseases (tho he notes that problem with getting funds from NRI to study more applied problems is valid). --remember that other researchers are far worse off than we are, in that BRD is at least still supported by NRI.


Money going in to Hatch has been steady, but in real dollars has declined. State funding for Expt Station activities has also declined.


Every group needs to be known to their Ag Expt. Station directorsmake your case whenever necessary. More than in old days, need to make our mission more clearneed to do a bit more marketing of what we are trying to do than used to be necessary.


Have had similar discussions with APHIS and ARSthey all need more money, but they cant lobby!


One can argue that we are in perfect storm AI (avian flu), BSE, etc. do make animal health on the radar of the Secretary of Homeland Security. Gary was at meeting with aide of Sec of HS: how high on list is animal health on radar of Sec HS answer was that animal health was high --may need to make marriage of convenience with bioterrorism/agroterrorism link and BRD --we are in a time where the best chance for seeking extra funding is hereBUT the federal budget right now not in a good place due to war on terrorism.


Comment from Ricardo Rosenbusch: article from Sciencesome think industry will pick up slack in human health research funding that government doesnt supplybut a study shows that industry follows lead of federal fundingso we might expect similar outcomes for animal health. GS agreed that, from his experience, this is true.


Gary will finish going through our 6 points; answers probably wont satisfy us, but he hopes to have more information back to us in next year re challenges multistate projects are facing. Old model clearly is not workingthey cant lobby, but they can talk about how programs can best be run in current climate: how to write laws, make programs that are more supportive. Hopefully in form of conferences, etc. Especially good to try to figure out what is different between multistate projects doing well vs those not doing wellwhat lessons can we learn from successful projects?


Specific comments re points in our letter to him:


1. Hatch funds havent declined relatively, but have declined in terms of real dollars. Bigger problem: Ag Expt Station directors have made necessary redirection of funds to preserve faculty lines. He had this experience personally. University administrators must do creative financing to keep faculty positions in place. Some faculty may not realize how thinly funds are spread, and how creative administrators must be, to keep faculty lines in place. This type of use of funds is within discretion of Ag Expt Station directors.


2. Re disproportionate funding/redirection of funding: again, related to discretion of Ag Expt Station director at each institution. Gary thinks disproportionate funding is probably exactly a problem, and we can only find out reasons via frank discussions with Ag Expt Station directors. He thinks in most cases while, decisions may seem unfair/capricious, these people are actually doing the best they can with a bad situation.


3. Re question of whether funding available from our Experiment Stations merits participation in NC1027: he hopes NC1027 never goes away, but he also realizes we can only be so altruistic about where we spend our time. He hears from lots of land grant colleges who say that 75% of their funding comes from NIH, so thats what they have to work on. Related to our later point that BRD not a big model for human disease, so realizes that we cant tap into NIH funds as well as other veterinary researchers. This is probably one of the stronger arguments to make to veterinary school administrators and Ag Expt Station directors. Redistribution of Hatch funds needs to be done on a transparent basis. If we have good logic re why were not in good shape for competing for NIH money, but relative importance of field were in is high (and this is true for BRD)constantly make comparison between losses due to BRD and level of funding for BRD researchthis needs to keep being drummed on til it hits the right set of ears. Is participation in NC1027 worth it? He leaves that decision to us, but the fact that we are thinking this way gives him some incentive to move forward to get CSREES administrators to look at whether multistate projects need to be run under a different model. Some multistate projects are in worse shape than us.


4. Re this [instances when scientists at historically participating stations have wanted to participate but cant get support due to funds going to other departments/colleges]: he will look into this. Could be that Expt Station Director cant transfer funds to the dept. this person is in? We told him to call Jeff Lakritz at Ohio State for more details. R. Rosenbusch asked: can you address as 2 separate issueswhether the person can even get travel funds, vs whether the person can get research funds? We told him Jeff would probably appreciate even travel funds.


5. Re Hatch funds being last vestige of support for veterinary translational/applied researchthis is the case we need to make with our Ag Expt Station directors. Point out this gapits real. Worse for some diseases than others. Make this point to amplify our other point that there may be too few people left in this field to provide critical mass to support animal health the way it needs to. We need to continue to make this case that the gap is real between what is fundable at NRI vs what we need so that it doesnt dead-end at basic research level.


6. Re brain drain from BRD researchhe asked how many grad students are here at our meeting? We told him none. This should be a major arrow in our quiver why we need our share of funds from our respective Expt Stations. Reality is that we are in a time when we all need to market better what we need for what we are doing.


Other comments from Gary Sherman: Strategies for revitalizing multistate projects: some have done well via meeting at CRWAD. They see rationale for going to AABP, but in the sense of trying to get more research money, and seeing blend of basic and applied research, CRWAD is good venue. PRRS had a conference and had great turnout; brucellosis group did too. A BRD symposium at CRWAD might be a good idea.


Robert Fulton: how much should we emphasize clinically applied research with Hatch/1433 money? They are encouraged at their station to use Hatch/1433 to serve as springboard for another grant to an outside agency. Which do you think Hatch/1433 funds are supposed to be used for? G. Sherman says he hears people above him say that Hatch/1433 should be used to solve practical problems, but reality is probably that your institution needs you to use the funds as springboard to get more extramural funds. However, they do say these funds are supposed to be for local/applied problems, so realizes they are speaking out of both sides of their mouth. Again, problem is that there is just not enough money to do both. Hatch/1433 funds are not enough to support a research program any morethus reality is that people are using these funds to get more extramural funds.


Ricardo Rosenbusch: is there any merit in bringing these attentions to AABP, AVC, producer groups? Although as a USDA employee G. Sherman cannot be involved with lobbying, he acknowledged that it could be helpful for any group in our position to get organized, and then ask to meet with Secretary Johanns [who resigned the morning after this discussion] and others. Stakeholder groups important from political standpoint; CRWAD important from a scientific standpoint. We need to be focused on both arms of this: pushing for political attention, and pushing to show scientific strengths of our group. Some groups that are effective have a governmental relations committee of their own, focused like a laser, on bringing these problems to the attention of people in government. Gale Buchanan (former head of Dept. of Animal and Crop Sciences at University of Georgia) is our undersecretary.


Christopher Chase comment: we dont have grassroots support like CRWAD, so this is why we are meeting at AABPuntil we get the political mandate that the PRRS group does, meeting at CRWAD may not do us as much good. G. Sherman concedes that if we dont have enough manpower to do both, we need to get constituent groups organized to help us bring attention to the problem first.


2:20 - 3:15 pm: Discussion of possible symposium on animal health research funding. A. Woolums presented proposed agenda, list of invitees, objectives/outcomes. Also presented 2 other options: a) BRD symposium to show current research, and have discussions, or b) just write white paper and take it out to groups. We planned to submit conference grant to NRI to support this, but P. Johnson said NRI cannot support conference if it is explicitly aimed at lobbying. Could support a national/international conference on BRD research, which could be helpful to showcase state of research and needs for future research.


Group discussion: BRD symposium would be more positiveeasy to selleasier to get funding. This is sounding like better outcome. We were successful in 2001 getting support for AVC symposium. Comment: meeting at Amarillo in 1983 had better stature. Proceedings were referenced for years. Comment: Ray Loan and Texas A&M really got behind that meeting. AVC supported with attendees. Texas Cattle Feeders supported banquet, got behind meeting politically. If we can get Texas Cattle Feeders behind us, good year-to-year support. Consider American Farm Bureau, too.


Two years from now: AABP in Omahamight be good site. Or Albuquerque (both AVC and AABP meeting there, but in 3 years). AVC more diversemore cow-calf, some dairynot just feedlot, like is thought. Consensus was that would be ideal to have symposium a day or two before AVC. If we apply for NRI conference grant and are funded, would get money fall 08; need to spend by fall 09. Committee to write grant and put together plan for BRD Research Symposium: A. Woolums (chair), C. Chase, R. Fulton, R. Rosenbusch. Will model on Amarillo 2003 BRD meeting; tentatively plan to hold in conjunction with AVC at a centrally located U.S. site. Comment: Animal Health Research Reviews might be good site for proceedings to be published. Can get electronically.


Short break 3:15-3:20.


3:20- 3:25. Short presentation by LA rep; Dr. Navarre here for Dr. Kousoulas, and couldnt attend this morning

3:25- Discussion with members of biologics/pharmaceutical industry on BRD research funding and ways to optimize interaction between industry/researchers outside industry, chaired by Dan Grooms. Attendees: Dale Groeteleuchen from Pfizer, Bruce Nosky, Merial; Richard Harlan, Novartis; Gerry Mechor, Elanco. Highlights: remember industry needs to see return on their investment. Even basic research done by companies has to be product driven. Issues of interest: why do some cattle get BRD, others dont; animal welfare impact of BRD; BRD/vaccination in beef calves < 4 months old; concern re lack of scientist to do basic research on BRD in the future; interaction of animal behavior/animal husbandry/animal health; better ways to diagnose BRD, especially chute-side. Had short discussion with feedlot practitioner (Dr. Wade Taylor) who joined us.


Adjourned at 4:45 PM.


SEPTEMBER 20, 2007


Meeting re-convened at 8:35 am with open forum. Seven attendees split about evenly between private practitioners and reps from industry (in addition to NC1027 members). Highlights: discussion of BRD in adult dairy cattle. Discussion of pros/cons of SRP vaccines. Comment: would be good if university done in production setting had a technician on site. Lots of good research opportunities are missed. Should also consider sending grad students to busy practices.


Open forum ended at 9:45 pm. Break until 10:00 am.


Business meeting:
Proposal that 2008 meeting be in conjunction with AABP in Charlotte next year (September 24-25) so the NC-1027 will meet September 24-25, 2008.


Bigger question: how do we get more people to come to this meeting? Suggestion: more science would have been interesting (tho it was noted that meeting used to be mostly scientific discussion, and attendance waned in face of that). Proposal by L. Corbeil: also could have a symposium at CRWAD. Ask Bob Ellis to give us half day in Resp section for BRD. Could say sponsored by NC1027wouldnt cost anything. Could build up critical mass for scientific discussion again there.


Comment by R. Fulton: meeting at AABP might be more fruitful (in terms of better interaction with practitioners and improving our ability to gain support from stakeholders) if we get on the AABP program. Could have a session on BRD sponsored by NC1027.


Other discussion: Debate about how much money was the problem with decreasing activity in NC1027. Point made that CSREES cant expect much activity by group when many stations dont get any funding. Consider using electronic means (listserve +/- website) for interaction between group, annual station report submission; and no longer having a formal business meeting. Instead have symposia/presentations involving NC1027 in conjunction with both CRWAD and AABP.


Next Meeting Information: Christopher Chase will organize meeting for next year (September 24-25, 2008). Will ask L. Corbeil to organize half day NC1027-sponsored CRWAD symposium for 2008, Dan Grooms to organize half day NC1027-sponsored AABP symposium in 2008 [they have both since agreed to work on this]. Plan to have business meeting interaction (station report submission) electronically.


Adjourned 10:45 AM.

Accomplishments

Objective 1: To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents. <br /> <br /> <br /> South Dakota reported that respiratory pathogens were similar to preceding years with Pasteurella multicida, Mannheimia haemolytica, and noncytopathic bovine viral virus diarrhea (BVDV) most frequently isolated. Georgia, Iowa and Michigan, with Iowa as the lead institution, submitted a proposal to USDA NRICGP to evaluate the prevalence of M. bovis shedding in calves in cow-calf operations in Iowa, Georgia, and Michigan, and to determine whether shedding of M. bovis at the cow-calf operation predicted occurrence of CPPS in the same calves when they are subsequently placed in feedlots in Iowa. Georgia reported a sensitive and accurate method of identification of BRSV nasal shedding, calves were challenged with virulent BRSV by aerosol. Preliminary information suggests that the qRT-PCR was more sensitive than either VI or FA in identifying infected calves. Results of viral shedding will also be compared to results of BRSV IHC of lung tissue and qRT-PCR of RNA extracted from lung tissue to evaluate agreement between each method of identifying virus in nasal swabs and lung. Michigan reported on additional progress on the porous silicon (PS) - based DNA sensor for the detection of the BVDV and compared to the conductometric platform previously developed. A US patent for the direct electropolymerization used in this application has been applied for. The very rough surface of the nano PS layer prepared the conditions for strong adsorption of polypyrrole (PPy) film on the PS layer without metallic an under layer. Because PS-based platform developed in this work was label-free and metal-free, it is easier to fabricate compared with a metal thin-film electrode and does not need any synthesis procedure for molecular labeling. This amperometric sensitivity detection is more reproducible than conventional optoelectronic or potentiometric methods. Generally, PPy film carries one positive charge for every three monomers. If a receptor molecule specific to BVDV has negative surface charge, there will be an electrostatic attraction between the receptor and PPy film. Even though PPy is intrinsically conductive, the conductivity will be varied with the amount of BVDV coupled with the immobilized receptor molecules. Work on further understanding and refining the conductometric biosensor continues. To date, BVDV positive samples show lower resistance than the negative samples. Optimization of antibody concentrations used in the biosensor has greatly reduced variation, but not to an acceptable level. High variability in sensing signal seems to be the most serious problem in this work. To resolve this problem, a screen-printing system was built. Screen-printed electrodes (SPEs) have the same size and thickness and the same annealing procedure will be used for drying the SPEs. NADC performed a large incidence study. 378 head of beef feeder calves were studied in 4 separate trials over a 30 day period at 3 commercial feedlots in southwestern Iowa. The calves were sourced from consignors located in the states of Georgia, Iowa, and Missouri. All nine serotypes of adenovirus were found to actively infected calves during the 30 days. All calves suffered active infections, many suffered infections with multiple serotypes, and infection with some serotypes was correlated with M. haemolytica infection. Adenovirus may be causing considerable losses to the U.S. beef industry and warrant a commercially-available vaccine. Nasal swabs were analyzed by culture for the presence of P. multocida and M. haemolytica. The former was recovered from a high percentage of calves on arrival at the feedyard (approximately 40%) and became more prevalent during the approximately 30 day study period. M. haemolytica recovery was variable, from 10% to nearly 70% of each group indicating a moderate to high level of stress and/or concurrent viral infection. Shedding of Mannheimia haemolytica was associated with seroconversion to adenovirus. Oklahoma repoted on the prevalence of bovine viral diarrhea virus (BVDV) persistently infected (PI) cattle in beef breeding herds in 30 herds with 4530 calves. The samples collected by ear notches were tested for BVDV antigen using immunohistochemistry (IHC) and antigen capture ELISA (ACE). Animals with initial positives on both IHC and ACE were sampled again for both tests, and serums were collected for viral propagation and sequencing of a viral genomic region, 5-untranslated region (5-UTR) for viral subtyping. Samples were also collected from the dams of PI calves. There were 25 PI calves from 4530 samples (0.55%) and these PI calves were from 5/30 herds (16.7%). Two herds had multiple PI calves and three herds had only one PI calf. Only 1/25 PI calves dam was PI (4.0%). The subtype of all the PI isolates was BVDV1b. Histories of the ranches indicated 23/30 had herd additions of untested breeding females. Twenty-four of 30 herds had adult cowherd vaccinations against BVDV using primarily killed BVDV vaccines at pregnancy examination. Oklahoma reported on BVDV incidence in a cow study. The prevalence of bovine viral diarrhea virus (BVDV) persistently infected (PI) cattle in beef breeding herds was determined in 30 herds with 4530 calves. The samples collected by ear notches were tested for BVDV antigen using immunohistochemistry (IHC) and antigen capture ELISA (ACE). Animals with initial positives on both IHC and ACE were sampled again for both tests, and serums were collected for viral propagation and sequencing of a viral genomic region, 5-untranslated region (5-UTR) for viral subtyping. Samples were also collected from the dams of PI calves. There were 25 PI calves from 4530 samples (0.55%) and these PI calves were from 5/30 herds (16.7%). Two herds had multiple PI calves and three herds had only one PI calf. Only 1/25 PI calves dam was PI (4.0%). The subtype of all the PI isolates was BVDV1b. Histories of the ranches indicated 23/30 had herd additions of untested breeding females. Oklahoma also reported on testing for BVDV in the feedlot where they tested 3023 cattle with 10 cattle determined to be PI (0.33%) using the IHC test on fixed ear notch samples. These include cattle in 32 pens with 7 pens with PI cattle. Two pens have more than one PI calf. Oklahoma also reported on the levels of BVDV virus from PI animals over an 11-month interval. These levels of infectious virus in nasal swabs demonstrate and confirm the high potential of spread of BVDV from PI cattle to susceptible cattle. They monitored viral infectivity and used various tests for PI calves, including the antigen capture ELISA (ACE) on fresh notches and serums, IHC on formalin notches, viral titration (VT) on serum and nasal swabs (NS), and PCR of nasal swab materials, serums, and ear notches. Virus was detected in nasal swabs and serums of PI cattle throughout the study. PI calves do not change test status for ACE and IHC using ear notches and remain positive over time. Selected PI calves were inconsistent for PCR detection of viral genomic material in fresh ear notches. The ACE and IHC remain positive in all of the calves tested from day 0 and at monthly intervals through eleven months of continual testing. Oklahoma also reported on a bovine herpesvirus 1 study. They have obtained a group of almost 50 BHV-1 isolates including current MLV strains, and reference strains such as Cooper (respiratory 1.1 strain) and K-22 (genital 1.2 strain) from respiratory cases in feedlots, genital infections in dairy cattle, neonatal calf disease, and aborted fetuses from heifers/cows recently receiving MLV vaccines. They are currently performing genomic studies to detect differences among the strains. To date several have been examined by restriction endonuclease enzyme fragment polymorphism (RFP). RFP testing using traditional enzymes to date has detected differences among selected field strains and vaccines. Wisconsin reported their BVDV testing rate. BVDV positive testing rate of 0.17% in serum (8 of 4,678), 3.3% in buffy coat (20 of 601), 0.3% in ear notch ELISA (48 of 14,287) and 2.3% in PCR ear notch pools (40 of 1,726).<br /> <br /> <br /> Objective 2: To investigate the basic biology, molecular pathogenesis, and immunopathogenesis of polymicrobial infections including important viral and bacterial agents. <br /> <br /> <br /> South Dakota reported that BVDV PI microarray expression profiling study. Lung leukocytes from PI and non-PI calves were used to determine differences in gene expression profile due to BVDV infection. upregulated includes pro-apoptotic genes like p53 were upregulated while, anti-apoptotic genes and pro-inflammatory genes were down-regulated. California reported on a vaccine trial with a formalin killed alum adjuvanted vaccine and with a H. somni bacterin with a challenge with combined or single pathogens. The BRSV vaccine did not shift the immune response to a Th2 cytokine mediated response and the induction of IgG2 by the bacterin resulted in solid protection from H. somni infection. Cytokine analysis using TaqMan RT-PCR revealed a mixed profile with neither Th1 nor Th2 dominance. California also reported on in vitro development of primary ciliated bovine bronchial epithelial (BE) cell line for use in co-infection studies. Additional studies with bovine turbinate cells to evaluate binding of H. somni to infected and uninfected cells in the presence and absence of inhibitors for binding (such as dextran sulfate and heparin sulfate) are ongoing, including examining the evaluation of BT and BE cells for their ability to alter H.somni binding after various times of infection. California also reported on a study with calves were either immunomodulated with BCG vaccination or mock vaccination potentially to down-regulate Th2 responses, then dually infected with BRSV and H. somni. Necropsy results, H. somni isolation, and clinical signs have been evaluated. Other parameters such as interferon gamma production by PBMC and bronchial lymph node cells are still being evaluated. Georgia establish long term cultures of primary bovine bronchial respiratory epithelial cells (BREC) from samples collected from bronchial brushings of living calves. In preliminary studies, the cells have grown well from bronchial brushings, establishing a differentiated monolayer. Preliminary studies indicate that the cells can be infected with BRSV. Georgia also reported on a leukocyte reactivation from neonatal calves. Proliferative responses against bovine viral diarrhea virus (BVDV) and mycobacterial purified protein derivatives (PPDs) were evaluated. Prior to parturition dams of the calves received a vaccine containing inactivated BVDV, but were not vaccinated against mycobacterial antigens. The results showed that all calves had essentially no IgG in circulation at birth, but comparable and significant levels by day 1. Calves receiving whole colostrum had enhanced leukocyte responses to BVDV antigen 1 and 2 days after ingestion of colostrum. In contrast, calves receiving frozen colostrum, or cell-free colostrum did not respond to BVDV. These results indicated that transfer of live maternal cells from colostrum to neonatal calves enhanced responses to antigens against which the cow had previously responded (BVDV) in the first days of life, but not to antigens for which the cow was naïve (mycobacterial PPD). Kansas reported on work in collaboration with Nebraska that tested the reactivation property and immune response against a gE cytoplasmic tail truncated and a Us9 acidic domain deleted BHV-1 mutants. Several calf and rabbit studies show that both the mutant viruses replicated efficiently in the nasal and ocular epithelium during primary infection. Upon reactivation from latency, both the mutant viruses were not shed and only the respective rescued wild-type viruses were present in the nasal and ocular shedding. Primary immune responses against both the mutant viruses, based on serum neutralization titers were higher than the entire gE and Us9 ORF-deleted viruses (reported last year). Upon dexamethasone induced reactivation, seroconversion was detected only for the respective rescued virus-infected calves. Mississippi is conducting experimental annotation of the three bovine respiratory disease (BRD) pathogens Mannheimia haemolytica, Histophilus somni, and Pasteurella multocida by proteogenomic mapping to improve their ongoing structural annotations and provide functional annotation. In 2007, proteogenomic maps of Mannheimia haemolytica strain PHL213 and P. multocida strain 3480 were produced. Proteins were isolated from each strain in triplicate and analyzed by multi-dimensional protein identification technology (MuDPIT) using two-dimensional liquid chromatography with electrospray ionization tandem mass spectrometry. The resulting mass spectra were searched against their respective protein databases using SEQUEST (Bioworks 3.2 cluster; ThermoElectron). The lists of peptides identified from the genome sequences were compared with the lists identified from protein databases. NADC conducted insertional mutational studies of P. multocida enzymes involved in sialic acid metabolism. These mutants are significantly attenuated in a mouse septicemia model. Also sialic acid mutants may be useful as live vaccines. They constructed sialic acid uptake mutants of bovine P. multocida. Filamentous hemagglutinin (FHA) of B. pertussis plays a critical role in bacterial adherence to host cells and invasion of respiratory epithelial cells. NADC constructed FHA mutants of bovine P. multocida. NADC also developed for isolating ovine lung dendritic cells. A real-time PCR assay for the nonstructural protein 2 (NS2) gene was devised which showed that BRSV can productively infect neonatal ovine lung dendritic cells and alveolar macrophages. Additional real-time PCR assays were developed for proinflammatory and immunomodulatory ovine cytokines. The data indicate that BRSV induces immunomodulatory cytokines (IL-4 and IL-10) during the early stages of infection (days 3 or 5 PI) which would result in a failure to clear BRSV and leave the host susceptible to secondary bacterial infections. Oklahoma has been working on the development of chimeric Mannheimia haemolytica vaccines containing highly immunogenic epitopes from leukotoxin A (LKT) and the outer membrane protein (OMP), PlpE. The five chimeric proteins used as vaccines differed in the number of copies of the LKT and PlpE epitopes they contained and the presence or absence of GST leader sequences or glycine-serine spacers. Mice were intraperitoneally vaccinated with 25, 50 and 75 ¼g quantities of each chimeric protein. Western blot analysis and enzyme-linked immunosorbent assays (ELISA) demonstrated the immune response to the chimeric proteins was strong and specific to PlpE and LKT. Complement-mediated killing assays established that the sera were bactericidal to M. haemolytica in the presence of complement. MTT assays of the hyper-immune murine sera showed that active LKT is neutralized by anti-leukotoxin antibodies formed after vaccination. While all of the chimeric proteins did induce some level of immune response, SAC88 and SAC89 were most immunogenic in that anti-SAC88 and anti-SAC89 sera exhibited the highest complement meditated killing activity and some of the highest levels of LKT neutralizing antibodies. Wisconsin reported on intracellular pathways that result in bovine leukocyte death following exposure to M. haemolytica LKT. LKT internalization can occur by both clathrin-dependent coated pits and lipid rafts. Both of these pathways are inhibited by reducing levels of the scission protein dynamin-3, using siRNA. LKT internalization required actin filament formation, and involves relocation of dynamin from mitochondria to the cell membrane. LKT can be visualized as bound to the outer mitochondrial membrane suing scanning electron microscopy. LKT binding to mitochondria is associated with a drop in mitochondrial membrane potential. These results were obtained using LKT from which LPS was removed, were not observed using LKT produced by an lktC mutant (provided by Dr. S. Highlander, Baylor), and were blocked by addition of an anti-LKT Mab (provided by Dr. Srikumaran, Washington State). The above results were obtained using the BL3 bovine lymphoblastoid cell line, but are being confirmed using peripheral blood neutrophils. A student in the lab is designing an LKT construct to track internalization and transport of LKT in leukocytes. This effort has been aided by Dr. Maheswaran at Minnesota. Wisconsin also reported on the synergyistic effect of active infection with BHV-1 or other bovine respiratory viruses causes more severe pneumonia with M. haemolytica. Infection of primary bovine bronchiolar epithelial cell lines with BHV-1 increases the adhesion of M. haemolytica to these cells. BHV-1 infection increases expression of IL-8, and several inflammatory cytokines. Wisconsin also investigated M. haemolytica outer membrane proteins for their participation in attachment of M. haemolytica cells to bovine bronchial epithelial cells. Recent evidence suggests that one or more lipoproteins are involved in this process. Wisconsin has been assisted in this effort by sera and reagents obtained from Dr. Confer at Oklahoma. Wisconsin also been comparing the responses of bovine pulmonary epithelial and endothelial cells to LPS and LKT. Endothelial cells are more responsive to LPS, exhibiting changes in transepithelial electrical resistance (TEER), expression, of inflammatory cytokines, and morphologic and biochemical changes consistent with apoptosis. Epithelial cells also produced inflammatory cytokines in response to LPS, but did not demonstrate decreased TEER or cytotoxic changes, unless activated neutrophils were included in the incubation mixture. Both epithelial and endothelial cells gave similar signals for TLR-4, suggesting that other differences in signaling are responsible for the dichotomy in response to LPS between the two cell types. Wisconsin has also continued their studies of mechanisms by which Haemophilus somnus causes vasculitis. Using two different isolates of H. somni, that differed in their expression of phosphorylcholine (ChoP) on their LOS, we found that ChoP+ H. somni induced aggregation, while ChoP- H. somni did not. This finding suggested that ChoP on H. somni may interact with platelets, possibly via the PAF receptor, to induce aggregation. However, using an inhibitor of downstream PAF receptor signaling, we found no reduction in platelet aggregation. They inferred that if ChoP binds to the PAF receptor, this event does not induce signaling through this receptor. This observation raises the possibility that platelet aggregation induced by H. somni results from platelet cross-linking of the bacteria via ChoP, rather than initiation of a conventional platelet aggregation pathway. Wisconsin also looked at the effect of H. somni on bovine platelets and bovine pulmonary artery endothelial cells. Endothelial cells were incubated with platelets that had been activated with ADP, H. somni or H. somni LOS. Incubation with activated bovine platelets significantly increased expression of adhesion molecules (ICAM-1, E-selectin) and tissue factor, as measured by flow cytometry, real-time PCR, and western blot analysis. Activated platelets also up-regulated expression of endothelial cell IL-1², MCP-1, and MIP-1± as determined by real-time PCR and an IL-1² ELISA. Bovine platelets, but not H. somni, were internalized by bovine endothelial cells as visualized by transmission electron microscopy. This observation raises the possibility that endocytosis of bovine platelets may contribute, in part, to endothelial cell activation and cell stress. These findings suggest that H. somni activated bovine platelets might play a role in promoting vasculitis by increasing endothelial cell pro-inflammatory responses. The above studies involved collaboration with Dr. Corbeil (UC-San Diego and UC-Davis) and Dr. Inzana (Virg. Tech). Wisconsin also looked at the most severe manifestation of H. somnus infection, thrombomeningoencephalitis (TME). Cerebral microvascular endothelial cells are the primary cellular constituent of the blood brain barrier, which protects the central nervous system from inflammation and infectious agents. We developed an in vitro model and found that binding of H. somnus to bovine brain endothelial cells (BBEC) can be inhibited by high concentrations of heparin, or by pretreating the BBECs with heparinase. Addition of hyaluronic acid, a nonsulfated glycosaminoglycan, had no effect. These data suggest a role for sulfated glycosaminoglycans in the adhesion of H. somnus to BBECs. BBECs up-regulated TNF-±, IL-1², tissue factor, and ICAM-1 expression, following exposure to H. somnus. They also down-regulated their expression of occludin, a tight junction protein, and exhibited a decreased ability to activate protein C. Increased permeability of BBECs in response to viable, but not killed H. somnus, was observed as measured by decreased transepithelial electrical resistance (TEER) and by albumin leakage across the monolayers. These changes in monolayer permeability could be blocked by addition of a myosin light chain kinase inhibitor. The observed alterations in the BBECs could play a role in the development of vasculitis, thrombosis and inflammation that results in blood brain barrier dysfunction and bacterial invasion of the brain parenchyma. These studies involved collaboration with Dr. Corbeil and Dr. K.S. Kim (Johns Hopkins). <br /> <br /> <br /> Objective 3:To develop management and prevention strategies that incorporate new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss. <br /> <br /> <br /> South Dakota reported on the effect of antigen trafficking following vaccination. Vaccinated with a NCP BVDV vaccine had longer antigen retention than either of the CP BVDV vaccines. Georgia has been working on developing siRNA molecules to inhibit replication of BRSV. Initials results have confirming inhibition of viral replication in vitro. Iowa was the lead station on a collaboration with GA, NADC and IA stations, and included the study of beef calves from GA farms that were sampled on arrival at a transit receiving station in GA, then sampled again at 1 and 2 months after arrival at IA feedlots. A total of 11 farms in GA were sampled. Sampling could detect 100% of calves as positive (one instance). A farm was arbitrarily considered as focally infected if < 10% of calves were infected, and widely infected if the percentage was higher. Only 2 of 11 GA farms were widely infected. Calves from those farms stayed infected at the same or higher proportion upon commingling at feedlot. Calves from the 9 naïve or focally infected farms all became widely infected upon commingling at feedlot. All calves from the second shipment were negative at farm. Overall, the pattern was a shift from naïve or focally infected to widely infected at feedlot. Iowa also tested several antimicrobials with potential usefulness against North American strains of M. bovis. Forty two isolates of M. bovis recovered from Midwest outbreaks in 1999 to 2006 were tested. Of these, 7 were from BRD cases and the rest from mastitis cases, reflecting the shift in case-loads handled by the Iowa Veterinary Diagnostic Lab. Two antimicrobials, ceftiofur and tilmicosin, were previously shown to be ineffective in-vitro against M. bovis in the previous study, and this was confirmed in this study. Enrofloxacin and florfenicol were effective against a majority (over 90%) of strains. Tulathromycin was shown to be ineffective against all strain tested. Michigan has initiated a regional BVDV control program to control and eradicate BVDV. The purpose is to identify benefits and obstacles of such a program and demonstrate a feasible model that may be adopted by other parts of the US. Wisconsin conducted an investigation of the relationships between air quality, a variety of environmental risk factors, and calf respiratory health were studied in 13 naturally ventilated calf barns during winter. A minimum of 12 pre-weaned calves were randomly selected and scored for the presence of respiratory disease in each barn. An air sampling device was used to determine airborne bacteria colony forming units per cubic meter (cfu/m3) of air in calf pens and central alleys within the barns. Temperature and relative humidity were recorded in each calf pen, the barn alley, and outside the barn. Samples of bedding were collected in each pen and dry matter was measured. Pen bedding type and a calf nesting score (degree to which the calves could nestle into the bedding) was assigned to each barn. Calf numbers, barn and pen dimensions, ridge, eave, and curtain openings, and exterior wind speed and direction were determined and used to estimate building ventilation rates. Factors that were significantly associated with a reduction of prevalence of respiratory disease were reduced pen log10 cfu/m3 on BAP, presence of a solid barrier between each calf pen, and increased ability to nest. Individual calf pen log10 cfu/m3 were significantly different from barn alley log10 cfu/m3 on both BAP and EMB. Significant factors associated with reduced calf pen log10 cfu/m3 on BAP were increasing pen area, increasing number of open planes of the calf pen, decreasing pen temperature, and wood-particle bedding. Significant factors associated with reduced alley log10 cfu/m3 on BAP were increased ventilation changes per hour, increased barn volume per kg of calf, reduced pen log10 cfu/m3, and barn type.<br /> <br /> <br /> <br />

Publications

Journal Publications:<br /> <br /> <br /> 1. Ridpath JF, CS Mark, CCL Chase, AC Ridpath, and JD Neill. Febrile response and decrease in circulating lymphocytes following acute infection of white tail deer fawns with either a BVDV1 or a BVDV2 strain. J Wildlife Dis. in press.<br /> <br /> <br /> 2. Zimmerman AD, Buterbaugh RE, Herbert JM, Hass JM, Frank NE, Luempert III LG, Chase CCL. Efficacy of bovine herpesvirus-1 inactivated vaccine against abortion and stillbirth in pregnant heifers. JAVMA, 2007; 231:13861389. <br /> <br /> <br /> 3. Corbeil LB, Arnold KF, Kimball R, Berghaus L, Gershwin LJ. Specificity of IgG and IgE antibody responses to Haemophilus somnus infection of calves. Vet Immunol Immunopathol. 2006 Sep 15;113(1-2):191-9. <br /> <br /> <br /> 4. Berghaus LJ, Corbeil LB, Berghaus RD, Kalina WV, Kimball RA, Gershwin LJ. Effects of dual vaccination for bovine respiratory syncytial virus and Haemophilus somnus on immune responses. Vaccine. 2006 Aug 14;24(33-34):6018-27. <br /> <br /> <br /> 5. Donovan DC, Reber AJ, Gabbard JD, Aceves-Avila M, Galland KL, Holbert KA, Ely LO, Hurley DJ. Effect of maternal cells transferred with colostrum on cellular responses to pathogen antigens in neonatal calves. Am J Vet Res, 2007; 668:778-782.<br /> <br /> <br /> 6. Wiggins MC, Woolums AR, Sanchez S, Hurley DJ, Cole DJ, Ensley DT, Pence ME. Prevalence of Mycoplasma bovis in backgrounding and stocker cattle operations. J Am Vet Med Assoc. 2007; 230:1514-1518. <br /> <br /> <br /> 7. Al-Mubarak-A, Simon, J, Coats, C, Okemba, JD, Burton, MD, and Chowdhury, SI. Glycoprotein E(gE) specified by Bovine herpesvirus type5 (BHV-5) enables trans-neuronal virus spread and neurovirulence without being a structural component of enveloped virions. Virology. 2007; 365:398-409.<br /> <br /> <br /> 8. Butchi, NB, Jones, C, Perez, S, Doster, A, and Chowdhury,SI. Role of Envelope protein Us9 in the anterograde transport of BHV-1 following reactivation in the Trigeminal Ganglia . J. Neurovirology. 2007; 13(4):384-388.<br /> <br /> <br /> 9. Muhammad-Tahir, Z, Alocilja, EC, and Grooms, DL. Indium Tin Oxide-Polyaniline Biosensor: Fabrication and Characterization. Sensors. 2007; 7:1123-1140.<br /> <br /> <br /> 10. Lee, S-R, Pharr, GT, Boyd, BL, Pinchuk, LM*. Bovine viral diarrhea viruses modulate toll-likereceptors, cytokines and co-stimulatory molecules genes expression in bovine peripheral blood monocytes.Comparat Immunol Microbiol Infect Dis. 2007; doi:10.1016/j.cimid.2007.06.006S.<br /> <br /> <br /> 11. Thomas, CJ, Hoet, AE, Sreevatsan, S, Wittum, TE, Briggs, RE, Duff, GC, and Saif, LJ. Transmission of bovine coronavirus and serologic responses in feedlot calves under field conditions. Am J Vet Res. 2006; 67:1412-20. <br /> <br /> <br /> 12. Fach, SJ, Brockmeier, SL, Hobbs, LA, Lehmkuhl, HD, and Sacco, RE. Pulmonary dendritic cells isolated from neonatal and adult ovine lung tissue. Vet Immunol Immunopathol. 2006; 112:171-82.<br /> <br /> <br /> 13. Kawashima, K, Meyerholz, DK, Gallup, JM, Grubor, B, Lazic, T, Lehmkuhl, HD, and Ackermann MR. Differential expression of ovine innate immune genes by preterm and neonatal lung epithelia infected with respiratory syncytial virus. Viral Immunol. 2006; 19:316-23.<br /> <br /> <br /> 14. Fach, SJ, Meyerholz, DK, Gallup, JM, Ackermann, MR, Lehmkuhl, HD, and Sacco, RE. Neonatal ovine pulmonary dendritic cells support bovine respiratory syncytial virus replication with enhanced interleukin IL-4 and IL-10 gene transcripts. Viral Immunol. 2007; 20:119-30. <br /> <br /> <br /> 15. Meyerholz, DK, Gallup, JM, Lazic, T, de Macedo, MM, Lehmkuhl, HD, and Ackermann MR. Pretreatment with recombinant human vascular endothelial growth factor reduces virus replication and inflammation in a perinatal lamb model of respiratory syncytial virus infection. Viral Immunol. 2007; 20:188-96. <br /> <br /> <br /> 16. Gatto, NT, Estes, DM, Confer, AW, Whitworth, LC, and Murphy, GL. Lung Lesions in SCID-bo and SCID-bg Mice after Intratracheal inoculation with Wild-type or Leucotoxin-deficient Mutant Strains of Mannheimia haemolytica Serotype 1. J Comp Patho. 2006; 134:355-365.<br /> <br /> <br /> 17. Atapattu, DN and Czuprynski CJ. Mannheimia haemolytica leukotoxin binds to lipid rafts in bovine lymphoblastoid cells (BL-3) and is internalized in a dynamin-2 and clathrin-dependent manner. Infect Immun. 2007 Aug 6; [Epub ahead of print] <br /> <br /> <br /> 18. Behling-Kelly, E, Vonderheid, H, Kim, KS, Corbeil, LB, and Czuprynski, CJ. Roles of cellular activation and sulfated glycans in Haemophilus somnus adherence to bovine brain microvascular endothelial cells. Infect. Immun. 2006; 74:5311-5318. <br /> <br /> <br /> 19. Behling-Kelly, E, McClenahan, D, Kim, KS, and Czuprynski, CJ. Viable Haemophilus somnus induce myosin light chain kinase-dependent decrease in brain endothelial cell monolayer resistance. Infect Immun. 2007; Jun 25; [Epub ahead of print] <br /> <br /> <br /> 20. Kuckleburg, CJ, Elswaifi, SF, Inzana, TJ, and Czuprynski, CJ. Expression of phosphorylcholine by Histophilus somni induces bovine platelet aggregation. Infect Immun. 2007; 75:1045-1049. <br /> <br /> <br /> 21. Kuckleburg, CJ, McClenahan, DJ, and Czuprynski, CJ. Platelet activation by Histophilus somni and its lipooligosacharride induces endothelial cell proinflammatory responses and platelet internalization. Shock. 2007, Jul 19; [Epub ahead of print] <br /> <br /> <br /> 22. Lago, A, McGuirk, SM, Bennett, TB, Cook, NB, and Nordlund, KV. Calf respiratory disease and pen microenvironments in naturally ventilated calf barns in winter. J. Dairy Sci. 2006; 89:4014-4025. <br /> <br /> <br /> 23. Sylte, M, Inzana, T, and Czuprynski, CJ. Role of tumor necrosis factor in endothelial cell apoptosis caused by Haemophilus somnus lipooligosaccharide. Vet. Immunol. Immunopathol. 2006; 110:303-309.<br /> <br /> <br /> <br /> Abstracts and Conference Proceedings: <br /> <br /> <br /> 1. Ridpath JF, CS Mark, CCL Chase and JD Neill. Experimental acute BVDV infection in white tail deer fawn. Proceedings of the 39th annual conference of the American Association of Bovine Practitioners, Saint Paul, MN, September 21-23, 2006, p. 271.<br /> <br /> <br /> 2. Ridpath JF, BE Hessman, JD Neill, RW Fulton, DL Step, A Zimmerman and CCL Chase. Evaluating stability, size requirements, viral load and pooling of ear notch samples in BVDV testing. Proceedings of the 49th annual conference of the American Association of Veterinary Laboratory Diagnosticians, Minneapolis, MN, October 12-18, 2006, p. 213.<br /> <br /> <br /> 3. Tigabu B, A Rosa, G Rosa and CCL Chase. Comparison of immune gene expression in normal and BVDV persistently infected cattle. Abstract 2. 66th annual meeting of the North Central Branch of the American Society of Microbiology, Brookings, SD, October 20-21, 2006, p. 21. <br /> <br /> <br /> 4. Halaweish S, Y Fang, I Halaweish, L Braun and C Chase. Expression of recombinant BVDV-nonstructural proteins to study the humoral immune response against BVDV in cattle. Abstract 1. 66th annual meeting of the North Central Branch of the American Society of Microbiology, Brookings, SD, October 20-21, 2006, p. 21. <br /> <br /> <br /> 5. Tigabu B, A. Rosa, G. Rosa, CCL Chase. Comparison of Immune Gene Expression in Normal and BVDV Persistently Infected Cattle. Abstract, p73. 87th annual meeting of Conference of Research Workers in Animal Disease, Chicago, IL, December 3-5, 2006.<br /> <br /> <br /> 6. Halaweish S, Y Fang, I Halaweish, L Braun, C Chase. Expression of recombinant BVDV-Nonstructural Proteins to Study the Humoral Immune Response against BVDV in Cattle. Abstract, p73. 87th annual meeting of Conference of Research Workers in Animal Disease, Chicago, IL, December 3-5, 2006.<br /> <br /> <br /> 7. Mahmoud MM, LJ Braun, A Hippen, M Thomas, and CC Chase. Effect of source of selenium and vitamin E supplementation on performance and immune response in young dairy calves. Abstract, p84. 87th annual meeting of Conference of Research Workers in Animal Disease, Chicago, IL, December 3-5, 2006.<br /> <br /> <br /> 8. Rosenbusch, R. F. Antimicrobial susceptibility of recent isolates of Mycoplasma bovis. Academy of Veterinary Consultants, Spring 2007 Meeting, Oklahoma City, OK, 2007.<br /> <br /> <br /> 9. S-R Lee1*, G. T. Pharr1, L. M. Pinchuk1. Bovine Viral Diarrhea Viruses Modulate Type I IFN and Pro-Inflammatory Cytokines Production via Toll-Like Receptor-Dependent Mechanisms. S.-R. Lee1*, G. T. Pharr1, L. M. Pinchuk1. Oral presentation at CRWAD, Chicago, IL, Dec.3-5, 2006.<br /> <br /> <br /> 10. Lawrence, ML, SC Burgess, S Bridges, B Nanduri, N Wang, and F McCarthy. Experimental annotation of bovine respiratory disease pathogen genomes by proteogenomic mapping. Plant and Animal Genome XV Conference, San Diego, California, 2007.<br /> <br /> <br /> 11. Confer A. Pathogenesis of pulmonary infections with Pasteurellaceae. Proceedings of the 24th Symposium of the Veterinary Comparative Respiratory Society. Jena, Germany, 2006.<br /> <br /> <br /> 12. Trojan B, Ayalew S, Montelongo M, Confer AW. Intranasal Vaccination of Dairy Calves with Mannheimia haemolytica Chimeric PlpE-LKT (SAC89). ASM abstract for presentation at the annual meeting in Toronto, Canada, May 2007.<br /> <br /> <br /> 13. Ayalew S, Stevens KD, Montelongo M, and Confer AW. Mannheimia haemolytica Chimeric Protein Vaccine Composed of the Major Surface-Exposed Epitope of Outer Membrane Lipoprotein PlpE and the Neutralizing Epitope of Leukotoxin. ASM abstract for presentation at the 107th annual meeting in Toronto, Canada, May 2007.<br /> <br /> <br /> 14. Fulton, RW, Braziel, BC, Kautz, K, Burge, LJ, Weaver, GD, and Hessman, B. Bovine Respiratory Syncytial Virus, Bovine Coronavirus and Bovine Viral Diarrhea Virus Diagnosis by PCR Testing of Nasal Swabs. American Association of Bovine Practitioners Annual Meeting Proceedings. St. Paul, MN, September 21-23, 2006. <br /> <br /> 15. Ridpath, JF, Hessman, BJ, Neill, J, Fulton, RW, and Step, DL. Parameters of Use of Ear Notch Samples for BVDV Testing: Stability, Size Requirements and Viral Load. American Association of Bovine Practitioners Annual Meeting Proceedings. St. Paul, MN., September 21-23, 2006. <br /> <br /> <br /> 16. Fulton, RW, Whitley, EM, Johnson, BJ, Kapil, S, Ridpath, JF, Burge, LJ, Cook, BJ, and Confer, AW. Bovine Viral Diarrhea Virus Persistent Infectious in Beef Breeding Herds; Utilization of Immunohistochemistry and Antigen Capture ELISA on Ear Notches. 49th Annual Meeting of AAVLD. Minneapolis, MN, October 12-15, 2006. <br /> <br /> <br /> 17. Fulton, RW, Elam, N, Hessman, BE, Johnson, BJ, Kapil, S, Ridpath, JF, Burge, LJ, Braziel, BC, Kautz, K, and Reck, A. Bovine Viral Diarrhea Virus Persistently Infected and Acutely Infected Calves: Assays for Viral Infectivity, Polymerase Chain Reaction Analysis, and Antigen Detection. 49th Annual Meeting of AAVLD. Minneapolis, MN, October 12-15, 2006. <br /> <br /> <br /> 18. Fulton, RW. Detecting Persistently Infected BVDV Cattle in Cow-Calf Herds. Academy of Veterinary Consultants Meeting, Denver, CO, December 1, 2006.<br /> <br /> <br /> Other (Ph.D Dissertations): <br /> <br /> <br /> 1. Tigabu B., PhD. The Effect of Bovine Viral Diarrhea Virus on Immune Gene Expression of Bovine Leukocytes. South Dakota State University, 2007.<br /> <br /> <br /> 2. Chris Kuckleburg, PhD. Bovine platelets activated by Histophilus somni induce endothelial cell activation and apoptosis. University of Wisconsin-Madison, 2007. <br /> <br /> <br /> 3. Erica Behling-Kelly, PhD. Interactions Between Haemophilus somnus and Bovine Brain Endothelial Cells. University of Wisconsin-Madison, 2007.<br /> <br /> <br /> <br />

Impact Statements

1. Objective 1: Continued monitoring will help track new and emerging diseases. The finding that qRT-PCR may be a more sensitive method to identify shedding of BRSV than traditional methods will allow researchers, diagnostic laboratories, and clinicians in the field better identify this common cause of respiratory disease and thus implement preventative strategies to reduce the cost of morbidity and mortality associated with BRD.
2. Objective 1 (continued): A rapid detection of BVDV in cell culture media using a biosensor was demonstrated. Detection of cattle persistently infected with BVDV using ear notches was shown to be most consistent when compared to serum and nasal swabs. Further development may make this a useful tool for the rapid and economical identification of BVDV PI cattle. The high incidence of adenovirus infection in calves from multiple sources together with correlation with M. haemolytica shedding indicate the possibility that adenovirus may be causing substantial economic loss in feedlots.
3. Objective 1 (continued): Field studies with naturally occurring disease will investigate multiple viral etiologies interacting with M. haemolytica and P. multocida. Also these field studies will permit evaluation of current viral and bacterial vaccines along with newly developed vaccines. The diversity (antigenic) of BVDV will be further examined to determine appropriateness/relevance of current and future BVDV vaccines to control BVDV.
4. Objective 2: Understanding virus cell interactions will provide new targets for intervention. When the experiments are complete they will provide information on the mechanism(s) involved in the disease synergy that occurs when calves are dually infected with BRSV and H. somni.
5. Objective 2 (continued): The research showing that colostrum containing maternal cells improved the immune response of calves in the first days of life will help producers by proving that, if calves in their herd are developing infections early in life, cell containing (fresh) colostrum is appropriate to administer. This will help producers justify the increased effort to make certain calves get fresh colostrum as much as possible, rather than frozen colostrums.
6. Objective 2 (continued): The mutant BHV-1 viruses maybe a better vaccine virus than the current gE-deleted marker vaccine since it will induce a better primary immune response relative to gE-deleted virus yet like the gE-deleted virus it will be safe because the vaccine virus will not be shed following reactivation from latency.
7. Objective 2 (continued): Improved annotation of the Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni genomes will enable functional genomics investigations and systems modeling for these important bovine respiratory disease pathogens.
8. Objective 2 (continued): The results of the study on the differential expression of proteins related to immune responses such as cell adhesion, apoptosis, antigen uptake, processing and presentation, acute phase response proteins, MHC class I- and II- related proteins support our hypothesis that BVDV may compromise anti-viral immune responses by altering the expression of the molecules involved in immune function of professional antigen presenting cells.
9. Objective 2 (continued): The ecology of P. multocida mucosal colonization and disease pathogenesis is very poorly understood in any species. The finding that filamentous hemagglutinin and LPS/LOS decoration with sialic acid are both important features for mucosal infection in turkeys may lead to new methods of disease control in this and other species. The identification, cloning, and production of subunit components of M. haemolytica and P. multocida offer opportunity for new bacterial vaccines to control BRD.
10. Objective 3 : The impact of the siRNA research is that, if siRNA is proven to prevent disease following BRSV infection of cattle, siRNA can be administered to cattle in the field to prevent disease due to BRSV infection. While siRNA is still relatively expensive, it is expected that the cost of siRNA will decrease as the technology is further developed. Because siRNA molecules are relatively simple to make and quite specific in the effect on given viruses, the technology could potentially be applied to prevent disease due to a variety of viral infections in cattle.
11. Objective 3 (continued): Data on recovery of M. bovis from the noses of calves will need to be correlated with health status and production parameters in the future. Antimicrobial activity data against M. bovis must be obtained for all drugs in current use in BRD since therapy is the main management tool used for these infections. The Michigan regional BVDV Eradication program is being designed conceptually. Once initiated, the potential benefits and usefulness of such a program will be documented.

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Report Information

Participants

" *Woolums, Amelia (awoolums@vet.uga.edu)- U of Georgia;
" *Czuprynski, Chuck (czuprync@svm.vetmed.wisc.edu)  U of Wisconsin;
" *Ricardo Rosenbusch (rfrosenb@iastate.edu) - Iowa State U;
" *Dan Grooms (groomsd@cvm.msu.edu) - Michigan State U;
" *Fulton, Robert (Robert.Fulton@okstate.edu)  Oklahoma State U;
" *Chase, Chris (christopher_chase@sdstate.edu) - South Dakota State U;
" Dan Thomson (dthomson@vet.k-state.edu) - Kansas State U;
*voting member

Brief Summary of Minutes

AGENDA:


Wednesday September 24;


8:30 am: Welcome;


8:40-10:00 am: Station reports;


10:00-10:15: Break;


10:15 am - 12:00 noon: Station Reports;


12:00 - 1:00 pm: Lunch;


1:00-2:15 PM: Conference call with Gary Sherman and Peter Johnson, CSREES;


2:15-2:45 pm: Discussion of presentation to be made to AABP on the problem of insufficient funding for bovine disease research;


2:45-4:30 pm: Business meeting;


4:30-5:00 pm: Complete station reports;


5:00 pm: Adjourn;



Thursday September 25, 2008;


8:00-9:00 am: Open forum for discussion of bovine respiratory disease.






MINUTES:


September 25, 2008


2:45-4:30 pm: Business meeting.


Meeting began at 2:45 pm. Brief discussion of midterm report being written and organized by Ricardo Rosenbusch. Update on National BRD Symposium (to be sponsored by NC-1027 and held August 5 and 6, 2009) presented by Amelia Woolums, chair of organizing committee (organizing committee: Chris Chase, Robert Fulton, Ricardo Rosenbusch). Discussion of BRD session organized by Lynette Corbeil for NC-1027 at CRWAD, Monday December 7, 2008. Plan made for 2009 meeting of Technical Committee to occur on Friday morning August 7, after the BRD Symposium, at the Crown Plaza in Colorado Springs, CO. Chris Chase agreed to serve as President of Technical Committee for 1 more year, and Amelia Woolums will serve as Secretary of Committee for 1 more year.





Notes from Teleconference with Dr. Gary Sherman and Dr. Peter Johnson:


Committee asked if BRD would remain on the list of fundable diseases for NRI. Dr. Johnson said that he couldnt say for sure, but if BRD is still an important issue, it is likely to stay on list. If stakeholders continue to identify BRD as a problem in routine feedback to USDA CSREES, and particularly at the stakeholder meeting scheduled for 2010, it will be very likely to stay on list. Given that, from year to year things can change regarding priorities for funding.


Chuck Czuprynski noted that multistate projects were mentioned as a specific target for funding in the AES call for proposals this year. Committee asked if this indicated some special attention to multistate projects like NC-1027, or was this in reference to something else? Dr. Sherman said that no significant policy changes had been made on how AES funding is distributed; this is still up to the Expt. Station Directors. If special mention was made most likely just reflected local considerations.


Drs. Johnson and Sherman outlined upcoming reorganization of CSREES, and the timetable for the RFP for FY 2009. Highlights:


NRI is going to become the Agricultural and Food Research Initiative (AFRI) in 2009; this will be announced in Nov. or Dec. 2008. Formation of AFRI is not a cause for concern; in fact, its formation in the new Farm Bill was intended to provide a fresh look at how agricultural research should be done/funded.


Big change: NRI could only fund research projects and integrated (combined research/education/extension) projects. AFRI will be able to fund research, integrated, OR solely education or extension projects. Not confirmed that such projects will be solicited, but they could be.


Other changes: AFRI is authorized for funding up to $700 million, though actual amount funded will depend on appropriation. NRI had only been authorized for funding up to $500 million (though the largest amount every appropriated was $191 million in 2008).


Indirect costs for AFRI funded projects will be capped at 22% (NRI: capped at 20%). AFRI has authority to fund projects for up to 10 years (though this will likely be very rare in practice; NRI had authority to fund projects for up to 5 years).


AFRI will have 6 focus areas, the area relevant to animal disease will be Animal Health, Animal Production, and Animal Products (in NRI, the relevant area was Genomics and Biosecurity).


Things not changed in AFRI vs NRI: Both basic and applied studies will be funded. Matching requirements will be essentially the same. Postdoc awards, etc. will still be funded.


Deadlines for FY 2009: The NRI RFA has not been posted because CSREES has been involved in working toward reorganization. NRI will cease to exist as of this year, thus there will be no RFA for NRI (proposals currently funded under NRI will continue under NRI for now but eventually will be switched to AFRI). All deadlines for proposals to be funded through AFRI will be announced when the description of AFRI and other products of the reorganization of CSREES is announced in Nov or Dec 2008. Dr. Johnson estimates that letters of intent for AFRI will be due in January 2009, and proposals will be due in March 2009. In 2010, AFRI may go back to a fall application deadline, not certain.


Change in application process: Because many have noted that the Pure Edge system is cumbersome, its use by Grants.gov is to be discontinued. Proposals will be submitted using pdf files [note: as of now it wasnt clear exactly how this would work].


Other changes: CSREES is being converted into the National Institute for Food and Agriculture (NIFA). The director of NIFA will be a political appointee (but will not require Congressional approval) with a 4 to 6 year term who reports to the Secretary of Agriculture. This is [at least in part] a result of land grant universities wishing for research, extension, and education activities to be more visible to the Secy. of Ag. The current Head of CSREES, Colleen Heffren, is a civil servant. Not sure what will happen to her position. Drs. Johnson and Sherman anticipate that appointing the Director of NIFA may not be the highest priority for the new President, so they will likely operate under an Interim Director until sometime well into 2009 [or perhaps later]. The actual structure of NIFA will be determined by the Director, so for the time being organization will likely change little, but may change much in the future. So this may impact where Dr. Johnson and Dr. Sherman are placed in the NIFA organization.


NIFA will have a Chief Scientist who will be under the Director and will be equivalent to an Undersecretary. The Chief Scientist will oversee the Directors of the 6 units of NIFA (which will have the same names as the 6 programs in AFRI, which should make it more clear how activities are administered). Thus, there will be a Director of Animal Health, Animal Production, and Animal Products who will be responsible for programs related to animal disease research, education, and extension.


The Chief Scientist will also oversee APHIS and other units formerly included in CSREES.


Significant change: USDA ARS and CSREES will be combined in NIFA. This should improve interaction and communication regarding agricultural research funded by USDA.


Dr. Johnson pointed out that, funding-wise, BRD fares well in NRI. In the past fiscal year 6 research proposals and 2 conference proposals related to BRD [including NC-1027s BRD Symposium proposal for $10,000] were funded at a total of just over $2.2 million. This represented about 20% of the funds available through the Animal Protection program.


The committee asked if the formation of NIFA and AFRI would have any impact on multistate projects. Dr. Sherman said no change was anticipated, though the formula (which directs the funding of projects) could always change.


NIFA wont have a budget until late in 2009 and so until then will operate on a continuing resolution.


Dr. Johnson pointed out that the 406 Integrated Programs were joined with NRI to form AFRI; since the 406 programs were funded at $45 million in 2008, and NRI was funded at $191 million, this means that the total amount of money available through AFRI should be relatively increased over what was available in NRI.


The committee asked about the future of the CAP program. The CAP program has strong support, which suggests that it will probably continue. Within the last year all 3 of the current CAP projects were renewed, suggesting there will be no call for new CAP proposals in 2009. There may be a call for new proposals in 2010.


Important note about the 1433 program: Dr. Sherman pointed out that in each of several recent years, the President included no funding for the 1433 program, and each year Congress returned it (this is one of the programs through which Formula Funds are awarded). However, this year the Senate also included $0 for 1433 (proposed funding was $5 million, typical of recent years). The budget still has to go through the House and the Conference Committee, but the fact that the Senate budgeted nothing for this program does not bode well for the program. Moreover, the National Veterinary Medical Services (NVMS) Program (which provides loan forgiveness for new grads working in underserved areas of vet med) was funded by the Senate at $5 million. Dr. Sherman couldnt say for sure, but this suggests that the Senate had the idea to stop funding the 1433 Program and to use the funds instead for the NVMS Program. Will have to stay tuned to see how each of these programs is ultimately funded.

Next meeting of NC-1027 Technical Committee: Friday August 7, 2009, at the Crown Plaza Hotel in Colorado Springs, CO (the morning after the BRD Symposium).


Members in attendance agreed that we should not plan to do station reports in 2009, but rather use the time to discuss the BRD Symposium and to engage in serious discussion about whether a new NC-1027 project should be submitted when the current project ends, in light of the greatly decreased rate of attendance at the annual meeting in the past 4 years.


September 25, 2008


8:00-9:00 am: Open forum for discussion of bovine respiratory disease.


Approximately 70 people from AABP Convention were in attendance; these attendees posed questions to members of NC-1027, and shared their experience with BRD, leading to good discussion. Dr. Ricardo Rosenbusch took notes and Dr. Dan Grooms has offered to summarize the notes and will ask Dr. Riddell (AABP Exec. Director) if the notes can be published in the AABP newsletter.


Minutes were recorded by Dr. Amelia Woolums.

Accomplishments

ACCOMPLISHMENTS:<br /> <br /> <br /> OBJECTIVE 1: To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents. <br /> <br /> <br /> Kansas evaluated the effect of ear-notch sample handling on the reliability of antigen-capture ELISA testing for BVDV. Neither the volume of PBS added to sample, nor time after collection affected the outcome of the assay. A comparison of pooled testing to individual testing of ear-notch samples was also made. Based on test detection limits and economic factors, a pool size of 5-10 notches appeared to be the most advantageous for detecting BVDV. Kansas also explored the use of cardiac troponin level measurements in blood as an indicator to aid in field diagnosis of bovine respiratory disease. An I-STAT 1 hand-held blood analyzer was used to measure Troponin-I in healthy and BRD-affected calves, and affected calves had significantly higher Troponin I levels in blood. Michigan continued the development of a biosensor chip to be used in the detection of BVDV persistently infected cattle. The rapid conductimetric sensors developed to date still lack the sufficient sensitivity for field use. South Dakota reported on diagnostic findings of bovine respiratory bacteria and viruses for 2008. Comparative use of various diagnostic tests for the detection of persistent BVDV were also reported. Wisconsin reported on bacterial and viral diagnostic results from BRD cases. Oklahoma reported on difficulties using a commercial antigen-capture ELISA test for BVDV persistence. The erratic results could be overcome by freezing ear-notch washes before testing.<br /> <br /> <br /> <br /> <br /> OBJECTIVE 2: To investigate the basic biology, molecular pathogenesis, and immunopathogenesis of polymicrobial infections including important viral and bacterial agents. <br /> <br /> <br /> California examined the in vitro adherence of Histophilus somni on bovine turbinate cells infected with BRSV, and the expression of virulence factors by H. somni under these culture conditions. No significant effect of the viral infection on adherence or expression of virulence factors of H. somni was found, even though concurrent infection of calves with these two agents resulted in increased severity of disease and persistence of H. somni in lung. Georgia reported the successful cultivation of bovine respiratory ciliary epithelial cells. When these cells were infected with BRSV, a transient and elevated mRNA synthesis to the chemokines IL-8 and RANTES, and the adhesion molecule ICAM-1 were detected. Kansas investigated the use of bovine adrenal gland endothelial cell monolayers as a permeability barrier for macromolecules. Substance P and histamine were reported to cause increased permeability across the monolayer, while TNF and LPS did not. Mississippi reported on initial proteogenomic mapping efforts for Mannheimia haemolytica, Pasteurella multocida, and H. somni. Protein profiling was accomplished on bovine monocytes infected with a cytopathic or noncytopathic BVDV virus. Several protein kinases were reduced in expression in monocytes infected with cytopathic BVDV. Nebraska studied the transcriptional activator bICPO of BHV-1. A mutant of the virus with single amino acid changes in the zinc finger domain of b ICPO was shown to grow inefficiently in bovine cells in vitro and calf mucosa in vivo. Other studies focused on the enhancing interaction of host cell transcription factors with viral transcription factors in this model. Studies are on-going on the effect of latency reactivation gene protein of BHV-1. The protein appears to spare neurons from apoptosis induced by exogenous agents. Wisconsin continues to study intracellular pathways that result in leukocyte death following exposure to M. haemolytica leukotoxin. Once internalized, the leukotoxin is transported to the mitochondria. This transport is correlated with actin filament formation and dynamin relocation from mitochondria to the cell membrane. Bovine pulmonary epithelial cells were compared to endothelial cells in their response to LPS. Endothelial cells were shown to be more responsive to LPS, resulting in increased transepithelial resistance, expression of inflammatory cytokines and apoptosis. Epithelial cells only responded with increased inflammatory cytokines. BHV-1 infection of bovine bronchiolar epithelium cell lines resulted in increased adherence of M. haemolytica to these cells. This adherence is mediated by two outer membrane proteins of the bacteria. Wisconsin also continued studies on expression of surface molecules by brain microvascular endothelial cells infected with H. somni.<br /> <br /> <br /> <br /> <br /> OBJECTIVE 3: To develop management and prevention strategies that incorporate new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss. <br /> <br /> <br /> California investigated the use of recombinant GST fusion products of 3 surface immunoglobulin-binding proteins of H. somni. One of the surface proteins, IbpADR2 was shown to be protective for calves challenged after vaccination. The protective effect consisted of lowered clinical scores and lung lesion scores at necropsy. Protection was associated with specific IgG1 and IgG2 with low IgE responses. Iowa developed plasmids carrying the oriC region of Mycoplasma bovis. The plasmids were shown to replicate independently when introduced into M. bovis under antibiotic pressure, and could be removed by releasing the mycoplasma from antibiotic pressure during growth. This genetic tool is critically needed for the development of a modified live M. bovis vaccine. A saturation library of transposon mutants was developed from a virulent M. bovis strain. The mutants are used on ongoing studies to develop a modified live vaccine. Kansas studied weekly incidence of BRD morbidity risk in pens of feedlot cattle. They confirmed that mixed gender groups, multiple source groups, cattle that were shipped for long distances, and light weight calves were at increased risk of morbidity. A study comparing day one methaphilactic use of tulathromycin versus tilmicosin showed that the calves given the longer acting tulathromycin treatment had better health and performance parameters. Michigan has started a regional (upper Michigan peninsula) voluntary eradication campaign. Initial response and participation in the study has been very positive. South Dakota studied the impact of selenium and vitamin E supplementation on antibody response to BHV-1 vaccination and ovalbumin immunization of day-old dairy calves. Organic selenium administration improved overall immune responses of the calves. Wisconsin continued studies on the relationship between air quality, other environmental risk factors and dairy calf respiratory health.<br /> <br /> <br />

Publications

JOURNAL PUBLICATIONS:- <br /> <br /> <br /> Gershwin LJ, Gunther RA, Hornof WJ, Larson RF. Effect of infection with bovine respiratory syncytial virus on pulmonary clearance of an inhaled antigen in calves. Am J Vet Res 3:416-422, 2008.<br /> <br /> <br /> Geertsema RS, Worby C, Kruger RP, Tagawa Y, Russo R, Herdman DS, Lo K, Kimball RA, Dixon J, Corbeil LB. Protection of mice against H. somni septicemia by vaccination with recombinant immunoglobulin binding protein subunits. Vaccine 35:4506-4512, 2008.<br /> <br /> <br /> Reber AJ, Donovan DC, Gabbard J, Galland K, Aceves-Avila M, Holbert KA, Marshall L, Hurley DJ. Transfer of maternal colostral leukocytes promotes development of the neonatal immune system. II. Effects on neonatal lymphocytes. Vet. Immunol Immunopath 123:305-313, 2008.<br /> <br /> <br /> Reber AJ, Donovan DC, Gabbard J, Galland K, Aceves-Avila M, Holbert KA, Marshall L, Hurley DJ. Transfer of maternal colostral leukocytes promotes development of the neonatal immune system. II. Effects on monocyte lineage cells. Vet Immunol Immunopath 123:186-196, 2008.<br /> <br /> <br /> Chase CCL, Hurley DJ, Reber AJ. Neonatal immune development in the calf and its impact on vaccine response. Vet Clin North Am Food Anim Pract 24:87-104, 2008.<br /> <br /> <br /> Funk RA, Thomson DU, Renter DG, White BJ, Guillosou S. Comparison of pooled testing to individual testing with an ear-notch antigen capture ELISA testing for bovine viral diarrhea virus. Bovine Practitioner 42:93-97, 2008.<br /> <br /> <br /> Funk RA, Thomson DU, White BJ, Renter DG, Rust RR. Effects of ear-notch sample handling on reliability of antigen-capture ELISA attesting for BVDV. Bovine Practitioner 42:45-49, 2008.<br /> <br /> <br /> Nickell JS, White BJ, Larson RL, Blasi DA, Renter DG. Comparison of short term health and performance effects related to prophylactic administration of either tulathromycin or tilmicosin to beef stocker calves. Vet Therapeut 9:147-156, 2008.<br /> <br /> <br /> Sanderson MW, Dargatz DA, Wagner BA. Risk factors for initial respiratory disease morbidity counts in US feedlots. Canad Vet J 49:373-378, 2008.<br /> <br /> <br /> Muhammad-Tahir Z, Alocilja EC, Grooms DL. Indium tin oxide polyaniline biosensor: fabrication and characterization. Sensors 7:1123-11490, 2007.<br /> <br /> <br /> Jin JH, Zhang D, Alocilja EC, Grooms DL. Label-0free DNA sensor on nano-porous silicon-polypyrrole chip for monitoring Salmonella species. IEEE Sensors J 8:891-8954, 2008.<br /> <br /> <br /> Nanduri B, Lawrence ML, Peddinti DS, Burgess SC. Effects of sub-minimum inhibitory concentrations of antibiotics on the Pasteurella multocida proteome: a systems approach. Comparative ands Functional Genomics e-pub 254836, 2008.<br /> <br /> <br /> Pinchuk GV Lee S-R, Nanduri B, Honsiger KL, Stokes JV, Pinchuk LM. BVDV differentially alter the expression of the protein kinases and related proteins affecting the development of infection and anti-viral mechanisms in bovine monocytes. Biochim Biophys Acta 1784: 1234-1247, 2008.<br /> <br /> <br /> Lee S-R, Pharr GT, Boyd BL, Pinchuk LM. BVDV modulate toll-like receptors, cytokines and co-stimulatory molecule gene expression in bovine PBMC. Comp Immun Microbiol Infect Dis 31:403-418, 2008.<br /> <br /> <br /> Perez S, Meyer F, Saira K, Doster A, Jones C. Premature expression of the latency related (LR) RNA correlates with higher levels of beta-interferon RNA expression in productively infected cells. J Gen Virol 89:1338-1345, 2008.<br /> <br /> <br /> Geiser V, Rose S, Jones C. BHV-1 induces cell death by a cell type dependent fashion. Microb Pathogen 44:459-466, 2008.<br /> <br /> <br /> Liu ZF, Brum MCS, Doster A, Jones C, Chowdhury SI. A BHV-1 mutant virus specifying a carboxyl terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves. J Virol 82:7432-7442, 2008.<br /> <br /> <br /> Brum MCS, Coats C, Sangena BR, Doster A, Jones C, Chowdhury SI. BHV-1 anterograde neuronal transport from trigeminal ganglia to nose and eye requires glycoprotein E. In Press, J Neurovirol.<br /> <br /> <br /> Zimmerman AD, Butterbaugh RE, Herbert JM, Haas JM, Frank NE, Luempert LG, Chase CCL. Efficacy of BHV-1 inactivated vaccine against abortion and stillbirth in pregnant heifers. JAVMA 231:1386-1389, 2007.<br /> <br /> <br /> Ridpath JF, Mark CS, Chase CCL, Ridpath AC, Neill JD. Febrile response and decrease in circulating lymphocytes following acute infection in white tail deer fawns with either BVDV1 or a BVDV2 strain. J Wildlife Dis 43:653-659, 2007.<br /> <br /> <br /> Atapattu DN, Czuprynski CJ. Mannheimina haemolytica leukotocxin binds to lipid rafts of bovine lymphoblastoid cells (BL-3) and is internalized in a dynamin-2 and clathrin-dependent manner. Infect Immun 75:4719-4725.<br /> <br /> <br /> Behling-Kelly E, McClenahan D, Kim KS, Czuprynski. Viable Haemophilus somnus induce myosin light chain kinase-dependent decrease in brain endothelial cell monolayer resistance. Infect Immun 75:4572-4581.<br /> <br /> <br /> Behling-Kelly E, Kim KS, Czuprynski CJ. Haemophilus somnus activation of brain endothelial cells: potential role for local cytokine production and thrombosis in CNS infection. Thromb Haemostas 98:823-830, 2007.<br /> <br /> <br /> Kuckleburg CJ, McClenahan DJ, Czuprynski CJ. Platelet activation by Histophilus somni and its LOS induces endothelial cell pro-inflammatory cytoikine responses and platelet internalization. Shock 29:189-196, 2008.<br /> <br /> <br /> Kuckleburg CJ, Tiwari R, Czuprynski CJ. Endothelial cell apoptosis induced by bacteria-activated platelets requires caspase-8 and -9 and generation of reactive oxygen species. Thromb Haemost 99:363-372, 2008.<br /> <br /> <br /> McClenahan D, Hellebrand K, Atapattu D, Aulik N, Carlton S, Kapuir A, Czuprynski CJ. Effects of LPS and Mannheimina haemolytica leukotoxin on bovine lung microvascular endothelial cells and alveolar epithelial cells. Clin Vaccine Immunol 15:338-347, 2008.<br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> REVIEWS AND BOOK CHAPTERS:-<br /> <br /> <br /> Jones C, Chowdhury SI. A review of the biology of BHV-1. Its role as a cofactor in the BRD complex, and development of improved vaccines. Adv An Health 8:187-205, 2008.<br /> <br /> <br /> Behling-Kelly E, Czuprynski CJ. Endothelial cells as active participants in veterinary infections and inflammatory disorders. Anim. Health Res Rev 8:47-58, 2007.<br /> <br /> <br /> Rosenbusch, R. F. Chapter 43: Mycoplasmas in bovine respiratory disease. In Current Veterinary Therapy Food Animal Practice Vol. 5, D. E. Anderson and D. M. Rings, eds., Saunders, St. Louis, 2009.<br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> ABSTRACTS AND CONFERENCE PROCEEDINGS:-<br /> <br /> <br /> Haase C, Quesada M, Mukhtar E, Sharp P, Larson LJ, Thomas C, Schultz RD. Comparative study of three major modified live bovine vaccines for BVDV, BHV-1, PI-3, BRSV, and 5 Leptospira serovars. 88th CRWAD meeting, Dec 2-4, 2007, Chicago. IL.<br /> <br /> <br /> Reber AJ, Donovan DC, Gabbard J, Gallanfd K, Aceves-Avila M, Holbert KA, Marshall L, Hurley DJ. Transfer of maternal colostral leukocytres promotes development of neonatal lymphocytes. 88th CRWAD meeting, Dec 2-4, 2007, Chicago. IL.<br /> <br /> <br /> Woolums AR. Vaccination of calves  New information on the effects of maternal antibody. 40th AABP convention, Sep 20, 2007, Vancouver, BC.<br /> <br /> <br /> Rosenbusch RF, Lee N, Christensen L. Identification of Mycoplasma bovis genes involved in uptake of unopsonized mycoplasmas by activated macrophages. 17th International Organization for Microbiology Congress, Tianjin, China, 2008.<br /> <br /> <br /> Rosenbusch RF, Lee N, Zamzow DR, Riedell CR, Christensen L. Use of a respiratory tract infection model in cattle to screen transposon 4001 mutants of Mycoplasma bovis. 17th International Organization for Microbiology Congress, Tianjin, China, 2008.<br /> <br /> <br /> Lee N, Zamzow DR, Riedell, CR, Rosenbusch RF. Development of a transposon 4001 mutant library of a pathogenic Mycoplasma bovis strain. 17th International Organization for Microbiology Congress, Tianjin, China, 2008.<br /> <br /> <br /> Rosenbusch RF, Zamzow DR, Riedell CD, Lee. N Construction of a random mutant <br /> library of Mycoplasma bovis. 108th General Meeting of the American Society for Microbiology, Boston, 2008.<br /> <br /> <br /> Lee N, Zamzow DR, Rosenbusch RF. Identification of the origin of replication in the chromosome of Mycoplasma bovis and its use to construct a replicative plasmid. 108th General Meeting of the American Society for Microbiology, Boston, 2008. <br /> <br /> <br /> Berghaus LJ, Vandenplas ML, Galland KL, Davis CE, Hurley KAE, Blas-Machado U, Woolums AR. Comparison of 3 methods of identification of nasa; BRSV shedding by calves. 88th CRWAD meeting, Dec 2-4, 2007, Chicago. IL.<br /> <br /> <br /> Woolums AR. The problem of insufficient funding for cattle health research. Presented to the AABP Board of Governors Annual Meeting, March 2008.<br /> <br /> <br /> Woolums AR. Immune development in the neonatal calf. Minnesota Nutrition satellite conference, Sep 16, 2008, Owatonna, MN.<br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> OTHER (M.S. Thesis):-<br /> <br /> <br /> Rivera, Jose. Effects of BHV-1 infection on bovine neutrophil adhesion to bronchial epithelial cells in vitro. MS thesis, Madison, WI.<br /> <br /> <br />

Impact Statements

1. OBJECTIVE 1: To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents. Evaluation of BVDV ear-notch testing methodologies by KS and OK will lead to improved diagnostic techniques that will be useful for diagnostic laboratories, veterinarians and producers. Further development of rapid chute-side tests for BRD and BVDV persistence (KS, MI) will provide new tools for practitioners. Continued surveillance of BRD cases by diagnostic laboratories provides updated information on prevalence rates and shifts in BRD ecology, and this is important information for researchers, diagnostic laboratories, the veterinary industry and veterinarians.
2. OBJECTIVE 2: To investigate the basic biology, molecular pathogenesis, and immunopathogenesis of polymicrobial infections including important viral and bacterial agents. In vitro studies of BRD pathogens interacting with host cells (GA, KS, NE, WI) will lead to the formulation of new strategies for diagnosis, prevention, and therapy of BRD. Efforts to understand how pathogens modulate their virulence attributes (CA, MS, NE) are important in developing knowledge necessary for the development of control strategies. Finally, understanding the interaction between BRD pathogens (CA) can lead to better prevention and therapy practices. While the advances for this objective are more basic in nature, they have the potential to impact a much wider constituency of stakeholders, including the consumer public.
3. OBJECIVE 3:To develop management and prevention strategies that incorporate new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss. Several approaches to improved vaccines are being investigated. These include improved H. somni (CA), improved M. bovis (IA) and improved BHV-1 (NE) vaccines. Environmental and management factors and their impact on BRD continue to be studied (KS, WI) with a focus on economical alternatives to reduce negative effects and potentiate beneficial effects. This information is important to veterinarians and producers. A start on eradication of BVDV from the USA has been made with a regional project (MI).

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Report Information

Participants

Georgia (Woolums, Amelia, awoolums@uga.edu); Iowa (Rosenbusch, Ricardo, rfrosenb@iastate.edu); Michigan (Grooms, Dan, groomsd@cvm.msu.edu); Nebraska (McVey, DS, dmcvey2@unl.edu); Oklahoma (Fulton, RW, rfulton@okstate.edu); South Dakota (Chase, Christopher, Christopher.Chase@sdstate.edu); Wisconsin (Czuprynski, Charles, czuprync@svm.vetmed.wisc.edu);
Guests: Peter Johnson (USDA CSREES), Eileen Thacker (USDA ARS), Bob Briggs (USDA NADC), Maria Prado (UTenn)

Brief Summary of Minutes

NC-1027 An Integrated Approach to the Control of Bovine Respiratory Disease
Annual Report, 2009

Annual Meeting: In conjunction with 2009 BRD Symposium, Colorado Springs, CO; August 6, 2009.

Stations reporting: Georgia, Iowa, Kansas, Michigan, Nebraska, Oklahoma, South Dakota, Wisconsin
Stations attending: Georgia (Woolums), Iowa (Rosenbusch), Michigan (Grooms), Nebraska (McVey), Oklahoma (Fulton), South Dakota (Chase), Wisconsin (Czuprynski)
Guests: Dr. Peter Johnson (USDA CSREES), Dr. Eileen Thacker (USDA ARS), Dr. Bob Briggs (USDA NADC), Dr. Maria Prado

Agenda:

7:30 PM Welcome
7:35 PM Discussion on BRDC meeting and outcomes from the meeting and Open Forum at AABP
8:35 PM Future of NC1027-
9:30 PM Adjourn

Discussion of the BRD Symposium which ended this evening. Consensus is that meeting was a great success. Many positive comments from attendees to membership. Dr. Woolums will talk to Dr. Gyles of Animal Health re publication of proceedings. Options for profit: will try to put over in rolling account at University of Georgia. If this is a problem, may keep money with Dr. Jim Roths organization; outside of Iowa State. BVD group did this with their profit.

What to do with profit: Dr. Fulton suggests organizing committee should get what speakers got (for reimbursement and honorarium). Once bills are paid will discuss options further, possibilities include student research awards at national meetings, or save money for next Symposium.

Breakout session summaries: Dr. Rosenbusch will summarize. Where to put it? Would be ideal to keep with proceedings in AHRR if Dr. Gyles will accept it. That will be our first plan. Were planning a bulleted list; AHRR may want different format.

Proceedings: some still available. Will make available on website for $15. Will charge the same for pdf on CD.

Open Forum on BRD at AABP: was a big success last year; about 60-70 AABP attendees came to have informal discussion. Drs. Chase, Grooms, Woolums are on new Bovine Respiratory Disease Committee, which is scheduled to meet Thur AM of AABP meeting. Dan points out that much competition for that time. Dan Grooms will talk to Gatz Riddell re possibility of meeting at another time. Will ask Gatz if we can put flyer in registration packets as in past.


Discussion of future of NC-1027: consensus was that poor attendance in recent years has been disappointing. Discussion of possibilities of terminating group and founding a Ruminant Respiratory Disease Society. Consensus that no great loss if group ended since very few get any money for involvement any more, and those that do will be able to get money by other routes through their AES. Downside to ending project: we may have no more impetus to meet, which is good opportunity for sharing research info and discussing research collaboration possibilities.

Dr. Peter Johnson offered to require AFRI awardees funded for BRD research to present annually in conjunction with our annual meeting as a Society or other format. This would help maintain drive for scientific presentations. Some discussion of annually meeting in conjunction with CRWAD, but majority felt that good to meet with AABP because this gives researchers opportunity to interact with private practitioners, to possible benefit of both parties.

Dr. Johnson reminded us that if we maintain multistate project we can invite with reason anyone to join committeedo not have to be only people at AESs. Even those from outside of U.S. OK; Dr. Hahn would have to approve but Dr. J thinks probably not a problem.

Dr. Chase will talk to his AES director about pros/cons and logistics of terminating project early and will report back.

If we do terminate, plan to pursue Ruminant Respiratory Disease Society. Plan for meeting: meet next year at AABP. Come up with new name. Send out invitation to all interested parties. Tell them to meet at AABP. AFRI will let BRD awardees know they need to present in conjunction with our meeting. Remember if we are a multistate next year we can still invite anyone we want. Some discussion: what to have AFRI awardees do, poster or presentation? Many felt presenting at AABP poster session would be best. Consensus is that we should have 1) meeting of society at AABP; 2) ask AABP for a recurring block for oral presentations in Research Session, and have a selected number of AFRI awardees present as well as some Society members; 3) rest of AFRI awardees present poster

Peter Johnson will tell AFRI awardees to block off Aug 18-22 and plan to present there; more details to follow

Update re CAP from Dr. Johnson: there will be new CAP project funded, RFP will be in 2010, will be openmight identify certain diseases or issues, but could be broad, too. Could request individuals to put in applications, then some internal committee would decide which to put forth. Will only have one at a time, may have every other year. How much/durationmaybe 1.5 million for 4 years. This would fit well with budget.
Remember re CAP: project director has to dedicate 35-40% of their time to managing CAP, especially in first year. Need administrative assistant with a good salarynot a $15k/y person! Project director needs to have even keeled personality, someone who community trusts. Ideally they would not receive any money except for salary support so those who apply for funding in CAP dont think they have conflict of interest. Remember project has to be integratedfrom d. 1, extension has to be an objective; need education component, too (sounds like this is part of outreach). Have an extension specialist who is co-PD and who manages that objective. So have research team and outreach team and both have to be strong. No more than 2/3 of budget can go to any one of these elements. Have an external scientific review board WHO WILL NOT BE FUNDED BY CAP review the submitted proposals. Then no hard feelings between members and project director.
Q: can education be at any level? Yes, even 4-H involvement can work!
Challenging thing is that community is large, but not everyone can get money. They do have developmental grants every year so new people can join. Can talk to PRRSV people for info. Remember each year there is a reviewscientific board reviews progress periodically. PRRSV and Johnes: stakeholder board has final say. So far they havent vetoed anything.
Q: could former CAP members apply for something new (new disease)? Yes.

Is one of the current CAPs ending? No.

Definitely look at how PRRSV and Johnes have worked. PRRSV CAP 2 is different than PRRSV CAP 1. Johnes is very different.

Middle of Nov is target AFRI RFI, letters of intent will probably be due in Jan 10; deadlines for upcoming year will probably be similar to last year.

Accomplishments

Accomplishments:<br /> <br /> Objective 1: To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents.<br /> <br /> Identification of various BRSV strains by commercial human RSV diagnostic kits: Georgia has provided BRSV isolates for testing in this project being carried out by Michigan. <br /> The utility of physiologic, behavioral and pathological changes as objective indicators of early respiratory disease in calves with experimental M. haemolytica pneumonia were examined. Temperature, pulse, respiration, clinical illness score, activity level (pedometers and accelerometers) respiratory character, arterial blood gas, blood chemistry and complete blood counts were assessed. Routine laboratory parameters and subjective measures were not reliable indicators of progression of pneumonia. However activity, objectively measured with pedometers and accelerometers, appears to be a promising tool for early recognition of bovine respiratory disease. <br /> BVDV PI Cattle Monitoring in the Feedyard; Surveillance for the PI cattle entering a southwest Kansas feedlot has continued since July 1, 2004 through November 2008. The PI cattle based on the positive tests of ACE and IHC from ear notches had serums collected, virus propagated in cell culture and sequenced with BVDV subtypes determined. Since 2004 through 2008, 1280 PI cattle have had the viruses subtyped with the distribution of subtypes: BVDV1b, 1003 (78.4%); BVDV1a, 154 (12.0%); and BVDV2a, 123 (9.6%). The USDA, ARS, NADC provided extensive collaboration by sequencing the viruses. OSU also provided an OSU monoclonal antibody to BVDV to the USDA laboratory of Dr. Julia Ridpath.<br /> Multiple diagnostic tests to identify cattle with bovine viral diarrhea virus and duration of positive test results in persistently infected cattle: Several tests for bovine viral diarrhea virus (BVDV) were applied to samples collected from twelve persistently infected (PI) cattle with BVDV subtypes of U.S. cattle: BVDV1a, BVDV1b, and BVDV2a. These collections were made monthly from December 20, 2005 through November 27, 2006 (day 0 to day 342). The samples collected included clotted blood for serums, nasal swabs, and fresh and formalin-fixed ear notches. The tests utilized included viral titration of nasal swabs and serums for infectious virus; antigen-capture ELISA (ACE) on fresh ear notches, serums, and nasal swabs; gel-based polymerase chain reaction (PCR) on serums, nasal swabs, and fresh ear notches; immunohistochemistry (IHC) on formalin-fixed notches; and serology for BVDV antibodies in serums. Of the twelve animals starting the study, three died with Mucosal disease. The IHC and ACE tests on ear notches were positive throughout the study as were the serums for ACE and PCR. There was detectable virus in nasal swabs of all cattle throughout the study with the exception of a few that were toxic to cell cultures. The serums had viral titers e log10 1.6 in all samples of all PI cattle, except for three collections of one animal. PCR testing of individual samples were most often positive, although there were several with equivocal results. The BVDV antibodies were due to vaccination or exposure to heterologous PI strains, and did not appear to interfere with any BVDV test. These studies illustrate that PI cattle may be identified by several tests, yet differentiation of PI cattle from acute BVDV infections of cattle will require additional testing, especially for blood samples and nasal swabs positive on initial test. Also, calves PI with BVDV are continual shedders of infectious virus as shown by infectivity of nasal swab samples over the 11-month study.<br /> Evaluation of economic effects and the health and performance of the general cattle population after exposure to cattle persistently infected with bovine viral diarrhea virus in a starter feedlot: The objectives of the study were to investigate the economic effects and health and performance of the general cattle population after exposure to cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV) in a feedlot. There were 21,743 high-risk calves from the southeastern United States in a commercial feedlot. The PI status was determined by use of an antigen-capture ELISA (ACE) and confirmed<br /> by use of a second ACE, reverse transcriptasePCR assay of sera, immunohistochemistry of skin samples for BVDV antigen, and virus isolation from sera. Groups with various amounts of exposure to BVDV PI cattle were used. After being placed in the feedlot, identified PI cattle were removed from 1 section, but PI cattle remained in another section of the feedlot. Exposure<br /> groups for cattle lots arriving without PI animals were determined by spatial association to cattle lots, with PI animals remaining or removed from the lot.<br /> There were 15,348 cattle that maintained their exposure group. Performance outcomes improved slightly among the 5 exposure groups as the risk for exposure to BVDV PI cattle decreased. Health outcomes had an association with exposure risk that depended on the exposure group. Comparing cattle lots with direct exposure with those without direct exposure revealed significant improvements in all performance outcomes and in first relapse percentage and mortality percentage in the health outcomes. Economic analysis revealed that fatalities<br /> accounted for losses of $5.26/animal and performance losses were $88.26/animal.<br /> This study provided evidence that exposure of the general population of feedlot cattle to BVDV PI animals resulted in substantial costs attributable to negative effects on performance and increased fatalities.<br /> In South Dakota and Wisconsin, the infectious agents associated with bovine respiratory disease complex were monitored by bacterial culture, virus isolation, and fluorescent antibody techniques.<br /> <br /> Objective 2: To investigate the basic biology, molecular pathogenesis, and immunopathogenesis of polymicrobial infections including important viral and bacterial agents. <br /> <br /> Development of primary bovine bronchial epithelial cell (BBEC) cultures from cells taken from living cattle by bronchial brushing: This project is slowly ongoing. <br /> The in vitro effects of substance P alone or in combination with TNF, LPS, and/or histamine on bovine endothelial cell cultures were investigated. Permeability of endothelial cultures was determined by diffusion of albumin bound to Evans Blue dye through and endothelial monolayer cultured on a permeable membrane. The permeability of microvascular endothelium increased the greatest after stimulation with a combination of substance P and histamine. Responses to substance P and histamine alone were less than the combination, but equal to each other. Permeability of macrovascular endothelium increased the greatest after stimulation with a combination of substance P and LPS. The permeability induces by a substance P/histamine combination was slightly less than that produced by substance P/LPS, and the least stimulation occurred with substance P, TNF and LPS alone.<br /> Functional analysis of bICPO, a bovine herpesvirus 1 gene that is a promiscuous transactivator: bICP0 can activate expression of all three classes of viral genes, is expressed throughout productive infection, and is thus considered to be the most important viral regulatory gene. We have demonstrated that a C3HC4 zinc ring finger near the amino terminus of bICP0 plays an important role in activating transcription and productive <br /> infection. Furthermore, bICP0 interacts with chromatin remodeling enzymes {histone deacytlase 1 (HDAC1) and a histone acetylase (p300)}. Recent studies have demonstrated that bICP0 also inhibits interferon dependent transcription, suggesting that bICP0 regulates innate immune responses. <br /> Regulation of the latency-reactivation cycle by the bovine herpesvirus 1 latency related (LR) gene: Like other members of this subfamily, a latent infection is established in sensory neurons following acute infection. However, the virus can reactivate and spread to other cattle. Reactivation from latency is the mechanism by which the virus survives in nature and is <br /> thus an important property of pathogenesis. During a latent infection, one abundant viral transcript can be detected; the latency related RNA (LR-RNA). Expression of LR proteins is necessary for reactivation from latency and protecting infected neurons from apoptosis. We have demonstrated that a LR encoded protein (ORF-2) or ORF-2 fusion proteins encoded by alternatively spliced LR transcripts inhibit cold shock or Fas ligand induced apoptosis in mouse neuroblastoma cells (neuro-2A). Frame shift mutants of ORF-2 do not inhibit apoptosis suggesting that protein expression, not LR-RNA expression, mediates the anti-apoptosis activity of the LR gene in transfected neuro-2A cells. Yeast two-hybrid analysis has revealed that ORF-2 sequences specifically interact with two cellular proteins that regulate cell survival and differentiation (Notch 1 and Notch 3). We have also identified sequences in the LR gene that inhibit bICP0 expression. It appears that a viral encoded micro-RNA may inhibit bICP0 expression. <br /> Mannheimia haemolytica immunity of cattle following vaccination with chimeric protein: This study was done to determine if intranasal vaccination of young Holstein calves with a chimeric protein containing the immunodominant surface epitope of Mannheimia haemolytica PlpE (R2) and the neutralizing epitope of leukotoxin (NLKT) in the presence cholera toxin (CT) could stimulate secretory and systemic antibodies against M. haemolytica antigens. Dairy calves ranging from 3 to 18 days old were intranasally vaccinated with PBS, 10 or 100 µg of M. haemolytica chimeric protein SAC89, which contains two copies each of R2 and of NLKT, with 20 µg of CT. Vaccination with either concentration of SAC89 stimulated significant (p < 0.05) increases in intranasal IgA antibodies to PlpE and to LKT, whereas significant increases (p < 0.05) in serum antibodies were only demonstrated against M. haemolytica whole bacterial cells in the 10 µg SAC89 concentration group. Therefore, intranasal vaccination with R2-NLKT chimeric protein plus CT enhanced mucosal antibody responses to M. haemolytica PlpE and LKT. Further studies are needed to determine if this intranasal vaccination is a viable method for stimulating protection of young dairy calves against M. haemolytica.<br /> The effect of BVDV on innate immunity in the developing fetus: This project is part of a collaborative project between South Dakota and Dr. Thomas Hanson of Colorado State University. We have developed a method to access macrophage activity in the fetal calf using Kueffer Cells from the liver. These studies are looking at fetuses at different gestarion lengths and the effect on innate immunity.<br /> The effect of BVDV on macrophage cytoskeleton. This project has just begun but is using 5 different NCP BVDV strains and determining their effect on actin and microtubule architecture.<br /> During the past year Wisconsin ontinued to investigate intracellular pathways that result in bovine lymphoblastoid (BL3) cell death following exposure to M. haemolytica LKT. Experiments confirm that LKT is also transported to mitochondria in bovine peripheral blood neutrophils and monocytes. However we find some differences in what mitochondrial proteins bind LKT, and in the ability of cyclosporine to prevent LKT binding to mitochondria. A student in the lab designed an LKT construct to track internalization and transport of LKT in leukocytes. This effort was aided by Dr. Maheswaran at Minnesota. We also reported that brief heating of LKT increases its cytotoxic potency for bovine leukocytes and eliminates the strict species-specific toxicity of the native toxin. Heating of the related RTX toxin, the E. coli hemolysin, similarly increases its cytotoxicity for target cells. <br /> It is clear from both field observations and experimental studies that active infection with BHV-1 or other bovine respiratory viruses predisposes cattle to more severe pneumonia with M. haemolytica. We have evidence that infection of primary bovine bronchiolar epithelial cell lines with BHV-1 increases the adhesion of M. haemolytica to these cells. Likewise it also increases the adherence of bovine neutrophils to the viral-infected epithelial cells. Conditioned medium from BHV-1 infected epithelial cells increases neutrophil adhesion to uninfected epithelial cells. We also find that BHV-1 infection increases epithelial cell expression of IL-8, and several inflammatory cytokines (IL-1a, IL-1b, TNF-a). We showed that conditioned medium from BHV-1 infected epithelial cells is chemotactic for bovine PMNs, and activates PMN oxidative burst and expression of CD11a/CD18. The latter is associated with increased killing by M. haemolytica LKT. These studies have been assisted by scientists at the Nebraska Medical Center. <br /> Wisconsin compared the responses of bovine pulmonary epithelial and endothelial cells to LPS and LKT. Endothelial cells are more responsive to LPS, exhibiting changes in transepithelial electrical resistance (TEER), expression of inflammatory cytokines, and morphologic and biochemical changes consistent with apoptosis. Epithelial cells also produced inflammatory cytokines in response to LPS, but did not demonstrate decreased TEER or cytotoxic changes, unless activated neutrophils were included in the incubation mixture. Both epithelial and endothelial cells had similar TLR-4 mRNA levels, suggesting that other differences in signaling are responsible for the dichotomy in response to LPS between the two cell types. ATP has also been examined as a potential mediator of changes to respiratory epithelial and endothelial cells. Our results show that ATP causes rapid but reversible morphologic changes in epithelial but not endothelial cells. In contrast, both epithelial and endothelial cell monolayers exhibit increased permeability as measure by electrical conductance TEER) following incubation with micromolar concentrations of ATP in vitro. <br /> Wisconsin also continued their studies of mechanisms by which Haemophilus somnus causes vasculitis. The most severe manifestation of H. somnus infection is thrombomeningoencephalitis (TME). Cerebral microvascular endothelial cells are the primary cellular constituent of the blood brain barrier, which protects the central nervous system from inflammation and infectious agents. Recent results indicated that incubation of brain microvascular endothelial cells with H. somnus increases their surface expression of PECAM-1 and eNOS. We hypothesize that these events might contribute to platelet and neutrophil adhesion to endothelial cellsPECAM-1) and increased vascular permeability (eNOS). Consistentwith this inferecen we show that PMN mibration across endothelial monolayers is inhibited by anti-PECAM antibodies.The observed alterations in the BBECs could play a role in the development of vasculitis, thrombosis and inflammation that result in blood brain barrier dysfunction and bacterial invasion of the brain parenchyma. These studies involved collaboration with Dr. Corbeil (California) and Dr. K.S. Kim (Johns Hopkins).<br /> <br /> Objective 3: To develop management and prevention strategies that incorporate new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss.<br /> <br /> Evaluating the effect of vaccinating calves in the face of maternal antibody (IFOMA) on subsequent response to booster vaccination at weaning. The trial to determine whether a multivalent intranasal vaccine (Onset IN) can prime calves vaccinated at 3 days of life or at 60 days of life for an improved response to booster vaccination at weaning is underway. Development of small inhibiting RNA (siRNA) to minimize disease due to BRSV infection. This project is progressing. We recently showed that siRNA directed against the mRNA of the BRSV P gene decreased expression of P mRNA as measured by real time RT-PCR in BRSV infected cells; this confirmed that the siRNA acted as predicted. Initial results in cell culture indicate that siRNA treatment decreased viral replication, as predicted. Assessment of the results and repetition of the studies is underway to confirm that siRNA can suppress BRSV replication. <br /> IA had proposed to provide leadership in the development of a modified-live vaccine against Mycoplasma bovis during the project period. Extending its efforts to develop tools to genetically engineer this mycoplasma, mutants with single gene disruptions were studied for resistance to macrophage attack. Wild-type M. bovis strains typically resist macrophages, and this resistance plays a prominent role in the pathogenesis of pneumonia, polyarthritis, and tenosynovitis as caused by the pathogen. Two genes of M. bovis were identified as required to maintain this resistance to macrophage attack. <br /> The Genetics of Feedlot Health: South Dakota along with Texas, Illinois, Missouri, New York and Colorado are involved in the Genetics of Feedlot Health Project. This study looks at behavior, genetics, nutrition along with microbiology and immunology on respiratory disease and carcass quality. This 2-year project is in its final year and all the data is still being analyzed.

Publications

B.J. White, D.G. Renter. Bayesian estimation of the performance of clinical<br /> observations and harvest lung scores for diagnosing bovine respiratory<br /> disease in post-weaned beef calves. J Vet Diagn Invest July 2009 Vol. 21,<br /> Number 4: 446-453. <br /> <br /> A. Babcock, B.J. White, D.G. Renter, S.S. Dritz, D.U. Thomson. Feedlot health and performance effects associated with the timing of respiratory disease treatment. J Anim Sci 2009 87:314-227.<br /> <br /> G.A. Hanzlicek, B.J. White, D. Anderson, D. Mosier, D.G. Renter. Pathological and physiological changes following induced Mannheimia pneumonia in beef feeder calves. Amer J Vet Res. Accepted. <br /> <br /> B.J. White, G. Hanzlicek, M.W. Sanderson, D.E. Sanderson, R.L. Larson. Prevalence of Mollicutes species in beef feeder calves at arrival and first treatment for bovine respiratory disease. Can Vet J. Accepted.<br /> <br /> Shen, W. and C. Jones. 2008. Open reading frame 2 encoded by the latency related gene of bovine herpesvirus 1 has anti-apoptosis activity in transiently transfected neuroblastoma cells. J. Virology, 82:10940-10945. <br /> <br /> Saira, K., S. Chowdhury, N. Gaudreault, L. da Silva, G. Henderson, A. Doster, & C. Jones. 2008. The zinc RING finger of the bovine herpesvirus 1 encoded bICP0 protein is crucial for viral replication and virulence. J. Virology, 82:12060-12068. <br /> <br /> Brum. M.C.S., C. Coats, B.R. Sangena, A. Doster, C. Jones, and S.I. Chowdhury. <br /> 2009. Bovine herpesvirus type 1 (BoHV-1) anterograde neuronal transport from trigeminal ganglia to nose and eye requires glycoprotein E. J. Neurovirology, 15:1-6. <br /> <br /> Saira, K., Y. Zhou, and C. Jones. 2009. The infected cell protein 0 encoded by bovine herpesvirus 1 (bICP0) associates with interferon regulatory factor 7 (IRF7), and consequently inhibits beta interferon promoter activity. J. Virology. 83:3977-3981. <br /> <br /> Meyer, F. and C. Jones. 2009. C/EBP-alpha cooperates with bTIF to activate the bovine herpesvirus 1 immediate early transcription unit 1 promoter. J. Neurovirology 15:123-130. <br /> <br /> Ellis, J., S. Gow, N. Goji, C. Jones, A. Workman, G. Henderson, G. Alaniz, and T. Meinert. Efficacy of a combination viral vaccine in protecting cattle from experimental infection with bovine herpesviruses-1 isolated from recent vaccine breaks. J. of American Veterinary Medical Association, IN PRESS. <br /> <br /> Workman, A., S. Perez, A. Doster, and C. Jones. 2009. Dexamethasone treatment of calves latently infected with bovine herpesvirus 1 (BHV-1) leads to activation of the bICP0 early promoter, in part by the cellular transcription factor C/EBP-alpha. J. Virology, IN PRESS. <br /> <br /> Taylor JD, Fulton RW, Dabo SM, Lehenbauer TW, Step DL, Confer AW. Molecular characterization of Pasteurella multocida isolates from fatal cases of bovine respiratory disease. Can Vet J, 2009, in press<br /> Taylor JD, Fulton RW, Lehenbauer TW, Step DL, Confer AW. The epidemiology of bovine respiratory disease: What is the evidence for preventive measures? Can Vet J, 2009, in press<br /> Confer AW, Ayalew S, Step D L, Trojan B, Montelongo M. Intranasal vaccination of young Holstein calves with Mannheimia haemolytica chimeric protein PlpE-LKT (SAC89) and cholera toxin Vet Immun & Immunopathol, 2009, available online as of May 23, 2009<br /> Singh K, Confer AW, Hope JC, Rizzi T, Wyckoff JW, Weng HY, Ritchey JW. Cytokine mRNA modulation in bovine alveolar macrophages challenged with the wild type and leukotoxin-deficient mutant of Mannheimia haemolytica. Mol Pathog, accepted with revisions, 2009 <br /> Ayalew S, Step DL, Montelongo M, Confer AW. Intranasal vaccination of calves with Mannheimia haemolytica chimeric protein containing the major surface epitope of outer membrane lipoprotein PlpE, the neutralizing epitope of leukotoxin, and cholera toxin subunit B. Vet Immun & Immunopathol, In press, 2009.<br /> Step, D.L., Krehbiel, C.R., DePra, H.A., Cranston, J.J., Fulton,R.W.,Kirkpatrick,J.A., Gill, D.R., Payton, M.E., Montelongo, M.A., Confer, A.W.: Effects of Commingling Beef Calves from Different Sources and Weaning Protocols During a Forty-Two-Day Receiving Period on Performance and Bovine Respiratory Disease. Journal of Animal Science. 86:1-13, 2008.<br /> Step, D.L., Litherland, N.B., Burciaga-Robles, L.O., Breshears, M.A., Krehbiel, C.R., Confer, A.W., Fulton, R.W., Morgan, G.L., Thornsberry, R.M., Fassig, S.M.: Clinical Observations, Biochemical Data, and Postmortem and Histopathologic Findings in Young Dairy Calves Fed Zeolite Clinoptilolite Binder Combined with Milk Replacer. American Journal for Veterinary Research 69: 1587-1594, 2008.<br /> <br /> Hessman, B.E., Fulton, R.W., Sjeklocha, D.B., Murphy, T.A., Ridpath, J.F., Payton, M.E.: Evaluation of the Economic Effects and the Health and Performance of the General Cattle Population After Exposure to Cattle Persistently Infected with Bovine Viral Diarrhea Virus in a Starter Feedlot. American Journal for Veterinary Research. 70:73-85, 2009.<br /> <br /> Fulton, R.W., Hessman, B.E., Ridpath, J.F., Johnson, B.J., Burge, L.J., Kapil.S. Braziel, B., Kautz, K., Reck, A.: Multiple Diagnostic Tests to Identify Cattle with Bovine Viral Diarrhea Virus and Duration of Positive Tests in Persistently Infected Cattle. In Press. Canadian Journal for Veterinary Research. 73: 117-124, 2009.<br /> <br /> Fulton, R.W., Whitley, E.M., Johnson, B.J., Ridpath, J.F., Kapil, S., Burge, L.J., Cook, B.J., Confer, A.W.: Beef Herds in a South Central United States Study: Prevalence of Bovine Viral Diarrhea Virus (BVDV) Persistently Infected Cattle and BVDV Subtypes in Affected Cattle. In press. October 2009. Canadian Journal for Veterinary Research<br /> <br /> Step, D.L., Krehbiel, C.R.., Burciaga-Robles, LO., Holland, B.P., Fulton,R.W., Confer,A.W., Hutcheson, J., Brister, D., Newcomb,H., Bechtol, D.: A Comparison of Single Vaccination and Revaccination with a Modified Live BHV1-BVDV( type 1 and 2-PI3V-BRSV Vaccine in the Prevention of Bovine Respiratory Disease. Accepted for publication .2009. Journal of American Veterinary Medical Association<br /> <br /> Ridpath, J.F., Fulton, R.W.: Knowledge Gaps Impacting the Development of BVDV Control Programs in the United States. 2009. Accepted for publication. 2009. Journal of American Veterinary Medical Association.<br /> <br /> Fulton, R.W., Blood, K.S., Panciera, R.J., Payton, M.E., Ridpath, J.F., Confer, A.W., Saliki, J.T., Burge, L.J., Welsh, R.D., Johnson, B.J., Reck. A.: Lung Pathology and Infectious Agents in Fatal Feedlot Pneumonias and Relationship with Mortality, Disease Onset, and Treatments. Journal of Veterinary Diagnostic Investigation 21: July 2009, in press.<br /> <br /> Chase CCL, LJ. Braun, P Leslie-Steen, T Graham, D Miskimins JF Ridpath. 2008. Bovine Viral Diarrhea Virus Multi-Organ Infection in Two White-tailed Deer in Southeastern South Dakota. J Wildlife Dis 44:753-759<br /> <br /> Ridpath JR, EA Driskell, CCL Chase, JD Neill, MV Palmer, BW Broderson. Reproductive tract disease associated with inoculation of pregnant white-tailed deer with bovine viral diarrhea virus. AJVR 69:1630-1636.<br /> <br /> Zimmerman AD, RE Buterbaugh, JA Schnackel, CCL Chase. Efficacy of a modified-live virus vaccine administered to calves with maternal antibodies and challenged seven months later with a virulent bovine viral diarrhea type 2 virus. The Bovine Practitioner 43:35-43.<br /> <br /> Kuckleburg, C. J., D. J. McClenahan, and C. J. Czuprynski. 2008. Platelet activation by Histophilus somni and its LOS induces endothelial cell pro-inflammatory responses and platelet internalization. Shock 29:189-196.<br /> <br /> Kuckleburg CJ, Tiwari R, Czuprynski CJ. 2008. Endothelial cell apoptosis induced by bacteria-activated platelets requires caspase-8 and -9 and generation of reactive oxygen species. Thromb. Haemost. 99:363-372.<br /> <br /> McClenahan, D., K. Hellenbrand, D. Atapattu, N. Aulik, D. Carlton, A. Kapur, and C.J. Czuprynski. 2008. Effects of lipopolysaccharide and Mannheimia haemolytica leukotoxin on bovine lung microvascular endothelial cells and alveolar epithelial cells. Clin. Vaccine Immunol. 15:338-347.<br /> <br /> Atapattu DN, Albrecht RM, McClenahan DJ, Czuprynski CJ. 2008. Dynamin-2-dependent targeting of Mannheimia haemolytica leukotoxin to mitochondrial cyclophilin D in bovine lymphoblastoid cells. Infect Immun. 76:5357-5365.<br /> <br /> Kisiela DI, Czuprynski CJ.2009. Identification of Mannheimia haemolytica adhesins involved in binding to bovine bronchial epithelial cells. Infect Immun. 2009 77:446-455.<br /> <br /> McClenahan D, Hillenbrand K, Kapur A, Carlton D, Czuprynski C. 2009. Effects of extracellular ATP on bovine lung endothelial and epithelial cell monolayer morphologies, apoptoses, and permeabilities. Clin Vaccine Immunol. 16:43-48.<br /> <br /> Atapattu DN, Aulik NA, McCaslin DR, Czuprynski CJ. 2009. Brief heat treatment increases cytotoxicity of Mannheimia haemolytica leukotoxin in an LFA-1 independent manner. Microb Pathog. 46:159-165.<br /> <br /> Rivera-Rivas JJ, Kisiela D, Czuprynski CJ. 2009. Bovine herpesvirus type 1 infection of bovine bronchial epithelial cells increases neutrophil adhesion and activation. Vet Immunol Immunopathol. Apr 11. [Epub ahead of print].<br /> <br /> Tiwari R, Sullivan J, Czuprynski CJ. 2009. PECAM-1 is involved in neutrophil transmigration across Histophilus somni treated bovine brain endothelial cells. Microb Pathog. Jun 12. [Epub ahead of print]<br /> <br /> Presentations<br /> <br /> Woolums AR and Chase CCL. New discoveries in cattle health: how your actions influence funding decisions. Proceedings of the 41st Meeting of the American Association of Bovine Practitioners. September 25-27, 2008. Charlotte, NC, p. 205-208.<br /> <br /> Krunkosky TM, Jarrett CL, Ard MB, Bettis CL, Berghaus LJ, Hurley KAE, Woolums AR. Establishment of Bovine Primary Respiratory Epithelial Cell Cultures From Live Calves and Evaluation of Cell Responses to Bovine Respiratory Syncytial Virus Infection. Proceedings of the 89th Annual Meeting of the Conference for Research Workers in Animal Disease. December 7-9, 2008. Chicago, IL, p. 155.<br /> <br /> Woolums AR. Practical approaches to vaccination of the calfvaccinating in the face of maternal antibody. 15th American Dairy Science Association Discover Conference on Food Animal Agriculture: Biology of the Calf, Birth to 4 Months. Nov. 16-19, 2008. Roanoke, VA.<br /> <br /> Woolums AR. The bovine immune response to BRSV infection: helpful or harmful? Western Canadian Association of Bovine Practitioners 18th Annual Conference. January 15-17, 2009. Saskatoon SK, Canada.<br /> <br /> Woolums AR. Effectively vaccinating calves to prevent respiratory disease. Western Canadian Association of Bovine Practitioners 18th Annual Conference. January 15-17, 2009. Saskatoon SK, Canada.<br /> <br /> Woolums AR. Feedlot AIP: what the heck causes it? Western Canadian Association of Bovine Practitioners 18th Annual Conference. January 15-17, 2009. Saskatoon SK, Canada.<br /> <br /> Woolums AR, Krunkosky TM, Jarrett CL, Ard MB, Berghaus LJ, Bettis CL, Hurley KAE. Development of primary bovine bronchial epithelial cell (BBEC) cultures from live calves, and evaluation of cell responses to viral infection. 2009 American College of Veterinary Internal Medicine Annual Forum, June 3-6, 2009, Montreal QC, Canada. <br /> <br /> Berghaus LJ, Blas-Machado U, Vandenplas ML, Galland KL, Davis CE, Hurley KAE, Woolums AR. Comparison of three methods of identification of nasal BRSV shedding in calves. 5th International Veterinary Vaccines and Diagnostics Conference, July 19-23, Madison, WI, P12.<br /> <br /> Woolums A, Krunkosky T, Jarrett C, Ard M, Bettis C, Berghaus L, Hurley K, Kyser J. Establishment of bovine primary bronchial epithelial cell cultures from live calves and evaluation of cell responses to bovine respiratory syncytial virus (BRSV) infection. 2009 Bovine Respiratory Disease Symposium, August 5-6, 2009, Colorado Springs, CO, p. 82. <br /> <br /> Hurley DJ. Role of maternally derived white blood cells passed in the colostrum on immune function. 15th American Dairy Science Association Discover Conference on Food Animal Agriculture: Biology of the Calf, Birth to 4 Months. Nov. 16-19, 2008. Roanoke, VA.<br /> <br /> Hurley DJ. The transfer of maternal immunity and its impact on calf health and productivity. Academy of Veterinary Consultants Summer Meeting, August 6-8, 2009.<br /> <br /> Hurley DJ. An overview of the host immune response to bacterial and viral pathogens. Academy of Veterinary Consultants Summer Meeting, August 6-8, 2009.<br /> <br /> Rosenbusch, R. F., and N. Lee. Polyamine transport as virulence factor in Mycoplasma bovis: role in resistance to attack by macrophages. Poster G-007, 109th General Meeting of the American Society for Microbiology, Philadelphia, PA, 2009.<br /> <br /> Rosenbusch, R.F., N. Lee, and L. Christensen. Identification of Mycoplasma<br /> bovis genes involved in uptake of unopsonized mycoplasmas by activated <br /> macrophages. 17th International Organization for Microbiology Congress, Poster<br /> 133, Tianjin, China, 2008.<br /> <br /> Rosenbusch, R.F., N. Lee, D. R. Zamzow, C. D. Riedell, and L. Christensen. Use of a respiratory tract infection model in cattle to screen transposon 4001 mutants of Mycoplasma bovis. 17th International Organization for Microbiology Congress, Presentation in Seminar IV: Molecular and cell biology of mollicutes, Tianjin, China, 2008.<br /> <br /> Lee, N., D. R. Zamzow, C. D. Riedell, and R. F. Rosenbusch. Development of a transposon 4001 mutant library of a pathogenic Mycoplasma bovis strain. 17th International Organization for Microbiology Congress, Poster 207, Tianjin, China, 2008.<br /> <br /> Fulton, R.W., Blood, K.S., Panciera, R.J., Payton, M. Ridpath, J.F., Saliki, J.T., Burge, L.J., Confer, A.W., Welsh, R.D., Johnson, B.J., Reck.: Lung Pathology and Infectious Agents in Fatal Feedlot Pneumonias and Relationship with Animal and Treatment Information. American Association of Bovine Practitioners Proceedings. September 25-27, 2008. Charlotte, NC.<br /> <br /> Taylor, J.D., Fulton, R.W., Lehenbauer, T.W., Dabo, S.M., Confer, A.W.: Molecular Characterization of Pasteurella multocida Isolates from Fatal Feedlot Cases of Bovine Respiratory Disease. American Association of Bovine Practitioners Proceedings. September 25-27, 2008. Charlotte, NC.<br /> <br /> Ridpath, J.F., Vander Lay, B.L., Sweiger, S.H., Fulton, R.W., Kirkland, P.D.: Prevalence and Antigenic Differences of Bovine Viral Diarrhea Virus Subgenotypes Isolated from Cattle in Australia and the U.S. 51th Annual Meeting of AAVLD, October 22-27, 2008. Greensboro, NC.<br /> <br /> Fulton, R.W., Eberle, R., Burge, L.J., Reck, A.: Development and Use of Indirect ELISA Tests Detecting Antibodies to Bovine Coronavirus and Bovine Viral Diarrhea Virus in Acute and Convalescent Serums of Cattle. 51th Annual Meeting of AAVLD, October 22-27, 2008. Greensboro, NC.<br /> <br /> Shelton, T., Hoffman., Fulton.R.W., Ridpath, J.F.: Prevalence of Persistently Infected BVDV Dairy Calves from Dairies with BVDV Vaccination Programs and Distribution of BVDV Subtypes. 4th US BVDV Symposium, January 25-27, 2009. Phoenix, AZ.<br /> <br /> Fulton, R.W., Ridpath, J.F., Hessman, B.E., Blood, K.S.: Bovine Viral Diarrhea Virus Variability and Prevalence of BVDV Subtypes in Persistently Infected Cattle Entering Feedlots: BVDV1b as Predominant Subtype. 4th US BVDV Symposium, January 25-27, 2009. Phoenix, AZ.<br /> <br /> Ridpath, J.F., Neil, J., Fulton, R.W., Kirkland, P.D.: Bovine Viral Diarrhea Virus Type Subgenotypes Isolated from Cattle in the U.S. and Australia: Prevalence and Antigenic Differences. 4th US BVDV Symposium, January 25-27, 2009. Phoenix, AZ.<br /> <br /> Fulton, R.W.: BVD Subtypes: What Matters in the Real World. Western Veterinary Conference. February 17, 2009. Las Vegas, NV.<br /> <br /> Confer AW, Ayalew S, Montelongo M, Step DL, Wray JH, Hansen RD, Panciera RJ. Immunity of cattle following vaccination of calves with a Mannheimia haemolytica Chimeric PlpE-LKT (SAC89) protein. International Society of Pasteurellaceae Conference, Sorrento, Italy, October 2008. P403<br /> Ayalew S, Confer AW. Immunoproteomic approach to identify and characterize immunologically important Mannheimia haemolytica outer membrane proteins. International Society of Pasteurellaceae Conference, Sorrento, Italy, October 2008. P402<br /> Confer AW. Comparative Pathology of and Pathogenic Mechanisms in Bovine Bacterial Pneumonia. Proceedings, ACVIM, Montreal, June 2009<br /> Confer AW. Immunity against Mannheimia haemolytica: Current knowledge and New Approaches. Proceedings, ACVIM, Montreal, June 2009<br /> Chase CCL, LJ Braun, L Holler, M Spire. Comparing cytopathic and noncytopathic bovine viral diarrhea virus (BVDV) vaccines: antigen trafficking increased to mucosal surfaces with NCP vaccines. 41st annual conference of the American Association of Bovine Practitioners, Charlotte, NC, September 25-27, 2008.<br /> <br /> Chase CCL, LJ Braun, J Ridpath, R Harland. Comparison of the immune response between a pair of noncytopathic and cytopathic bovine viral diarrhea virus (BVDV) type 1 isolates. 41st annual conference of the American Association of Bovine Practitioners, Charlotte, NC, September 25-27, 2008.<br /> <br /> Chase CCL, A Young, A Zimmerman, K Gilkerson, K Barling, D Scholz. The induction of a T helper 1 immune response with an inactivated viral vaccine. . 41st annual conference of the American Association of Bovine Practitioners, Charlotte, NC, September 25-27, 2008.<br /> <br /> Chase C. Bovine viral diarrhea virus persistent infection of white-tailed deer and their risk to domestic cattle. CSREES Awardee Workshop, Chicago, IL, December 7, 2008.<br /> Zimmerman AD, RE Buterbaugh, JA Schnakel, CCL Chase. 4th US BVDV Symposium, January 26, 2009, Phoenix AZ.<br /> <br /> Chase CCL, LJ Braun, L Holler, M Spire. Comparing CP and NCP BVDV vaccines: antigen trafficking increased to mucosal surfaces with NCP vaccines. 4th US BVDV Symposium, January 26, 2009, Phoenix AZ.<br /> <br /> Chase CCL, A Young, A Zimmerman, K Gilkerson, K Barling, D Scholz. The induction of a T helper 1 immune response with an inactivated viral vaccine. 4th US BVDV Symposium, January 26, 2009, Phoenix AZ.<br /> <br /> Chase CCL, LJ Braun, J Ridpath, R Harland. Comparison of the immune response between a pair of NCP and CP bovine viral diarrhea (BVDV) type 1 isolates. 4th US BVDV Symposium, January 26, 2009, Phoenix AZ.<br />

Impact Statements

1. Objective 1: Respiratory epithelial cell research can provide researchers with tools to uncover complexities of polymicrobial infections without use of expensive animals. The studies of behavioral and physiologic parameters of cattle with BRD will uncover the relationship between clinical and behavioral parameters and morphologic evidence of pneumonia. Field studies of viral etiologies interacting with M. haemolytica and P. multocida will impact understanding. Examination of antigenic diversity of BVDV will be relevant to current and future vaccines.
2. Objective 2: The impact of the studies of intranasal vaccination IFOMA is that they could provide the basis for new management practices that improve the ability of calves to respond to vaccines to prevent respiratory disease. This would lead to decreased financial loss and increased profits for cattle producers, who would have to spend less money treating respiratory disease in calves, and would realize greater profits from healthier, more productive cattle.
3. Objective 3: If siRNA can suppress BRSV replication, it could be used as an antiviral therapy to specifically protect cattle from infection with BRSF or other viruses. Two virulence genes of Mycoplasma bovis were identified that may be target genes to engineer specific gene deletions in M. bovis that can be used as modified-live vaccines against M. bovis-associated respiratory diseases of cattle. Improving handling and nutrition in the feedlot will have a positive effect on BRDC.

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Report Information

Participants

See meeting minutes for list of participants.

Brief Summary of Minutes

See attached meeting minutes.

Accomplishments

Objective 1: To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents.<br /> <br /> Short term outcomes: Commercially available assays for human respiratory syncytial virus (RSV) were shown to be effective for use in identification of bovine RSV (BRSV). The infectious agents associated with fatal bovine respiratory disease (BRD) in feedlots were characterized, providing a much-needed updating of our understanding of the pathogens involved in fatal feedlot BRD, and the relationship between specific infectious agents and specific pathologic lesions. The prevalence of different genotypes of bovine viral diarrhea virus (BVDV) in U.S. and Australian diagnostic lab isolates was determined; it was found that there is a difference in the genotypes most prevalent in the two countries. Annual summaries of the number of major pathogens isolated from BRD cases presented to regional diagnostic laboratories were disseminated to the membership. <br /> <br /> Outputs: An annual summary of the frequency of isolation of major pathogens from BRD cases presented to regional diagnostic laboratories was made available to the membership. Research abstracts were presented that demonstrated that a commercial assay for human RSV can be used to identify BRSV and which described the prevalence of various genotypes of BVDV. Research papers were published which described the prevalence of various BVDV genotypes in U.S. cattle populations. <br /> <br /> <br /> Activities: Project members worked together to share prevalence data on pathogens isolated in BRD cases presented to regional diagnostic laboratories, and to characterize the distribution of BVDV genotypes in various U.S. cattle populations. <br /> <br /> <br /> Objective 2: To investigate the basic biology, molecular pathogenesis, and<br /> immunopathogenesis of polymicrobial infections including important viral and bacterial agents.<br /> <br /> Short term outcomes: An in vitro system was developed which allows assessment of the interaction of infected bovine respiratory epithelial with cells of the immune response, providing a method of studying airway epithelial cells, which because of restrictions of anatomy and physiology cannot be studied in the living animal. Immune cells from cattle infected with Mannheimia haemolytica (M. haemolytica) were shown to produce increased amounts of substance P, which likely contributes to increased vascular leakage and edema in the lungs of infected cattle. Proteogenomic mapping was developed to better characterize the relationship between genes, the proteins they encode, and the function of those proteins for M. haemolytica. Proteomic analysis was used to evaluate the mechanism by which BVDV interferes with function of dendritic cells, which initiate the immune response to the virus; acute phase protein signaling was most commonly modified. A molecule which may be among the most important proteins regulating bovine herpesvirus-1 (BHV-1) replication was evaluated extensively and was shown to interfere with host immune function; the means by which this molecule interacts with host cell proteins to enhance BHV-1 replication was also deduced. The methods by which BHV-1 establishes latency and reactivates from latency (which is among the most potent of the virus disease inducing strategies) were characterized extensively. Methods of typing Pasteurella multocida (P. multocida) strains in order to distinguish between commensal and virulent isolates were developed. A model of BVDV and M. haemolytica co-infection utilizing exposure to naturally PI cattle was validated, and co-infection was shown to impact metabolism and productivity of feedlot cattle. A method of evaluating the effect of BVDV on development of immunity in the fetal calf was developed which utilizes a strategy to isolate immune cells from the fetal liver. The impact of BVDV on the cellular structure of macrophages, a critical immune cell, was determined. Research was undertaken which demonstrated the interaction of M. haemolytica leukotoxin with mitochondria, revealing a new pathogenic mechanism by which the leukotoxin causes disease. The role of neutrophil DNA traps (or nets) in the control of M. haemolytica was determined. <br /> <br /> Outputs: Presentations were made to veterinarians at annual veterinary continuing education conferences on the latest information regarding BRSV, Mycoplasma bovis (M. bovis), and on vaccination to control respiratory disease in cattle. Research abstracts were presented that demonstrated the interaction between infected bovine respiratory epithelial cells and immune cells, which demonstrated how molecular transport systems are related to pathogenicity of M. bovis; which demonstrated the relationship between quantitative and qualitative load of bacterial pathogens and respiratory disease; that provided information on proteogenomic mapping of the M. haemolytica genome; that showed that cytopathic BVDV and noncytopathic BVDV differentially modulate induction of the host immune response; that described the effect of BHV-1 infection of airway epithelial cells to increase activation of the host immune response; and that described the region of the M. haemolytica leukotoxin which specifically interacts with mitochondria. <br /> Research papers were published which demonstrated the effect of M. bovis to suppress leukocyte antibacterial responses; which demonstrated the effect of M. bovis on growth and health in feedlot cattle, which demonstrated that BVDV interferes with the initiation of the host immune response by impairing the function of professional antigen presenting cells; which showed how a protein encoded by BHV-1 impairs the host interferon response and thus counteracts host immunity; which demonstrated how BHV-1 undergoes latency and reactivates from latency; which reported the current relationship between infectious agents and various pathologic agents in feedlot cattle; which reported the knowledge gaps impacting the development of BVDV control programs; which outlined methods for identifying pathogenic vs. commensal P. multocida isolates; which outlined the current evidence regarding risk factors and effective control measures in BRD; and which presented the results of immunoproteomic analysis of M. haemolytica outer membrane proteins.<br /> <br /> Activities: Project members worked to test the effect of various vaccination strategies on productivity and health of feedlot cattle, and to disseminate this information to veterinarians and producers. Work on the Genetics of Feedlot Health Project was continued, which evaluated the role of genetics, nutrition, behavior, and infection and immunity on the occurrence of feedlot respiratory disease and carcass quality. <br /> <br /> <br /> Objective 3: To develop management and prevention strategies that incorporate<br /> new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss.<br /> <br /> Short term outcomes: A research trial was completed which evaluated the response of young calves to intranasal or subcutaneous vaccination to improve immunity to respiratory infections; the data indicated that one commercial vaccine induced better immunity when given subcutaneously. A gene deleted M. bovis vaccine was developed which could be given intranasally and did not disseminate to other sites in the body, indicating that it was stable and safe. Seroconversion to M. bovis was found to be associated with decreased growth of feedlot cattle. It was found that subjective and laboratory indicators of disease were not good measures of disease progression, but measures of activity (by pedometers) showed promise in providing accurate assessment of disease progression. A research trial was carried out which confirmed that vaccination of calves before weaning improved their health and productivity when they were exposed to cattle persistently infected (PI) with BVDV in the feedlot. Immunoproteomic analysis was used to identify outer membrane proteins of M. haemolytica which should form the basis for improved vaccines. A research trial was completed which showed that vaccination once was as effective as vaccination twice to prevent respiratory disease in feedlot cattle; however, cattle vaccinated twice had improved feed efficiency. <br /> <br /> Outputs: Presentations were made to veterinarians and producers at continuing education meetings on the accurate prediction of respiratory disease risk based on daily morbidity and mortality counts, and on the approach to eradicating BVDV in a region (Upper Peninsula of Michigan). Research abstracts were presented that compared the results of vaccinating calves intranasally or subcutaneously to improve immunity to respiratory infection; that demonstrated how genetic modifications can be applied to the development of M. bovis vaccines; that demonstrated that vaccination mitigates the adverse health effects of exposure to BVDV PIs in feedlots; that demonstrated the feasibility of BVDV eradication on a regional level; that described the degree to which whitetail deer infected with BVDV can spread disease to cattle; and that described a method for isolating fetal bovine immune cells from the liver. Research papers were published that presented the impact of various vaccine-parasiticide combinations on health and productivity in beef cattle, that demonstrated the use of rumen temperature reporting boluses to monitor fever in cattle co-infected with BVDV and M. haemolytica; and which demonstrated the efficacy of a new intranasal M. haemolytica vaccine containing choleratoxin as an adjuvant. <br /> <br /> Activities: Project members worked to assess how vaccination strategies and evaluation of physiologic and behavioral indicators could be used to improve health and productivity of cattle in field settings. A project to eradicate BVDV in the UP of MI continued to progress; producers were engaged to work with project researchers to identify and remove persistently infected cattle and to carry out surveillance and management practices to advance the project.<br /> <br /> <br /> Plans for the coming year: This is the final year of the current project; major plans are to continue to undertake surveillance of the rate of isolation of major BRD pathogens by regional diagnostic laboratories, to cooperate in research that provides foundational knowledge regarding the basic mechanisms by which infectious BRD agents interact with the bovine immune response, and to test and disseminate to veterinarians and producers practices that can be translated to the field to decrease the impact of BRD in U.S. cattle. <br /> <br /> NC-1027 will also hold its annual meeting in conjunction with the American Association of Bovine Practitioners (AABP) Convention in St. Louis on September 21-22. NC-1027 has met in conjunction with AABP for several years, and this has provided an excellent opportunity for the Project to inform veterinarians of new developments in the science of BRD control, and to learn from veterinarians the current problems on which research should be focused. In 2010 NC-1027 met in conjunction with AFRI awardees conducting research on BRD; this also provided a new opportunity for improved collaboration between scientists working on BRD from different perspectives. In 2011 NC-1027 is again planning to meet in conjunction with AFRI awardees and to offer the meeting as a pre-conference seminar billed as Cutting Edge Science Related to BRD. This will allow veterinarians and other AABP attendees the opportunity to hear the latest developments in BRD prevention and control, and will allow NC-1027 members and AFRI awardees the opportunity to discuss questions and receive input from veterinarians working in the field.<br /> <br /> Members of NC-1027 have also been in contact with the investigators leading the new BRD CAP; plans are underway to coordinate efforts between NC-1027 and the CAP to develop impactful outreach efforts that will deliver information from CAP activities to veterinarians, scientists in industry and academia, and producers. Cooperation between NC-1027 and the CAP should lead to a high level of impact, as the CAP scientists will benefit from the historical perspective and experience of NC-1027, and the established links NC-1027 has developed with AABP, the Academy of Veterinary Consultants, the National Cattlemens Beef Association, and the American Association of Veterinary Laboratory Diagnosticians. <br /> <br /> <br />

Publications

Ammari, M. G., McCarthy, F. M., Nanduri, B., Pinchuk, L.M. Analysis of Bovine Viral Diarrhea Viruses-infected monocytes: identification of cytopathic and non-cytopathic biotype differences. BMC Bioinformatics. 11 SUPPL, 2010, MCBIOS, in press.<br /> <br /> Atapattu DN, Aulik NA, McCaslin DR, Czuprynski CJ. 2009. Brief heat treatment increases cytotoxicity of Mannheimia haemolytica leukotoxin in an LFA-1 independent manner. Microb Pathog. 46:159-165.<br /> <br /> <br /> Ayalew S, Step DL, Montelongo M, Confer AW. Intranasal vaccination of calves with Mannheimia haemolytica chimeric protein containing the major surface epitope of outer membrane lipoprotein PlpE, the neutralizing epitope of leukotoxin, and cholera toxin subunit B. Vet Immun & Immunopathol 132: 295-302, 2009<br /> <br /> Ayalew S, Confer AW, Hartson SD, Shrestha B. Immunoproteomic Analysis of Outer<br /> Membrane Proteins of Mannheimia haemolytica and Identification of Potential Vaccine Candidates. Proteomics 10: 2151-2164, 2010<br /> <br /> Brum M, Coats C., Sangena B.R, Doster.A., Jones, C. and Chowdhury.S.I. (2009). Role of Envelope proteins gE in the anterograde transport of BHV-1 following reactivation in the Trigeminal Ganglia. J. Neuro virology. 15 (2): 196-201.<br /> <br /> Brum. M.C.S., C. Coats, B.R. Sangena, A. Doster, C. Jones, and S.I. Chowdhury.<br /> 2009. Bovine herpesvirus type 1 (BoHV-1) anterograde neuronal transport from<br /> trigeminal ganglia to nose and eye requires glycoprotein E. J. Neurovirology, 15:1-6.<br /> <br /> Burciaga-Robles, L.O., Step, D.L., Krehbiel, C.R., Holland, B.P., Richards, C.J., Montelongo, M.A., Confer, A.W., Fulton, R.W.: Exposure by Persistently Infected Calves with Bovine Viral Diarrhea Virus Type 1b and Subsequent Infection with Mannheimia haemolytica Demonstrating Effects on Clinical Signs and Immune Parameters: Model for Bovine Respiratory Disease Via Viral and Bacterial Interaction. Journal of Animal Science.88: 2166-2178, 2010.<br /> <br /> Burciaga-Robles, L.O., Krehbiel, C.R., Step, D.L., Holland, B. P., Richards, C.J., Montelongo, M.A., Confer, A.W., Fulton, R.W.: Effects of Exposure to Calves Persistently Infected with Bovine Viral Diarrhea Virus Type 1b and Mannheimia haemolytica Challenge on Animal Performance, N Balance, and Visceral Organ Mass in Beef Steers. Journal of Animal Science. 88:2179-2188, 2010.<br /> <br /> Burge, L.J., Welsh, R.D., Johnson, B.J., Reck. A.: Lung Pathology and Infectious Agents in Fatal Feedlot Pneumonias and Relationship with Mortality, Disease Onset, and Treatments. Journal of Veterinary Diagnostic Investigation 21: 464-477, 2009.<br /> <br /> Chowdhury, S. and C. Jones. Bovine herpesvirus type 1 (BHV-1) is an important<br /> cofactor in the bovine respiratory disease complex. Veterinary Clinics of North America: IN PRESS, Food Animal Practice.<br /> <br /> Confer AW. Update on bacterial pathogenesis in BRD. Animal Health Res Rev 10: 145-149: 2009.<br /> <br /> Confer AW, Ayalew S, Step D L, Trojan B, Montelongo M. Intranasal vaccination of young Holstein calves with Mannheimia haemolytica chimeric protein PlpE-LKT (SAC89) and cholera toxin Vet Immun & Immunopathol 132: 232-236, 2009<br /> <br /> Czuprynski, C.J. 2009. Host response to bovine respiratory pathogens. Anim Health Res Rev. 2009 10:141-143.<br /> <br /> Ellis, J., S. Gow, N. Goji, C. Jones, A. Workman, G. Henderson, G. Alaniz, and T. Meinert. 2009. Efficacy of a combination viral vaccine in protecting cattle from experimental infection with bovine herpesviruses-1 isolated from recent vaccine breaks. J of American Veterinary Medical Association, 235:563-572.<br /> <br /> Fulton, R.W., Whitley, E.M., Johnson, B.J., Ridpath, J.F., Kapil, S., Burge, L.J., Cook, B.J., Conf er, A.W.: Prevalence of Bovine Viral Diarrhea Virus (BVDV) in Persistently Infected Cattle and BVDV Subtypes in Affected Cattle in Beef Herds in South Central United States. Canadian Journal for Veterinary Research 73: 283-291, 2009.<br /> <br /> Fulton, R.W.: Bovine Respiratory Disease Research: 1983-2009. Animal Health Research Reviews. 10: 131- 140, 2009.<br /> <br /> Hanzlicek G, White B, Anderson D, Mosier D, Renter D. Pathological and<br /> physiological changes following induced Mannheimia pneumonia in beef feeder calves. Amer J Vet Res. Mar 2010. 71(3):359-369.<br /> <br /> Hanzlicek G, White B, Renter D, Blasi D. A field study evaluating health,<br /> performance, and behavior differences in crossbred beef calves administered different vaccine-parasiticide product combinations. Vaccine. In press.<br /> <br /> Henningson, J.L., C.L.Topliff, L.H.Gil, R.O.Donis, D.J.Steffen, K.M.Eskridge, C.L.Kelling. 2009. Influence of Npro on BVDV virulence and interferon type induction in calves. American Journal of Veterinary Research 70: 1117-1123.<br /> <br /> Jaber, T., A. Workman, and C. Jones. Small non-coding RNAs encoded within<br /> the bovine herpesvirus 1 latency related gene can reduce steady state levels of<br /> infected cell protein 0 (bICP0). 2010. J Virology, 84: 62976307<br /> <br /> Jones, C. 2009. Regulation of Innate Immune Responses by Bovine Herpesvirus 1<br /> and Infected Cell Protein 0 (bICP0). Viruses 1:255-275.<br /> <br /> Jones C, Chowdhury S (2010). Bovine herpesvirus type 1 (BHV-1) is an important cofactor in the bovine respiratory disease complex. Vet. Clin. Food Anim. Practice. (In Press).<br /> <br /> Kisiela DI, Czuprynski CJ.2009. Identification of Mannheimia haemolytica adhesins involved in binding to bovine bronchial epithelial cells. Infect Immun. 2009 77:446-455.<br /> <br /> Kisiela DI, Aulik NA, Atapattu DN, Czuprynski ,CJ. 2010. N-terminal region of<br /> Mannheimia haemolytica leukotoxin serves as a mitochondrial targeting signal in<br /> mammalian cells. Cell. Microbiol. 12:976-987.<br /> <br /> Lee, S.-R., Nanduri, B., Pharr, G. T., Stokes, J. V., Pinchuk, L. M. 2009. Bovine Viral Diarrhea Virus infection affects the expression of proteins related to professional antigen presentation in bovine monocytes. Biochim. Biophys. Acta. 1794:14-22.<br /> <br /> Leyh, R.D., Fulton, R.W., Stegner, J.E., Goodyear, M., Witte, S., Taylor, L.P., Johnson, B.J., Step, D.L., Ridpath, J.F., Holland, B.P.: Fetal Protection in Heifers Vaccinated with Modified Live Virus Vaccine Containing Bovine Viral Diarrhea Virus Subtypes BVDV1a and BVDV2a after Challenge with BVDV1b Persistently Infected Cattle. Accepted for publication, 2010. American Journal for Veterinary Research.<br /> <br /> Liang, D., L. Chen, I.H. Ansari, L. H. Gil, C.L. Topliff, C.L. Kelling, R.O.Donis. 2009. A replicon trans-packing system reveals the requirement of nonstructural proteins for the assembly of bovine viral diarrhea virus (BVDV) virion. Virology 387:331-340.<br /> <br /> McClenahan D, Hillenbrand K, Kapur A, Carlton D, Czuprynski C. 2009. Effects of<br /> extracellular ATP on bovine lung endothelial and epithelial cell monolayer morphologies, apoptoses, and permeabilities. Clin Vaccine Immunol. 16:43-48.<br /> <br /> Meyer, F. and C. Jones. 2009. C/EBP-alpha cooperates with bTIF to activate the<br /> bovine herpesvirus 1 immediate early transcription unit 1 promoter. J. Neurovirology 15:123-130.<br /> <br /> Panciera RJ, Confer AW. Pathogenesis and pathology of bovine pneumonia. Vet Clin North Amer Food Anim Prac 26: 191-214, 2010<br /> <br /> Ridpath, J.F., Fulton, R.W.: Knowledge Gaps Impacting the Development of Bovine Viral Diarrhea Virus Control Programs in the United States. Journal of American Veterinary Medical Association 235: 1171-1179, 2009.<br /> <br /> Ridpath, J.F., Fulton, R.W., Kirkland, P.D., Neil, J.: Prevalence and Antigenic Differences Observed Between Bovine Viral Diarrhea Virus Subgenotypes Isolated From Cattle in Australia and Feedlots in the Southwestern United States. Journal of Veterinary Diagnostic Investigation 22; 184-191, 2010.<br /> <br /> Rivera-Rivas JJ, Kisiela D, Czuprynski CJ. 2009. Bovine herpesvirus type 1 infection of bovine bronchial epithelial cells increases neutrophil adhesion and activation. Vet Immunol Immunopathol. 131:167-176.<br /> <br /> Rose-Dye, T.K., Burciaga-Robles, L.O., Krehbiel, C.R., Step, D.L., Fulton, R.W., Confer. A.W., Richards, C.J.: Rumen Temperature Change Monitored with Remote Reporting Rumen Temperature Boluses Following Challenge with Bovine Viral Diarrhea Virus and Mannheimia haemolytica. Journal of Animal Science, Accepted for publication, 2010.<br /> <br /> Rosenbusch R, Woolums A, Grooms D, Larson R. Summary of breakout sessions of 2009 Bovine Respiratory Disease Symposium. An Health Res Rev 2009;10:169-171.<br /> <br /> Saira, K., Y. Zhou, and C. Jones. 2009. The infected cell protein 0 encoded by bovine herpesvirus 1 (bICP0) associates with interferon regulatory factor 7 (IRF7), and consequently inhibits beta interferon promoter activity. J. Virology. 83:3977-3981.<br /> <br /> Shanthalingham, S., C.L.Topliff, C.L.Kelling, S.Srikumaran. 2010. Bighorn sheep fetal lung cell line for detection of respiratory viruses. Canadian Journal of Veterinary Research 74:75-77.<br /> <br /> Singh K, Ritchey JW, Confer AW. Mannheimia haemolytica: Bacterial-host interactions in bovine pneumonia. Vet Pathol 2010, in press<br /> <br /> Singh K, Confer AW, Hope JC, Rizzi T, Wyckoff JW, Weng HY, Ritchey JW. Cytotoxicity and cytokine production by bovine alveolar macrophages challenged with wild type and leukotoxindeficient Mannheimia haemolytica. The Vet J 2010, Epub ahead of print.<br /> <br /> Step, D.L., Krehbiel, C.R.., Burciaga-Robles, L.O., Holland, B.P., Fulton, R.W., Confer, A.W., Bechtol, D.T., Brister, D.L., Hutcheson, J.P., Newcomb, H.L.: Comparison of Single Vaccination Versus Revaccination with a Modified Live Virus Vaccine Containing Bovine Herpesvirus-1, Bovine Viral Diarrhea Virus (Types 1a and 2a), Parainfluenza Type 3 Virus, and Bovine Respiratory Syncytial Virus in the Prevention of Bovine Respiratory Disease. Journal of American Veterinary Medical Association 235: 580-587, 2009.<br /> <br /> Stevens ET, AD Zimmerman , RE Buterbaugh, K Barling, D Scholz, J Rhoades ,<br /> CCL Chase. 2009. The Induction of a Cell-Mediated Immune Response to<br /> Bovine Viral Diarrhea Virus with an Adjuvanted Inactivated Vaccine. Vet<br /> Therapeutics 10(4):E1-8.<br /> <br /> Taylor, J.D., Fulton, R.W., Dabo, S.M., Lehenbauer, T.W., Confer, A.W.: Comparison of Genotypic and Phenotypic Characterization Methods for Pasteurella multocida Isolates from Fatal Cases of Bovine Respiratory Disease. Journal of Veterinary Diagnostic Investigation 22; 366-375, 2010.<br /> <br /> Taylor, J.D., Fulton, R.W., Lehenbauer, T.W., Step, D.L., Confer, A.W.: The Epidemiology of Bovine Respiratory Disease: What is The Evidence for Predisposing Factors? In press, 2010. Canadian Veterinary Journal.<br /> <br /> Taylor, J.D., Fulton R.W., Lehenbauer T.W., Step D.L, Confer A.W.: The Epidemiology of Bovine Respiratory Disease: What is the Evidence for Preventive Measures? In press, 2010. Canadian Veterinary Journal.<br /> <br /> Tiwari R, Sullivan J, Czuprynski CJ. 2009. PECAM-1 is involved in neutrophil transmigration across Histophilus somni treated bovine brain endothelial cells. Microb Pathog. 47:164-170.<br /> <br /> Topliff C.L., D.R. Smith, S.L.Clowser,D.J. Steffen, J.N. Henningson, B.W. Brodersen, D. Bedenice, R. J. Callan, C.Reggiardo, K.L. Kurth, C.L. Kelling. 2009. Prevalence of bovine viral diarrhea virus infections in alpacas in the United States. Journal of American Veterinary Medical Association 234:519-529.<br /> <br /> <br /> White B, Hanzlicek G, Sanderson M, Anderson D, Larson R. Mollicutes<br /> species and Mycoplasma bovis prevalence and association with health outcomes in beef feeder calves at arrival and initial treatment for bovine respiratory disease. Can Vet J. 2010;51: In Press.<br /> <br /> Wiggins MC, Woolums AR, Hurley DJ, Sanchez S, Ensley DT, Donovan D. The effect of various Mycoplasma bovis isolates on bovine leukocyte responses. Comp Immunol Microbiol Infect Dis Epub ahead of print. 2010. doi:10.1016/j.cimid.2010.02.001<br /> <br /> Woolums A, Chase C, Fulton R, Rosenbusch R, Tremblay R. Introduction to the Proceedings of the 2009 Bovine Respiratory Disease Symposium. An Health Res Rev 2009;10:99.<br /> <br /> Workman, A., S. Perez, A. Doster, and C. Jones. 2009. Dexamethasone treatment<br /> of calves latently infected with bovine herpesvirus 1 (BHV-1) leads to activation of the bICP0 early promoter, in part by the cellular transcription factor C/EBP-alpha. J. Virology, 83:8800-8809.<br /> <br /> Workman, A. and C. Jones. Bovine herpesvirus 1 (BoHV1) productive infection and bICP0 early promoter activity are stimulated by E2F1. 2010. J Virology, 84: 63086317<br /> <br /> <br />

Impact Statements

1. Ongoing work by the Committee to track the rate of isolation of major pathogens from BRD cases presented to regional diagnostic laboratories provides year-to-year surveillance on changes in the impact of various agents, and on the development of new or emerging pathogens. For example, increased surveillance of respiratory coronavirus which has occurred in recent years will allow scientists to determine the relative importance of this agent as an emerging cause of BRD.
2. Characterization of the prevalence of different BVDV genotypes in U.S. cattle populations will allow scientists to determine the degree to which BVDV is evolving over time, and to determine whether current vaccines are formulated with appropriate genotypes to optimally control disease in U.S. cattle.
3. The validation of a patient-side test for human RSV in the diagnosis of BRSV provides the basis for more rapid diagnosis of respiratory pathogens; this effort represents a direct response to stakeholders, who have repeatedly expressed a need for more rapid tests to diagnose BRD.
4. The in vitro system developed to allow assessment of the interaction of infected bovine respiratory epithelial with cells of the immune response provides a method of studying the responses of living airway epithelial cells, which because of restrictions of anatomy and physiology cannot be studied in the living animal
5. The effort to develop proteogenomic mapping of M. haemolytica will provide a valuable new tool for researchers attempting to better understand how the genome of a major pathogen translates to its disease-causing phenotype.
6. The proteomic analysis of immune cells infected with either cytopathic or noncytopathic BVDV provides foundational knowledge regarding a major bovine pathogen. The data obtained will thus provide the basis for future studies to determine targets for vaccines and therapeutics to minimize the negative effects of BVDV on bovine health and productivity.
7. The research to determine the molecular mechanisms by which BHV-1 impairs the host response and undergo latency are providing important foundational knowledge that is improving the ability of scientists to elucidate basic functions of an important viral pathogen. The information on latency will help improve understanding not only of BHV-1 but also of other herpesviral diseases which impair the health and productivity of agricultural animals, because all herpesviruses undergo latency and latency is currently poorly understood.
8. The new model of experimental BRD which utilizes field exposure to naturally BVDV PI cattle followed by M. haemolytica challenge will provide a real-life model for testing vaccines and management strategies designed to minimize the negative impacts of BVDV and BRD.
9. Work to determine the impact of BVDV on the development of immunity in the fetus will not only provide foundational knowledge regarding the interaction of infectious agents with the developing bovine immune system, but the model will also be useful for evaluating the pathogenesis of other diseases that impact bovine health and productivity by causing fetal disease and death
10. The research demonstrating that M. haemolytica leukotoxin causes cell damage in part through interaction with mitochondria provides foundational knowledge regarding a new mechanism of pathogenesis by a major BRD pathogen; it also provides a basis for the development of methods which could mitigate the damaging effect of leukotoxin on bovine immune cells.
11. The research describing the role of neutrophil traps (nets) in host resistance to M. haemolytica provides foundational knowledge regarding the immune response to bacterial pathogens; this could form the basis for future studies to develop therapies based on molecules which could bind to and inactivate M. haemolytica.
12. Early diagnosis of BRD remains a challenge for producers, particularly feedlot producers; thus, the research evaluating the use of clinical signs, laboratory tests, and measures of activity to identify BRD will provide valuable new information to veterinarians and producers which should improve early diagnosis of BRD and thus improve response to treatment.
13. The gene deleted strain developed by project researchers provides a new platform for the development of safe and effective vaccines for M. bovis.
14. The information gained from the trial evaluating the effect of respiratory vaccines given to young calves by either the intranasal or subcutaneous routes will help veterinarians make decisions about when to most effectively administer vaccines, and to choose the best route of administration.
15. The BVDV eradication project being undertaken in the UP of Michigan provides an invaluable test case on the feasibility of eradicating BVDV from the national herd, an effort which has been advocated by the Academy of Veterinary Consultants and other stakeholder groups. This project provides a unique opportunity to educate producers and to develop partnerships with various entities to meet a common goal of improving cattle health.
16. The research proving that the impact of exposure to BVDV can be mitigated by vaccination prior to feedlot entry provides producers with a valuable tool to counteract the impact of BVDV while it remains present in the U.S. herd.
17. The characterization, cloning, and production of subunit components of M. haemolytica and P. multocida offer opportunity for the development of improved vaccines based on the most current concepts in bacterial pathogenesis to control BRD.
18. Research testing the effect of single vaccination versus vaccination plus booster on health and performance of feedlot cattle will help veterinarians and producers decide whether two doses of vaccine are warranted in managing high risk cattle.
19. Improved understanding of the interplay of genetics, nutrition, and infection in feedlot respiratory disease will provide the possibility of selecting cattle resistant to BRD, or tailoring management strategies to the phenotype of different cattle groups to improve BRD control.

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Report Information

Participants

Attendees at NC-1027 Technical Committee meeting, Sept. 21, 2011
*voting member of participating station;

*Chase, Chris (christopher.chase@sdstate.edu), South Dakota State University;
*Czuprynski, Chuck (czuprync@svm.vetmed.wisc.edu), University of Wisconsin;
*Fulton, Robert (robert.fulton@okstate.edu), Oklahoma State University;
*Gershwin, Laurel (ljgershwin@ucdavis.edu), Universiy of California, Davis;
*Grooms, Dan (groomsd@cvm.msu.edu), Michigan State University;
*Woolums, Amelia (awoolums@uga.edu), University of Georgia;
Hamernik, Deb (dhamernik2@unl.edu), University of Nebraska, Lincoln (AA);
Renter, David (drenter@vet.ksu.edu), Kansas State University;
White, Brad (bwhite@vet.ksu.edu), Kansas State University;
Daly, Russ (russell.daly@sdstate.edu), South Dakota State University;
Davis, Jessica (jhd1985@gmail.com), VMTRC, UC Davis, Tulare;
Lehenbauer, Terry (tlehenbauer@ucdavis.edu), VMTRC, UC Davis, Tulare;
Tucker, Cassandra (cbtucker@ucdavis.edu), UC Davis;
Van Eenennaam, Alison (avaneenennaam@ucdavis.edu), UC Davis;
Holland, Margo (mholland@nifa.usda.gov), USDA-NIFA;
Johnson, Peter (pjohnson@nifa.usda.gov), USDA-NIFA;
Womack, James (jwomack@cvm.tamu.edu), VTPB, CVM Texas A&M;
Elliott, Jan (jselliott@cvm.tamu.edu), VTPB, CVM, Texas A&M;
Neibergs, Holly (neibergs@wsu.edu), Washington State University;

Brief Summary of Minutes

NC-1027 Annual Report, 2011
September 21, 2011
St. Louis, MO
In conjunction with the AABP Annual Convention


Joint Meeting of Multistate Research Committee NC 1027, An Integrated Approach To Control Of Bovine Respiratory Diseases (http://nimss.umd.edu/homepages/home.cfm?trackID=7796) and the USDA-NIFA BRD CAP

Objectives:
(1) Strengthen integration/coordination of USDA-NIFA competitive and multi-state formula-based projects on bovine respiratory disease
(2) Conduct NC 1027 (NC1092) business meeting
(3) Discuss future options to maintain maximum viability of a BRD multistate research committee

Agenda:

8:30 AM: Welcome
8:40-10:00 AM: BRDC CAP Presentations
8:40 - 9 General overview of the BRD CAP - James Womack
9-10 BRD CAP Ongoing research projects reports -
9:00-9:20 Terry Lehenbauer and Jessica Davis - Tulare Calf Trial
9:20-9:40 Laurel Gershwin - Challenge Trial
9:40-10:00 CassandraTucker - Behavior/Meloxicam Trial

10-10:30: Break
10:30-12 BRDC CAP Presentations and Discussion
10:30-11:00 Educational component of CAP- Alison Van Eenennaam
11:00-11:30 Outreach component of CAP  Alison Van Eenennaam
11:30-12:00 Collaborative opportunities for CAP Outreach with NC1027 (and NC-1192, the next regional project beginning in 2012)

12  1: Lunch, sponsored by Pfizer. Discussion of efforts to lobby for more animal health research funding as the 2012 Farm Bill is planned, led by Amelia Woolums.
1:00 - 2:00 PM: Updates from USDA-Agricultural Research Service (ARS) & USDA- National Institute of Food and Agriculture (NIFA) (Margo Holland and Peter Johnson, NIFA)
2-3:15 PM: Station Reports
3:15-3:30 Break
3:30-4PM Station Reports (continued)
4:00-4:30PM: Business Meeting.
4:30 PM: End of joint NC1027 + BRD CAP meeting

Thursday September 22, 2011
9:00 to 10:30: Open Forum, in conjunction with AABP BRDC committee and members of BRD CAP. Discussion of questions related to bovine respiratory disease with AABP Convention attendees.

Minutes of business meeting, September 21:

Deb Hamernik, Administrative Advisor to NC-1027/NC-1192, briefly addressed the group. Asked that in the future, extramural funding received be included in annual reports. The project running from 2012-2017 will be identified as NC-1192.

Discussion of location for next years meeting. NC-1027 Technical Committee meeting has been in conjunction with AABP for several years. However, in 2012 AABP will meet in Montreal, Quebec, Canada. International travel is not reimbursed by some universities. Thus the NC-1192 Technical Committee meeting in 2012 will be held in conjunction with Academy of Veterinary Consultants Summer Meeting in Kansas City, MO, Aug 2-4. Exact date, time, and location of Technical Committee meeting TBA.

Discussion of whether AFRI grant recipients who are doing research on BRD will be asked to present at the 2012 meeting. That group joined NC-1027 at 2010 meeting and consensus was that the group meeting was a great success. New collaborations developed between some members of NC-1027 and AFRI awardees through discussions at the meeting. Peter and Margo asked that NC-1027 members email them to confirm their interest in having AFRI awardees join the group at future meetings.

Business meeting ended and Technical Committee meeting adjourned 4:30 PM.

Accomplishments

Accomplishments related to objective 1: To evaluate the prevalence of viral and bacterial agents of respiratory disease by developing, validating and disseminating new state-of-the-art molecular diagnostics for rapid identification of these agents.<br /> <br /> MI evaluated patient-side diagnostic kits for human respiratory syncytial virus (HRSV) detection for use in detecting bovine respiratory syncytial virus (BRSV), using BRSV isolates obtained from GA and other sites. Kits were shown to identify BRSV, with the Tru RSV assay showing the best agreement in preliminary testing. On field samples, the Tru RSV assay was in agreement with RT-PCR 40 of 52 times (k=0.490). There appears to be a problem with false positive results (assuming the RT-PCR is correct). MI is currently working to understand this phenomenon. <br /> <br /> SD and WI continued to report results of surveillance for BRD pathogens in diagnostic laboratory submissions. SD found that principal respiratory pathogens were similar to preceding years with Pasteurella multicida, Mannheimia haemolytica, and cytopathic bovine viral virus diarrhea (BVDV) most frequently isolated. In contrast, WI found BRSV, bovine respiratory coronavirus, P. multocida, and mycoplasmas to be the most common isolates. <br /> <br /> KS developed and refined accurate molecular diagnostic assays for agents involved in BRD. <br /> <br /> OK evaluated bovine coronavirus (BCV) infections in transported commingled calves from auction markets. BCV was found in nasal swabs and lung lavage samples upon entry to a feedlot, and the calves cleared the infections within days of entry. Calves from ranches in a retained ownership program were tested for BCV antibodies upon entry to the feedlot, and those with low virus neutralizing antibodies were more likely to be treated for BRD than those calves with higher BCV antibody levels. There was a wide range of BCV antibody titers among the calves from the respective ranches, indicating variable rates of exposure in the breeding herds. <br /> <br /> OK evaluated bovine herpesvirus-1 (BHV-1) strains including MLV vaccine, reference strains (BHV1.1 and BHV1.2), and isolates from respiratory cases and aborted fetuses, using molecular studies including sequencing of multiple regions of the viral genome. In one group of calves entering a feedlot, the BHV1.2 (genital strain) was identified, and a virus strain from calves collected postvaccination had genomic regions identical to the MLV vaccine strain given at entry. <br /> <br /> Accomplishments related to objective 2: To investigate the basic biology, molecular pathogenesis, and immunopathogenesis of polymicrobial infections including important viral and bacterial agents.<br /> <br /> CA has evaluated mechanisms of enhanced disease related to coinfection with bovine respiratory syncytial virus (BRSV) and Histophilus somni (H.somni). Responses of bronchoalveolar type 2 cells provided by IA to coinfection by BRSV and H. somni were evaluated by identification of genes expression via a bovine microarray. Several genes that were synergistically up-regulated are under further study. Cell viability was also examined after treatments described above. H.somni and BRSV appear to act in synergy at the molecular, cellular, and whole animal levels to cause or enhance respiratory disease. CA also showed that BRSV infection enhanced production of IgE antibodies against inhaled ovalbumin (a prototype allergen) during the acute phase of viral infection; this indicates that BRSV infection may enhance lung inflammation due to environmental allergens. <br /> <br /> NE has continued studies of mechanisms of bovine herpesvirus-1 (BHV-1) latency; research indicated that viral ORF-2 sequences specifically interact with two cellular proteins (Notch 1 and Notch 3) that regulate cell survival and differentiation, and the domains in ORF-2 necessary for inhibiting apoptosis and interfering with Notch1 dependent activation of viral promoters have been identified<br /> <br /> MS continued work on experimental annotation of BRD pathogens by proteogenomic mapping and high throughput RNA sequencing, focusing on Histophilus somni (H. somni). A total of 278 polycistronic operons were identified containing 730 genes. Intergenic regions were also scanned to identify novel expressed genes or small RNAs. More than 150 novel expressed regions with length >50 bp were identified. The RNA-Seq based transcriptome map identified 94 sRNAs in the H. somni genome of which 82 sRNAs were never predicted or reported in earlier studies. We also identified 38 novel potential protein coding open reading frames that were absent in current genome annotation. When compared with the genome sequence of a non-virulent strain 129Pt, a disproportionate number of sRNAs (~30%) were found to be specific to strain 2336 (~18% of the total genome). This observation suggests that a number of the newly identified sRNAs in strain 2336 may be involved in strain-specific adaptations that could include virulence. <br /> <br /> MS and SD evaluated the impact of BVDV on antigen cell presentation. MS used comparative protein profiling approach to elucidate the mechanisms involved in BVDV interference of monocyte and dendritic cell antigen presentation, which allows the virus to avoid effective recognition and elimination by innate and adaptive immune responses. This research confirms that low doses of BVDV significantly affect early apoptotic and oxidative stress mechanisms in cp and ncp BVDV-infected monocytes compared to control cells. SDs studies of the effect of BVDV on dendritic cell antigen presentation are ongoing in collaboration with TX. The culture system for producing mature dendritic cells in culture has been optimized, and studies to evaluate antigen presentation are underway. Preliminary studies are being done with MDBK cells and BVDV strains TGAN and TGAC to optimize the assays to determine the differences in autophagosome trafficking between a cytopathic and noncytopathic virus pair and the best trafficking markers to use for dendritic cells.<br /> <br /> SD undertook studies of innate immunity in the BVDV infected fetus. The livers from fetuses infected at various points in gestation were evaluated, in order to measure the specific immunological effects that occur in the innate immune response of the fetus following BVDV infection and to investigate the role of liver in tolerance in persistent infection. Kupffer cells (liver macrophages) from fetal livers of BVDV PI and control fetuses at 96 days of gestation (21 days post BVDV infection) were isolated and two macrophage immune functions, phagocytosis and antigen presentation, were compared to uninfected controls. Additionally, IHC analysis of fixed tissue was done to assess antigen distribution within the PI liver. There was an increase in the phagocytic activity of the Kupffer cells in BVDV infected fetuses as compared to control fetuses in freshly isolated cells. MHC I expression was increased in persistently infected cultures as well. BVDV antigen was widely dispersed throughout the liver and was first seen in the liver at 82 days but was highest at day 120. <br /> <br /> WI has continued work on the intracellular events in bovine leukocytes exposed to M. haemolytica leukotoxin (LKT). During the above research it was observed that bovine neutrophils release DNA in a time and concentration dependent manner when incubated with M. haemolytica cells or the LKT produced by M. haemolytica. This response is similar to the neutrophil extracellular trap (NET) formation described previously for human neutrophils. NET formation in response to M. haemolytica cells is dependent on LKT; cells that produce native LKT or the lktC mutant LKT elicit NET formation, while cells that produce a truncated inactive LKT do not. NET formation is not elicited by incubation with M. haemolytica LPS. As reported previously for human PMNs, bovine NETs trap and kill a portion of the M. haemolytica cells. NET formation is dependent on PMNs undergoing an oxidative burst. WI has since shown that bovine monocyte-derived macrophages and alveolar macrophages also produce extracellular traps (termed METs) upon exposure to M. haemolytica or its LKT in vitro. As is seen with NETs, M. haemolytica cells are ensnared and a portion of those cells killed within METs. Scientists at WI believe this is the first demonstration of extracellular trap formation by mononuclear phagocytes by any mammalian species. Extracellular traps (NETs and METs) can be remodeled and degraded by DNAse I in bovine serum, and by DNAse II produced by bovine macrophages. These findings suggest that extracellular trap formation is a dynamic process. In preliminary experiments, WI also found that Histophilus somni cells will elicit trap formation by bovine leukocytes in vitro. Experiments are underway to ascertain what component(s) of the H. somnii cells is responsible for this response.<br /> <br /> WI also evaluated the synergistic effects of BHV-1 and M. haemolytica infection in bovine bronchial epithelial cells through gene microarray analysis. 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. Three hundred and fifty five genes exhibited comparable expression profiles following virus or bacterial exposure, suggesting that gene regulation by virus exposure is restricted to a small set of specific targets. Co-exposure to BHV-1 and M. haemolytica altered the viral response in a synergistic or antagonistic manner, consistent with our previous findings. BHV-1 induction of expression of IFN-y, IL-1B, IL-1±, IL-8, and Tnf-± was confirmed by RT-PCR. By comparison, M. haemolytica treatment produced significantly greater inductions (>10 fold) of several genes, including CXCL2, PTX3, IL6, IL1A, SERPINB2, and IL8, compared to BHV-1 alone. Surprisingly, co-exposure to BHV-1 and M. haemolytica resulted in a switch from repression to activation for BHV-1 repressed genes such as Cxcl10. 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. <br /> <br /> Accomplishments related to objective 3: To develop management and prevention strategies that incorporate new vaccines and treatment protocols to combat bovine respiratory disease and reduce economic loss<br /> <br /> GA, FL, KS, IA, and NE collaborated to survey 2500 cow-calf producers in 6 states to determine risk factors for BRD in nursing beef calves. Analysis of the results of the survey is ongoing.<br /> <br /> KS evaluated clinical, behavioral, and pathophysiological data of potential value in detecting the onset and progression of BRD in cattle challenged with Mannheimia haemolytica or Mycoplasma bovis. Studies are ongoing to determine those parameters of diagnostic and prognostic value.<br /> <br /> IA evaluated the genes in Mycoplasma bovis which are related to cytotoxicity in work to characterize M. bovis mutants which can be used in modified-live vaccines to protect cattle from disease due to this pathogen. The cytotoxicity of Mycoplasma bovis for bovine lung epithelial cells was studied using 3 separate methods and a transposon mutant library evaluated to identify M. bovis genes involved in cytotoxicity. Overall, a crystal violet stain method using a bovine lung epithelial cell line proved to be most reproducible, compared to a host cell thymidine uptake or a lactic dehydrogenase release assay. Cytotoxicity was slightly enhanced by addition of glycerol, markedly enhanced when fetal bovine serum was added, and required a high multiplicity of mycoplasmas per cell to be produced. A mutant of M. bovis made defective in genes of a carbohydrate uptake transporter had significantly reduced cytotoxicity than its wild counterpart; this mutant will be the focus of future studies to develop candidate strains for inclusion in modified-live vaccines. <br /> <br /> KS evaluated two different preventive health programs to determine their influence on BRD in the feeding period. Each program consisted of a variety of vaccinations and parasiticide administration. Program 1 had a greater percentage of calves developing BRD compared to program 2 (59.7% vs. 47.8%). Compared to program 2 calves, program 1 calves took more steps each day during the first 28 days of the study, but also spent .more time lying down on certain days during the last 14 days of the study. There were no differences between programs in mortality, case fatality, 1st treatment success or chronicity risks. However, the average daily gain for the program 2 calves (1.23 kg) was greater than program 1 calves (1.16 kg). <br /> <br /> MI continued to work on a bovine viral diarrhea virus (BVDV) eradication program in the Upper Peninsula (UP) of Michigan. Research suggests a minority of herds have PI animals present in them. Elimination of these animals from this low percentage of herds has potential to greatly impact BVDV control in the US. To date (July 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. Our benchmark goal is to have tested for and controlled BVDV in 80% of the beef and dairy herds and 95% of the cattle in the UP by 2012. <br /> <br /> MI evaluated the impact of preweaning BVDV vaccination on health in calves exposed to BVDV PI cattle in the feeding period. Two weeks prior to weaning, all calves were blocked by date of birth and assigned to one of two vaccination groups 1) vaccinated with Bovi-Shield GOLD® FP® 5 or 2) NOT vaccinated with Bovi-Shield GOLD® FP® 5 . At weaning, calves were moved, comingled and housed in one pen. Upon arrival, 4 BVDV PI calves were immediately introduced into the group. Calves were housed with PIs and fed for 220 days. Over the observation period, 22 (33%) steers in the vaccinated group were treated for fever of unknown origin (FUO) at least once compared to 30 (42%) in the unvaccinated control group. There was no affect on response to treatment as measured by the number of times calves needed to be retreated after being pulled for FUO. However, there was a trend for unvaccinated calves to be repulled for new morbidity events. Mortality was low with only 4 (0.5%) steers dying; three from the pre-vaccinated group and one from the unvaccinated control group. Although there was a trend of decreased overall morbidity, retreatment and repull rates, there was no statistical difference between the vaccinated and unvaccinated groups (p=0.25). There was no difference between the pre-vaccinated and non-vaccinated groups in terms of weight or ADG over the course of the observation period. <br /> <br /> OK continued research on immunologically important M. haemolytica outer membrane lipoproteins, in order to characterize the response to vaccination using these immunogens. The specificity and functionality of anti-PlpF antibody responses were demonstrated by marked reduction of complement-mediated killing by blocking killing with rPlpF. PlpF may have vaccination potential against M. haemolytica in cattle. A rapid microtiter plate method for determining serum complement mediated killing of M. haemolytica was developed and evaluated. The immunogenicity of recombinant outer membrane proteins SSA-1, OmpA, OmpP2, and OmpD15 was demonstrated as vaccination of mice and cattle with rOmpA and rSSA-1 stimulated high antibody responses, whereas OmpD15 and rOmpP2 were less immunogenic. <br /> <br /> OK evaluated predictors of feedlot performance in calves from a retained ownership program using serums collected at entry from postweaned calves and tested for antibodies to multiple BRD vaccine immunogens. Calves with increased immunity to BHV-1, BVDV, PI3V, BRSV, M. haemolytica and P. multocida performed better in the feedlot, with less effects clinically to these BRD pathogens and provided greater economic return to the owner. <br /> SD along with TX, CO, and scientists in Illinois, Missouri, and New York, was involved with the Genetics of Feedlot Health Project. This study looked at the relationship between behavior, genetics, nutrition, and microbiologic and immunologic factors and the occurrence of BRD, and the impact on carcass quality. There was a correlation with lower disease in animals with high IgG2. The vaccine study demonstrated a 40% decrease in death loss following challenge with a virulent BVDV 7-1/2 months after vaccination in calves vaccinated at 2-3 days of age in the face of maternal antibody.<br />

Publications

Ammari, M. G., McCarthy, F. M., Nanduri, B., Pinchuk, L.M. 2010. Analysis of Bovine Viral Diarrhea Viruses-infected monocytes: identification of cytopathic and non-cytopathic biotype differences. BMC Bioinformatics. 11 (Suppl 6):S9:1-13.<br /> <br /> Ammari, M., McCarthy, F., Nanduri, B., Pinchuk, G., Pinchuk, L. 2010. Analysis of Bovine Viral Diarrhea Viruses-infected monocytes: identification of cytopathic and non-cytopathic differences. December 2010, CRWAD, Chicago <br /> <br /> D. Amrine, B.J. White, D.E. Anderson, R.L. Larson, D. Mosier, D. Renter. Behavior and clinical changes in calves with induced Mycoplasma bovis pneumonia. American Association of Bovine Practitioners Annual Conference, St. Louis, MO. September 2011. <br /> <br /> Afyalew S, Shrestha B, Montelongo M, Confer AW. Identification and immunogenicity of Mannheimia haemolytica S1 outer membrane lipoprotein PlpF. USDA NIFA Project Directors Meeting, Washington DC, April 2011.<br /> <br /> Aulik, N.A., K. M. Hellenbrand, H. Klos, and C. J. CZUPRYNSKI. 2010. Mannheimia haemolytica and its leukotoxin causes neutrophil extracellular trap (NET) formation by bovine neutrophils. Infect. Immun. 78:4454-446.<br /> <br /> 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.<br /> <br /> <br /> Ayalew S, Shrestha B, Montelongo M, Confer AW. Identification and Immunogenicity of Mannheimia haemolytica S1 Outer Membrane Lipoprotein PlpF. Proceedings, International Pasteurellaceae Conference, Helsinger, Denmark, 2011; No. 41387.<br /> <br /> Ayalew S, Shrestha B, Payton ME, Confer AW. A Rapid Microtiter Plate Method for Determining Serum Complement-Mediated Killing of Mannheimia haemolytica . Proceedings, International Pasteurellaceae Conference, Helsinger, Denmark, 2011; No. 41388.<br /> <br /> K. J. Austin, J. L. Seabrook, T. E. Engle, R. K. Peel, C. M. McAllister, B. W. Brigham, R. M. Enns, R. L. Weaber, H. Van Campen, G. H. Loneragan, J. L. Salak-Johnson, and C.C. L. Chase. The effect of morbidity on feedlot performance and carcass quality in feedlot steers. 2010. J. Anim. Sci. Vol. 88 E-Suppl. 2. P. 10.<br /> <br /> A.H. Babcock, D. G. Renter, B. J. White, S. Dubnicka, H.M. Scott. Temporal distributions of respiratory disease events within cohorts of feedlot cattle and associations with cattle health and performance indices. Prev Vet Med. 2010, 97:198-219.<br /> <br /> L. Berghaus, D. Hurley, R. Berghaus, M. Pence, R. Ellis, J. Saliki, K. Hurley, K. Galland, B.Burdett, S. Nordstrom, A. Woolums. Impact of route and timing of multivalent respiratory vaccination in the face of maternal antibody (IFOMA) on immune responses to booster vaccination at weaning in beef calves. Proceedings of the Conference for Research Workers in Animal Disease (CRWAD), Chicago IL, December 2010<br /> <br /> B. W. Brigham, C. M. McAllister, R. K. Peel, H. Van Campen, R. L. Weaber, G. H. Loneragan, J. L. Salak- Johnson, C. C. L. Chase, E. J. Pollak, and R. M. Enns. The relationship of bovine respiratory disease and carcass ultrasound measures. 2010. J. Anim. Sci. Vol. 88 E-Suppl. 2. P. 41<br /> <br /> N. Cernicchiaro, D.G. Renter, B.J. White, A.H. Babcock, J.T. Fox. Association between weather conditions and bovine respiratory disease in feedlot calves. American Association of Bovine Practitioners Annual Conference, St. Louis, MO. September 2011. <br /> <br /> N. Cernicchiaro, B.J. White, D.G. Renter, A.H. Babcock, L. Kelly, R. Slattery. Effects of transport distance and weight loss during transit on feedlot health and performance. American Association of Bovine Practitioners Annual Conference, St. Louis, MO. September 2011. <br /> <br /> Corbett EM, Grooms DL, Bolin SR. Serological Evaluation of Sentinel Calves In a BVDV Eradication Program. J Vet Diagn Invest. 2011;23(3):511-515<br /> <br /> Corbett EM, Grooms DL, Bolin SR. Evaluation of Skin Samples by RT-PCR Following Immunization with a Modified-Live Bovine Viral Diarrhea Virus Vaccine. Vet Diagn Invest. Accepted November 2010<br /> <br /> Corbett E, Grooms D, Bolin S. Evaluation of skin samples by RT-PCR following immunization with a modified-live Bovine Viral Diarrhea Virus vaccine. Proceedings of the 2010 Conference of Research Workers in Animal Diseases, Chicago, IL, December 5-7, 2010, Abstract # 157.<br /> <br /> da Silva L, Gadreault N, Jones C. 2011. Cytoplasmic localized infected cell protein 0(pICP00 encoded by bovine herpesvirus 1 inhibits beta interferon promoter activity and reduces IRF3 protein levels. Virus Res: In press.<br /> <br /> da Silva L, Jones C. 2011. Infection of cultured bovine cells with bovine herpesvirus1 (BHV-1) or Sendai virus induces different beta interferon subtypes. Virus Res 157:54-60.<br /> <br /> Fulton RW, Step DL, Wahrmund J, Burge LJ, Payton ME, Cook BJ, Burken D, Confer AW. Bovine coronavirus infections in transported commingled beef cattle and sole source ranch calves. Can J Vet Res 75: 191-199, 2010.<br /> <br /> Fulton, R.W., Step, D.L., Wahrmund, J., Burge, L.J., Eberle, R., dOffay J.M., Confer A.W., Cook, B.J., Burken, D., Richards, C., Payton, M.E.: Bovine Coronavirus Infections in Transported Commingled Beef Cattle and Sole Source Ranch Calves. American Association of Bovine Practitioners Proceedings. August 19-21, 2010. Albuquerque, NM..<br /> <br /> Fulton, R.W., dOffay J.M., Burge L.J., Eberle R., McCormack, K., Step D.L., Wahrmund J., Holland B.: Bovine Herpesvirus-1: Molecular Studies of Isolates from Clinical Cases Characterizing Field Isolates and Vaccine Strains. 53rd Annual Meeting of AAVLD, November 11-17, 2010. Minneapolis, MN.<br /> <br /> Fulton, R.W., Confer, A.W., Joint Presentation. Gold Standards for Bovine Respiratory Diagnosis, Do They Exist? Laboratory Tests, the Good, the Bad, the Ugly. Academy of Veterinary Consultants Meeting, March 31-April 2, 2011, Oklahoma City, OK.<br /> <br /> <br /> Gaudreault N and Jones C. 2011. Regulation of promyelocytic leukemia (PML) protein levels and cell morphology by bovine herpesvirus 1 infected cell protein 0 (bICP0) and mutant bICP0 proteins that do not localize to the nucleus. Virus Res 156:17-24.<br /> <br /> Geertsema RS, Zekarias B, La Franco Scheuch L, Worby C, Russo R, Gershwin LJ, Herdman DS, Lo K, Corbeil LB. IbpA DR2 subunit immunization protects calves against Histophilus somni pneumonia. Vaccine. 2011 Jun 24;29(29-30):4805-12. Epub 2011 May 8. <br /> <br /> Gershwin LJ, Anderson ML, Wang C, Berghaus LJ, Kenny TP, Gunther RA. Assessment of IgE response and cytokine gene expression in pulmonary efferent lymph collected after ovalbumin inhalation during experimental infection of calves with bovine respiratory syncytial virus. Am J Vet Res. 2011 Jan;72(1):134-45. <br /> G. Hanzlicek, B.J. White, D. Renter , D. Anderson, R. Larson. Association between the prevalence of Mollicutes and Mycoplasma bovis and health and performance in stocker calves. Vet. Rec. 2011, 168(1):21. Epub 2010 Dec. 17.<br /> <br /> G.A. Hanzlicek, B.J. White, D.G. Renter, D. Blasi. A field study evaluating health, performance, and behavior differences in crossbred beef calves administered different vaccine-parasiticide product combinations. Vaccine. 2010, Aug. 28(37):5998-6005. Epub 2010 July 14. <br /> <br /> G. Hanzlicek, D. Renter, B.J. White, B.A. Wagner, D.A. Dargatz, M.W. Sanderson, H.M. Scott, R. Larson. Management practices associated with the rate of pre-weaning calf respiratory disease: results from a national survey of U.S. cow-calf operations. American Association of Bovine Practitioners Annual Conference, St. Louis, MO. September 2011. <br /> <br /> Kisiela DI, Aulik NA, Atapattu DN, CZUPRYNSKI, CJ. 2010. N-terminal region of Mannheimia haemolytica leukotoxin serves as a mitochondrial targeting signal in mammalian cells. Cell. Microbiol. 12:976-987.<br /> <br /> Krunkosky TM, B. Heins, L. Berghaus, M. Ard, C. Jarrett, A. Woolums.. Morphology of Primary Bovine Bronchial Cultures from Live Calves and Cellular Responses to Bovine Respiratory Syncytial Virus. Am J Resp Crit Care Med. 181:B182, 2010.<br /> <br /> Kumar, R., B. Nanduri, J. Watt, A. Cooksey, and M. L. Lawrence. 2010. High throughput transcript mapping of Histophilus somni 2336 by deep sequencing. Conference for Research Workers in Animal Disease, Chicago, Illinois.<br /> <br /> Lee, N., Rosenbusch, R. F. Characterization of an ATP-binding cassette transport system responsible for nucleoside uptake in Mycoplasma bovis. Poster G-1271, 110th General Meeting of the American Society for Microbiology, San Diego, CA, 2010.<br /> <br /> Leyh, R.D., Fulton, R.W., Stegner, J.E., Goodyear, M., Witte, S., Taylor, L.P., Johnson, B.J., Step, D.L., Ridpath, J.F., Holland, B.P.: Fetal Protection in Heifers Vaccinated with a Modified Live Virus Vaccine Containing Bovine Viral Diarrhea Virus Subtypes BVDV1a and BVDV2a and Exposed During Gestation to Cattle Persistently Infected with BVDV Subtype 1b. American Journal for Veterinary Research 72: 367-375, 2011. <br /> <br /> Morarie, S. Mediger J, Braun L, Chase C, Smirnova N, Hansen T. Fetal immunological effects and liver tolerance following persistent bovine viral diarrhea virus infection. Abstract 126P. 91st annual meeting of Conference of Research Workers in Animal Disease, Chicago, IL, December 5-7, 2010. <br /> <br /> <br /> Neill J, Schefers J, Chase C, Ridpath J. Vaccination of cattle persistently infected with BVDV does not cause a change in the consensus sequence of the structural proteins of the viral quasispecies. 53rd Annual Joint Meeting American Association of Veterinary Laboratory Diagnosticians (AAVLD)/United States Animal Health Association (USAHA). November 11-17, 2010, Minneapolis, MN<br /> <br /> Panciera RJ, Confer AW. Pathogenesis and pathology of bovine pneumonia. Vet Clin North Amer Food Anim Prac 26: 191-214, 2010<br /> <br /> Pinchuk, L., Ammari, M., McCarthy, F., Nanduri, B. 2010. Proteomic analysis of Bovine Viral Diarrhea Virus-infected monocytes. December 2010, CRWAD<br /> <br /> Rose-Dye TK, Burciaga-Robles LO, Krehbiel CR, Step DL, Fulton RW, Confer AW, Richards CJ. 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Impact Statements

1. The development of rapid and accurate diagnostic tests for BRD agents will help veterinarians and producers confirm the cause of BRD outbreaks more quickly, allowing more rapid and targeted treatment and management efforts to better ameliorate the impact of BRD in affected cattle.
2. Surveillance for new, emerging, and re-emerging pathogens in field cases and outbreaks of BRD will help stakeholders understand which pathogens should be the focus of control and prevention efforts for different cattle operations.
3. The comparison of the outcome of different preventive health programs on calf health in the feeding period will help veterinarians and producers identify the optimal approach to managing cattle to prevent BRD.
4. Identification of responses of alveolar epithelial cells to BRSV and H. somni coinfection via microarrays will provide new cellular targets for therapeutic or preventive strategies to decrease disease associated with viral-bacterial coinfection in BRD.
5. Results of the studies of BHV-1 latency will provide new molecular targets that may be amenable to manipulation to counteract herpesvirus latency or reactivation and thus prevent disease due to this virus.
6. The results of the survey of risk factors for nursing beef calf pneumonia will help veterinarians and producers to develop management strategies to decrease calf illness and improve calf productivity in cow-calf herds where calf respiratory disease is a problem.
7. Virulence of M. bovis strains and single-hit transposon mutants has been studied to lay the foundation for rational design of gene-deleted mutants that could be used as live avirulent vaccine strains to protect cattle against respiratory disease involving M. bovis.
8. The BVDV eradication program being carried out in the UP of Michigan will provide proof-of-concept data and a valuable model for other regions of the U.S. that wish to limit the impact of this serious pathogen through regional eradication.
9. Tests of methods to control the impact of BVDV PI cattle on the health of in-contact cattle will provide information to veterinarians and producers so they are able to determine which control measures are most useful to preserve health and prevent disease in cattle at risk for exposure to BVDV PI cattle.
10. Proteogenomic mapping and high throughput RNA sequencing of H. somni will improve the knowledge of the mechanisms by which this pathogen causes disease, and will provide targets for new drugs and vaccines to prevent disease due to this agent.
11. The evaluation of the impact of BVDV on antigen presenting cell function will provide foundational knowledge regarding the means by which BVDV impairs host immunity to infection with BVDV and also other pathogens which often participate with BVDV in dual infections of cattle. This knowledge will form the basis for development of methods to counteract the well-recognized immunosuppression caused by BVDV.
12. The studies of the immunogenicity of M. haemolytica outer membrane proteins will provide information needed to develop newer generation vaccines for use to protect cattle against disease due to this common respiratory pathogen.
13. The development of a rapid microtiter plate method for determining serum complement mediated killing of M. haemolytica will provide researchers with a useful tool for confirming whether new vaccines and immune stimulants are effective at inducing protective immunity in cattle.
14. The evaluation of the relationship between serum antibody titers to respiratory pathogens at arrival and health outcomes in feedlot cattle will provide veterinarians and producers with guidelines for management of cattle in ways that can decrease BRD during the feeding period.
15. The studies of the response of immune cells in the fetal liver to prenatal BVDV infection will provide foundational knowledge regarding the means by which BVDV establishes an immunotolerant state in the infected fetus, which is necessary for the generation of persistently infected (PI) animals which are a major source of infection for other cattle. This work will also expand basic knowledge regarding how the bovine immune system develops and functions, which will improve the ability of scientists to produce effective vaccines and immunostimulants.
16. The studies of NETs and METs produced by immune cells in response to BRD pathogens have revealed new mechanisms by which the bovine immune response fights pathogens. This improves the knowledge of scientists and veterinarians regarding immune system function, and provides a baseline for future research to maximize immunity to respiratory and other infections in cattle.

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