Brief Summary of Minutes of Annual Meeting

The meeting was called to order at 8:10 A.M. by the Chair, Dr. Connie Gebhart. Linda Saif, with a second by Don Robertson, moved for approval of the 2004 minutes. The minutes were approved by a unanimous vote. Connie Gebhart announced that dues were being accepted by her for all NC1007 members in attendance. Considerable discussion on the amount of dues followed and is summarized below: Lynn Joens suggested that dues should be higher to cover the costs of the Gastroenterology awards presented at the CRWAD since the awards are already less that those given by other award committees. Dave Frances indicated that $50.00 would cover the cost of the meeting room and refreshments. More discussion followed regarding the lack of attendance by some stations and whether all stations should pay dues if a representative does not attend the meeting. Richard Isaacson moved that NC1007 should obtain industrial support for the Gastroenterology awards at the CRWAD meeting. In the discussion that followed, it was suggested that we ask for $thousands, and not $hundreds, to support these awards and meetings such as the Rushmore Conference. Richard Isaacson and Lynn Joens volunteered to draft a white paper that will explain and justify industrial support of awards and meetings sponsored by NC1007 such as the Rushmore Conference. The proposal was adopted with unanimous support. Lynn Joens then moved to thank Dave Francis for all his hard work and the success of the Rushmore Conference, which was met with enthusiastic approval. The NC1007 administrative advisor, Bert Stromberg made several comments. Bert also complemented Dave Francis and all those at South Dakota State University who contributed to the success of the Rushmore conference. He reported that the project reporting format had changed and SAE form 422 must be filed by him within 90 days after the current meeting. The report should include: 1) accomplishments, and 2) impacts of the work done by members of NC1007. Station reports should be included as an appendix. Publications should be separated into: 1) those from each participating station and 2) publications resulting from collaborative efforts. Dr. Stromberg expressed concern about the low number of publications resulting from collaborative stations in NC1007. He also informed us that NC1007 will sunset in 2007, if not renewed. A brief letter is needed by the end of 2005 or by the end of February at the latest, 2006, which should include a vision of 3-6 sentences and at least 3 objectives for the future. This information will be distributed to all Experiment Station Directors in an attempt to increase the number of members and station participation in NC1007. The final renewal proposal must be completed by November of 2006. Richard Isaacson expressed concern that it is difficult to show the impact of the work done by NC1007, which is also supported by Hatch projects that pay salaries of principal investigators. More important, the progress report and impact statements do not indicate the amount of infrastructure that is necessary to do the work. Since the issue of awards was not resolved before Bert Strombergs comments, discussion continued on the matter. Lynn Joens raised the issue whether industry should be asked to support travel , banquets and other functions associated with the mission of NC1007. The discussion returned to the nature of the awards presented at the CRWAD meeting. The following issues were resolved: 1. The award will retain the NC1007 name. 2. Industry partners will be recognized in the program at presentation ceremonies. 3. The amount given each year will fluctuate depending on available funds. 4. The awards for graduate students will be included and highlighted in the NC1007 renewal proposal. The issue not resolved was whether NC1007 should present a Food Safety award at the CRWAD meeting. Since Richard Isaacson is the Food Safety Award committee and Lynn Joens is in charge of the Gastroenterology awards, each will talk with their respective committees and come back with a recommendation at the 2006 meeting on whether NC 1007 should present a Food Safety Award. Dave Francis moved that Connie Gebhart appoint a committee to write a brief synopsis of the Rushmore conference so that it can be included in the SAE 422 report and in support of a future meeting to be proposed in the renewal. The committee members are Lynn Joens (Chair), Dave Francis, and Richard Isaacson, with input by Roy Curtiss. Officers for the coming two years were elected by acclamation. Tom Besser will be Chair for the next two years and Don Robertson will serve as Secretary. Discussion continued on the focus of the NC1007 renewal proposal. Linda Saif suggested that the focus should be on Common Pathogenic Mechanisms of Zoonotic and Emerging Diseases. This focus could take advantage of much of the expertise in NC1007 on viruses, Camplobacter, Salmonella and diarrheagenic E. coli. Dave Francis suggested that we should not forget the need for better animal models in the renewal proposal. Other suggested topics include the importance of the turnover rate of intestinal epithelial cells and the impact on host-parasite relationships. Discussion continued to return to innate resistance and its impact on changes in virulence due to interspecies transfer coupled with the use of antibiotics in this process. Richard Isaacson suggested that one objective of the NC1007 renewal proposal should be to organize a colloquium that will identify needs and gaps in studies on Emerging Diseases and Effects of Interspecies Transfer on Virulence based on his positive experience with the Food Safety meeting recently held in Scotland. Summary of action items from meeting. 1. A summary of the Rushmore conference will be drafted and submitted to a journal such as the Journal of Infectious Disease, which routinely publishes such articles. 2. Richard Isaacson will approach the Academy of Microbiology to determine their level of interest in supporting and organization of a meeting dealing with Common Pathogenic Mechanisms of Zoonotic and Emerging Infectious Diseases including changes of virulence due to interspecies transfer and the importance of innate immunity. 3. Tom Besser will coordinate drafting a vision statement and objectives for the NC1007 renewal and will call a meeting of those committee members present at the 2005 CRWAD meeting in St. Louis. Tom Besser moved to adjourn the meeting with a second by Don Robertson.

Accomplishments Objective 1  Define mechanisms of pathogen-host-environmental interactions in enteric and food borne diseases. Campylobacter species AZ has shown that many C. jejuni isolates recovered from non-poultry animal sources have the capacity to cause disease if transmitted to humans. They further assessed the in vitro and in vivo pathogenicity of C. jejuni isolates from broiler chickens and reported the presence of a novel pilus expressed by C. jejuni under biofilm conditions. Clonal strain types of C. coli carrying multiple antimicrobial resistance characterized by PFGE (two enzymes), plasmid profiles and resistance genotyping have been identified as accounting for most of the multiply drug resistant thermophilic Campylobacter sp on WA cattle farms, and related clonal types have been identified on CA cattle farms. Salmonella species AZ reported that an emerging pathogen, S. Newport, has higher survival rates in oysters than E. coli and S. Typhimurium. WA is continuing S. enterica surveillance with collection of isolates from both bovine and human sources from around WA. Antimicrobial resistance profiles are used as a means to putatively match isolates and this latter subset is further evaluated using PFGE. To date numerous matches between strains originating from human and bovine sources have been identified. Lawsonia species MN has demonstrated expression and immunogenicity of ten proteins encoded by the genome of L. intracellularis. Escherichia coli NE is evaluating disruption of enterotoxin genes of enterotoxigenic Escherichia coli by allelic exchange using lambda Red-mediated recombineering. They have optimized the lambda Red recombineering technology for use in ETEC, as demonstrated by the precise disruption of the heat-stable enterotoxin-b (estB) and heat-labile enterotoxin (eltAB) genes, results which encourage further use of this technology in studies aimed at the elucidation of gene function. WA has continued research on the role of the bovine recto-anal junction (RAJ) as a site of colonization for E. coli O157:H7 and how this host-bacterial interaction is involved in the epidemiology of E. coli O157:H7 within cattle populations. Final data analysis confirmed the RAJ colonization and super-shedder findings that were presented in the previous NC-1007 progress report. These were complemented by further analyses including PFGE characterization and comparison studies of isolates. RAJ and fecal isolates matched by animal and date are usually identical or very closely related, suggesting that the RAJ is a significant source of fecal strains. Final data analysis has been performed by WA for another E. coli O157:H7 RAJ project that aimed to examine if bovine immunogenetic parameters were associated with E. coli O157:H7 colonization susceptibility. No associations between the likelihood of an individual heifer to be either E. coli O157:H7-positive or negative and the measured immunological parameters were noted. A project investigating the prevalence and public health risk of non-O157 STEC in foods has been completed. A total of 1440 cattle and 1079 food samples originating from within WA State were examined. From these, a total of 126 bovine and 110 food STEC isolates were characterized, along with 121 regionally and temporally-matched human clinical isolates. A large proportion of bovine-derived food samples had evidence of STEC contamination, based on presence of stx or isolated STEC, indicating a notable likelihood for human exposure, even with post-harvest processing procedures in place. Helicobacter species NE demonstrated that the cytolethal distending toxin B sub-unit of H. hepaticus is a Ca2+- and Mg2+-dependent neutral nuclease. They also showed that it localizes to the nucleus and is the main determinant for intoxication of eukaryotic cells. Enteric Viruses A porcine enteric calicivirus, Cowden strain, can be propagated in a porcine kidney continuous cell line (LLC-PK) in the presence of bile acids in the cell culture medium. OH generated a full-length cDNA infectious clone of this Cowden strain and the capped RNA transcripts derived from this clone were infectious when transfected into LLC-PK cells. The recovered virus retained its ability to infect piglets when administered by the oral route and showed an attenuated phenotype similar to that of the tissue culture-adapted parental virus. To determine the occurrence of human and animal enteric caliciviruses in U.S. market oysters, OH surveyed regional markets. Oysters were collected from different growing areas in the Atlantic and Pacific Oceans and the Gulf coast during summer 2002 and winter of 2002-2003. Animal enteric caliciviruses were detected in 15 samples (33%) by RT-PCR. Bovine noroviruses were detected in 2 samples. Different seasonal and state distributions were detected for human and animal enteric caliciviruses. The presence of animal enteric caliciviruses was associated with states (Oregon, Washington) with major livestock production. Objective 2  Develop and improve diagnostics, treatment, and preventative measures for enteric and food borne diseases. Campylobacter species NE is characterizing a novel Campylobacter cytolethal distending toxin from C. hyointestinalis subsp. hyointestinalis isolated from humans and pigs. Comparative phylogenetic analysis of partial nucleotide and amino acid sequences of cdtB revealed that human and porcine C. hyointestinalis subsp. hyointestinalis formed a cluster distinct from other ECS. Light and confocal laser scanning microscopic examinations of HeLa and INT-407 cells incubated with whole-cell lysates of C. hyointestinalis showed characteristic cytoplasmic and nuclear enlargements indicative of CDT activity. NE, in collaboration with MN, IA, and SC, found that US porcine C. coli isolates are negative for cytolethal distending toxin activity. They examined the reference C. coli strain ATCC 49941 and 40 C. coli strains isolated from pigs on commercial farms in Iowa (n=10), Minnesota (n=10), Nebraska (n=10), and North Carolina (n=9) for the presence of cdtB gene and CDT activity. The cdtB gene was present in all of the strains by PCR amplification of a partial sequence using degenerate and gene-specific oligonucleotide primers. Co-incubation of either HeLa or H407 cells with whole-cell lysates obtained from each strain did not elicit cellular changes characteristic of CDT activity, as determined by either examination using light microscopy or quantitative G2/M phase cell cycle arrest using flow cytometry. Group A Rotavirus IL is investigating receptors as therapeutic approaches to control rotavirus infections. Results suggest that specific oligosaccharide/isoflavone mixtures or profiles may be engineered to provide a deliverable nutriceutical approach to the treatment and prevention of rotavirus disease across susceptible animal species and humans. Cryptosporidium IL has determined the cryptosporidium oocysts are effectively retarded from overland transport by vegetative filter strips (VFS) and that the mechanism of this retardation is specific adhesion to the clay particles of the soil that occurs as a consequence of reduced flow over a vegetated surface as compared to bare soil. Lawsonia species MN evaluated various methods of biochemical augmentation and modification of culture conditions for enhanced growth of L. intracellularis in cell culture. Enhancement of growth in various types of cell cultures (cell types; monolayer and suspension) was evaluated. We also attempted to establish appropriate culture conditions for axenic growth of Lawsonia intracellularis. Despite these efforts, L. intracellularis continues to be an obligate intracellular organism, culturable only in cells in vitro. MN developed an enzyme-linked immunosorbent assay for the diagnosis of proliferative enteropathy in pigs. Analysis of variable number tandem repeat regions (VNTR) in the L. intracellularis genome by MN has provided a means for the differentiation of isolates. This method was demonstrated to be stable, reproducible, and applicable to samples from field outbreaks of proliferative enteropathy. Escherichia coli NE is investigating the role of the Tir protein in Escherichia coli O157:H7 intestinal colonization of adult cattle. Their results suggest that serologically detectable responses to Tir are associated with the level of intestinal infection; however, more studies will be required to determine the relative importance of Tir for E. coli O157:H7 colonization of the adult bovine gastrointestinal tract. NE is evaluating vaccination against type III secreted proteins as a strategy to control Escherichia coli O157:H7 in cattle. In commercially fed cattle, the two-dose vaccine regimen reduced the probability of E. coli O157:H7 colonization of the terminal rectum, and reduced pen-level contamination. Collectively, their clinical trials provide scientific evidence that vaccination with type III secreted proteins is an effective pre-harvest intervention strategy for the control of E. coli O157:H7 in feedlot cattle. Brachyspira species NE is examining B. pilosicoli for the presence of penicillin-binding proteins (PBPs) in comparison with known PBPs of E. coli. Five PBPs of slightly different molecular masses (94-, 62-68-, 42-50-, 25-, and 22-kDa) were identified in a human and two porcine B. pilosicoli strains. Spirochetes that were identified as B. pilosicoli were found by NE to be present in the ceca of 7.5- to 18-week-old commercial turkeys with cecal spirochetosis and typhlitis. Enteric Viruses OH performed a prevalence study of porcine noroviruses and GIII sapoviruses in US swine using newly developed RT-PCR-hybridization assays. The porcine noroviruses were detected exclusively from finisher pigs in 4 of 7 farms and 1 slaughterhouse with an overall prevalence of 20% in finisher pigs. Porcine GIII sapoviruses were detected from all ages of pigs. The prevalence of porcine GIII sapoviruses was 62% overall, lowest in nursing pigs (21%) and highest in post-weaning pigs (83%). Mixed infections of noroviruses and sapoviruses were common in finisher pigs with an overall prevalence of 27% among norovirus- or sapovirus-positive pigs. Two types of BEC have been identified in cattle: the bovine noroviruses (BoNoV) that belong to Genogroup III NoVs and the NB-like BEC more similar to lagoviruses and sapoviruses and which may represent a new calicivirus genus. OH evaluated antibody responses to BoNoV virus-like particles (VLPs) and protection against challenge with virulent homologous BoNoV in gnotobiotic calves. Virus shedding was detected post-challenge on days 1-5 by antigen-capture ELISA. By RT-PCR, virus was detected from all challenged calves for 1-10 days post-challenge. No association between serum and fecal immune responses and protection was observed except in calves immunized intranasally with VLPs in the presence of mLT that had fecal IgA antibodies to BoNoV and showed partial protection to diarrhea after challenge. Bovine coronavirus (BCoV) is a pneumoenteric, cultivable, enveloped, single-stranded RNA virus of positive polarity in the Coronavirus genus within the Coronaviridae family and the Nidovirales order. OH compared the shedding patterns and antibody responses to BCoV in two groups of feedlot calves: those shipped from a single ranch versus those from a sale barn and to evaluate the role of BCoV in disease and on performance. Results suggest that the replication and shedding of BCoV may start in the upper respiratory tract and spread to the gut with viral transmission via the nasal or fecal/oral routes. Vaccination of calves with an effective BCoV vaccine, pre-shipping to feedlots, to induce ELISA serum IgG antibody GMT of >1860, may provide protection against BCoV infection and its combined effects with other pathogens and factors in the induction of respiratory disease. Objective 3  Provide training and continuing education opportunities and dissemination of information to students, producers, consumers, veterinarians and diagnostic laboratories. Minnesota Principle Investigators, Graduate and Post-Graduate Students, and Veterinary Students involved in the project have given presentations and updates on proliferative enteropathy at various scientific, veterinary, and diagnostic meetings in the previous year; including the Conference of Research Workers in Animal Disease, the Leman Swine Conference, ANAPORC (Spain), the Rushmore Conference (sponsored by NC-1007) and the American Association of Swine Veterinarians. They have disseminated new information, reagents, and procedures to producers, industries, veterinary diagnostic laboratories and veterinarians. In addition, they have presented an international workshop, including theoretical lectures and practical laboratories, on the diagnosis and control of proliferative enteropathy in pigs. Nebraska Information on E. coli O157:H7 research, which included the results of clinical trials with the vaccine and direct-fed microbial was disseminated to the public in the form of meetings with producers, veterinarians, and food service industries. Groups in which extension presentations were given included: private meat processing groups, food service industry representatives, the Canadian Cattlemen's Association, Ontario Cattlemens Association, and the National Cattlemens Beef Association. Information on a systems approach to controlling neonatal calf diarrhea was presented in extension programs to cattle producers and veterinarians in Nebraska, South Dakota and Kansas. A lecture was provided on animal production food safety to an undergraduate Foods and Sanitation course. Washington MicroPad, A European FP5 Program, Molecular ecology workshop entitled, Detection of microbial biodiversity in environmental samples, Camerino, Italy, 19-21 September 2005. Call, Assessing intra-specific genetic diversity for bacteria and genetic diversity between plasmids using DNA microarrays. WA-DOE, Colville River TMDL Advisory Group, Colville, WA, 19 July 2005. Call, Microbial source tracking: an update on genetic markers, presentation. U.S. Environmental Protection Agency, Cincinnati, OH, Feasibility of Using DNA/RNA Microarrays and Related Technologies for High Through-put Detection of Waterborne Pathogens, 22-23March 2005. Call, A Primer on Microarrays and the Prospects and Challenges for Pathogen Detection WA State Beef Commission Board Meeting invited speaker. Leavenworth, Washington. 11th August 2004. The role of recto-anal junction E. coli O157:H7 intestinal colonization in influencing feedlot cattle E. coli O157:H7 dynamics. Washington Cattlemans Association General Meeting. Pasco, Washington. 12th November 2004. The Role of Super-shedders in the Epidemiology of E. coli O157:H7 in Feedlot Cattle. Cattle Industry Meeting. San Antonio, Texas. 4th Feb 2005. Dynamics of recto-anal junction colonization and associated fecal shedding of Escherichia coli O157:H7 in feedlot cattle. Beef Industry Safety Summit. Orlando, Florida. 20th April 2005. Dynamics of recto-anal junction colonization and associated fecal shedding of Escherichia coli O157:H7 in feedlot cattle. Pacific Northwest Food Safety Farm to Table Conference 2005. Campylobacter in the Food Supply. Moscow ID. 05/25/05.

Publications 2005 AZ Thesis & Dissertations Law, B.F. Assessment of the Pathogenicity of Campylobacter jejuni from Broiler Chickens. Doctorial Dissertation, University of Arizona, Tucson, AZ. 2005. Reeser, Ryan. Campylobacter jejuni: Characterization of a microbial Biofilm on Abiotic Surfaces. Masters Thesis, University of Arizona, Tucon, AZ. 2005. Refereed Publications Lee, M.K., S.J. Billington, L.A. Joens. Potential Virulence and Antimicrobial Resistance in Campylobacter jejuni Isolates Obtained from Food and Companion Animals. Foodborne Path. Dis. 1:223-230. 2004. Brands, D.A., A.E. Inman, C.P. Gerba, C.j. Mare, S.J. Billington, L. Saif, J.F. Levine, L.A. Joens. The Prevalence of Salmonella spp. In U.S. oysters. App. Env. Mic. 71:893-897. 2005 Brands, D.A., S.J. Billington, J.F. Levine, L.A. Joens. Genotyping and Resistance Determination of Salmonella Newport Isolates Obtained from U.S. Market Oysters. Foodborne Path. Dis. 2:111-114. 2005. Book Chapters Joens, L.A.: Campylobacter: Molecular and Cellular Biology by M.E. Konkel. Chapter 22, p 415-420. Horizon Press. 2004. IL Kuhlenschmidt MS, Rolsma MD, Kuhlenschmidt T, Gelberg HB. Characterization of a Porcine Enterocyte Receptor for Group A Rotavirus. In: Mechanisms in the Pathogenesis of Enteric Diseases. ed., Paul PS, Francis DH, Benfield DA., pp. 135-143, Plenum Press, New York, N.Y., 1997. MD Rolsma, TB Kuhlenschmidt., HB Gelberg and MS Kuhlenschmidt (1998) Structure and Function of a Porcine Enterocyte Ganglioside Receptor for Group A Rotavirus. Journal of Virology 72:9079-9091. TB Kuhlenschmidt. B Hanafin and MS Kuhlenschmidt (1999) Sialic Acid Dependence and Indpendence of Group A Rotavirus. Adv. In Experimental Biology and Medicine 473:309-317 Gelberg HB, Thulin JD, Kuhlenschmidt MS. Application of Intestinal Xenografts to the Study of Enteropathogenic Infectious Disease. In: Mechanisms in the Pathogenesis of Enteric Diseases. ed., Paul PS, Francis DH, Benfield DA., pp. 31-35, Plenum Press, New York, N.Y., 1997. Rolsma MD, Kuhlenschmidt TK, Gelberg HB, Kuhlenschmidt MS. 1998 Structure and function of a ganglioside receptor for porcine group A rotavirus. J. Virol. 72:9079-9091) Moreno B, Bailey BN, Luo S, Martin MB, Kuhlenschmidt M, Moreno SN, Docampo R, Oldfield E. (2001) (31)P NMR of apicomplexans and the effects of risedronate on Cryptosporidium parvum growth. Biochem Biophys Res Commun. 284:632-637. J Trask, S. McLaughlin, MS Kuhlenschmidt and P. Kalita 2004 Overland and Near-surface Transport of Cryptosporidium parvum. Environ. Qual. 33:984-983. Johnson, JK, Schmidt, Gelberg, HB, and Kuhlenschmidt MS. 2004. Microbial Adhesion of Cryptosporidium parvum sporozoites: Purification of an Inhibitory Lipid from Bovine Mucosa. J. Parasitology 90:980-990. Wetzel DM., Schmidt J, Kuhlenschmidt MS., Dubey JP, and Sibley LD. 2005.Gliding motility leads to active cellular invasion by Cryptosporidium parvum sporozoites. Infection and Immunity 73:5379-5387. Abstracts: Johnson JK, Kuhlenschmidt MS, Gelberg HB. Glycoconjugates and plasma membrane vesicles inhibit Cryptosporidium parvum sporozoite-host cell recognition. Vet Pathol 34:509, 1997. JK Johnson, HB Gelberg, and MS Kuhlenschmidt (1999) - Partial Purification of a Cryptosporidium parvum Sporozoite Host Cell Receptor from Plasma Membrane Vesicles. Second Annual Conference on New and Re-Emerging Infectioius Diseases. JK Johnson, HB Gelberg and MS Kuhlenschmidt (1999) - Partial Purification of a Cell Surface Receptor for Cryptosporidium parvum sporozoite Attachment. Proceedings: American College of Veterinary Pathologists, 50th Annual Meeting, Chicago, IL. MS Kuhlenschmidt, JK Johnson, and HB Gelberg. 1998. Glycoconjugates and Plasma Membrane Vesicles Inhibit Cryptosporidium parvum Sporozoite-Host Cell Recognition. Second International Rushmore Conference on Mechanisms in the Pathogenesis of Enteric Disease. JJ Ward, RD Smith, and MS Kuhlenschmidt (1999) Evaluation of the Premier Cryptosporidium by Meridian Diagnostics for the detection of C. parvum in dairy cattle. Conference for Research Workers in Animal Diseases Proceedings. JJ, Smith RD, and Kuhlenschmidt, MS (1999).Critical control points for reducing environmental contamination with Cryptosporidium parvum oocysts from dairy cattle, Proceedings, 9th Symposium of the International Society for Veterinary Epidemiology and Economics, Aug 6-11, 2000; Breckenridge, CO. JJ Ward, Smith RD, and Kuhlenschmidt, MS. (2000) Simulation model to predict environmental contamination with Cryptosporidium parvum oocysts from a dairy herd,Proceedings, 81st Annual Meeting of the Conference of Research Workers on Animal Diseases (CRWAD). Nov 12-14, 2000; Chicago, IL. Trask, J. R., P. K. Kalita, M. S. Kulhenschmidt, R. D. Smith, and T. L. Funk. 2001. Overland and Near-surface Transport of Cryptosporidium parvum. ASAE Annual International Meeting 01-2104: 1-14. JJ Ward, RD. Smith, MS Kuhlenschmidt. 2002. Evaluation of Factors Contributing to Environmental Contamination with Cryptosporidium parvum from a Dairy Herd. International Conference on Emerging Infectious Diseases, March 2002, Atlanta, Ga. Salto, M.L., Kuhlenschmidt, T., Kuhlenschmidt, M., de Lederkremer, R. M., and Docampo, R. 2004. Lipid composition of acidocalcisomes of Trypanosoma cruzi. 7th Annual Conference on New and Re-Emerging Infectious Diseases. Urbana, April 15-16, p. 23. http://www.cvm.uiuc.edu/idc/ Ochonicky K, Donovan, S, Kuhlenschmidt T. and Kuhlenschmidt M. 2005. Inhibitory effects of human and porcine milk oligosaccharides on sialic acid dependent and sialic acid independent strains of rotavirus. American Dairy Science Association Annual Meeting, Cincinatti, OH, July 24-28, 2005. Ochonicky K, Donovan S. Kuhlenschmidt T, Jimenez-Flores R, Kuhlenschmidt M. 2005. Inhibitory activity of bovine milk fat globule membrane against sialic acid-dependent and independent strains of rotavirus American Dairy Science Association Annual Meeting, Cincinatti, OH, July 24-28, 2005. Andres A., Donovan S.M., Kuhlenschmidt T.B., Kuhlenschmidt M.S. 2005. Isoflavones at concentrations present in soy-based infant formula inhibit rotavirus infectivity in vitro. The FASEB Journal 2005;19: A446. MN Book Chapter S. McOrist and Gebhart, C.J. 2005. Proliferative Enteropathies. In B. Straw, et al, (ed.), Diseases of Swine, Ninth Edition. Blackwell Publishing, Ames, IA. Journal Article Wattanaphansak, S., C.J. Gebhart, M. Olin, and J. Deen. 2005. Measurement of the viability of Lawsonia intracellularis. Can. J. Vet. Res. (in press). Abstracts Beckler, D.C., V. Kapur and C.J. Gebhart. 2004. Molecular epidemiologic typing of Lawsonia intracellularis. Proc. 85th Conf. Res. Workers An. Dis., Chicago, IL, November 14-16. Nuntaprasert, A, K. Kaur, C.J. Gebhart and V. Kapur. 2004. Expression and purification of a flagellar protein of Lawsonia intracellularis. Proc. 85th Conf. Res. Workers An. Dis., Chicago, IL, November 14-16. Wattanaphansak, S. , K. Kinsley, J. Deen and C. Gebhart. 2004. Comparative agreement of indirect fluorescence antibody test and immunoperoxidase monolayer assay for serological diagnosis of field cases of porcine proliferative enteropathy. Proc. 85th Conf. Res. Workers An. Dis., Chicago, IL, November 14-16. Wattanaphansak, S, C. Gebhart, M. Olin, A. Nuntraprasert and J. Deen. 2004. Measurement of the viability of Lawsonia intracellularis. Proc. 85th Conf. Res. Workers An. Dis., Chicago, IL, November 14-16. Beckler, D.C., N.L. Weber and C.J. Gebhart. 2005. Typing of Lawsonia intracellularis isolates by analysis of variable number tandem repeat profiles. Proc. Am. Assoc. Swine Pract., Toronto, Canada Wattanaphansak, S., T. Asawakarn, J. Deen, and C. Gebhart. 2005. Development of an enzyme-linked immunosorbent assay for the diagnosis of porcine proliferative enteropathy. Proc. Allen D. Leman Swine Conf. vol. 32, St. Paul, MN Asawakarn, T., A. Nuntaprasert, K. Kaur, V. Kapur, S. Wattanaphansak, and C. Gebhart. 2005. Expression of recombinant Lawsonia intracellularis proteins. Proc. Allen D. Leman Swine Conf. vol. 32. , St. Paul, MN Wattanaphansak, S., T. Asawakarn, J. Deen, and C. Gebhart. 2005. Development of an enzyme-linked immunosorbent assay for the diagnosis of porcine proliferative enteropathy. Rushmore Conference, Sept. 29-Oct.2, Rapid City, SD Asawakarn, T., A. Nuntaprasert, K. Kaur, V. Kapur, S. Wattanaphansak, and C. Gebhart. 2005. Expression and immunogenicity of proteins encoded by the genome of Lawsonia intracellularis. Rushmore Conference, Sept. 29-Oct.2, Rapid City, SD NE Extension Reports: Peterson, R. E., D. R. Smith, R. A. Moxley, T. J. Klopfenstein, S. Hinkley, and G. E. Erickson. Large-scale clinical trial to evaluate an experimental Escherichia coli vaccine. 2006 Nebraska Beef Report. University of Nebraska Cooperative Extension . Peterson, R. E., D. R. Smith, R. A. Moxley, T. J. Klopfenstein, S. Hinkley, and G. E. Erickson. Vaccination for Escherichia coli O157:H7 in market ready feedlot cattle. 2006 Nebraska Beef Report. University of Nebraska Cooperative Extension. Journal Articles: Carvajal, A., M.L. De Arriba, H. Rodriguez, A.B. Vidal, G.E. Duhamel, and P. Rubio. 2005. Brachyspira hyodysenteriae is relatively more prevalent than B. pilosicoli among commercial pig farms with diarrhoea in Spain. Vet. Rec., in press. Dassanayake, R.P., M.A. Griep, and G.E. Duhamel. 2005. The cytolethal distending toxin B sub-unit of Helicobacter hepaticus is a Ca2+- and Mg2+-dependent neutral nuclease. FEMS Microbiol Lett 251:219-225. Dassanayake, R.P., G. Sarath, and G.E. Duhamel. 2005. Penicillin-binding proteins in the pathogenic intestinal spirochete Brachyspira pilosicoli. Antimicrob. Agents Chemother. 49: 1561-1563. Dassanayake, R.P., Y. Zhou, S. Hinkley, C.J. Stryker, G. Plauche, J.T. Borda, K. Sestak, and G.E. Duhamel. 2005. Characterization of cytolethal distending toxin of Campylobacter species isolated from captive macaque monkeys. J. Clin. Microbiol. 43:641-649. Smith, D. R., R. A. Moxley, S. L. Clowser, J. D. Folmer, S. Hinkley, G. E. Erickson, and T. J. Klopfenstein. 2005. Use of rope-devices to describe and explain the feedlot ecology of Salmonella by time and place. Foodborne Pathog. Dis. 2:61-69. Smith, D. R., R. A. Moxley, S. L. Clowser, J. D. Folmer, S. Hinkley, G. E. Erickson, and T. J. Klopfenstein. 2005. Use of rope-devices to describe and explain the feedlot ecology of Escherichia coli O157:H7 by time and place. Foodborne Pathog. Dis. 2:50-60. Shivaprasad, H.L., and G.E. Duhamel. 2005. Cecal spirochetosis caused by Brachyspira pilosicoli in commercial turkeys. Avian Dis., in press. Book Chapters: Hampson, D.J., and G.E. Duhamel. 2005. Porcine colonic spirochetosis/Intestinal spirochetosis. In: Diseases of Swine. 9th ed. Straw BE, DAllaire S, Mengeling WL, and Taylor DJ (eds). Iowa State University Press, Ames, Iowa, pp. 755-767. Other Presentations and Published Abstracts: Bretschneider, G., E. M. Berberov, and R. A. Moxley. 2005. Role of the Tir protein in Escherichia coli O157:H7 intestinal colonization of adult cattle. Nebraska Symposium on Interdisciplinary Graduate Science Research, University of Nebraska-Lincoln, Lincoln, NE, Sept. 27 (Poster). Bretschneider, G., E. M. Berberov, and R. A. Moxley. 2005. Role of the Tir protein in Escherichia coli O157:H7 intestinal colonization of adult cattle. Third International Rushmore Conference on Enteric Diseases, Rapid City, SD, Sept. 29-Oct. 1 (Poster). Dassanayake, R.P., C.J. Stryker, R.K. Johnson, C.J. Gebhart, K.W. Post, S. Hinkley, W.T. Muraoka, I.V. Wesley, and G.E. Duhamel. 2005. The US porcine Campylobacter coli are negative for cytolethal distending toxin activity. 85th Annual Meeting Conference Research Workers in Animal Diseases, St. Louis, Missouri, November 6-8, (Poster). Dassanayake, R.P., and G.E. Duhamel. 2005. The cytolethal distending toxin B sub-unit of Helicobacter hepaticus localizes to the nucleus and is sufficient for intoxication of eukaryotic cells. 3rd international Rushmore Conference on Enteric Diseases, Rapid City, South Dakota, September 29 - October 1, (Poster). Dassanayake, R.P., C.J. Stryker, R.K. Johnson, W.T. Muraoka, I.V. Wesley, and G.E. Duhamel. 2005. Characterization of a novel Campylobacter cytolethal distending toxin from Campylobacter hyointestinalis subsp. hyointestinalis isolated from humans and pigs. 3rd international Rushmore Conference on Enteric Diseases, Rapid City, South Dakota, September 29 - October 1, (Poster). Dassanayake, R.P., M.A. Griep, and G.E. Duhamel. 2005. The cytolethal distending toxin B subunit of Helicobacter hepaticus is a nuclear localizing Ca2+- and Mg2+-dependent endonuclease. 105th General Meeting of the American Society for Microbiology, Atlanta, Georgia, June 5-9, Abst. B-008, p. (Poster). Duhamel, G.E. 2005. Efficacy of antimicrobial agents for PCS control. Pig Progress, Enteric Diseases Special III., p. 6-8. Duhamel, G.E. 2005. Understanding of colitis in swine improved. Section 4 in Perspectives on Swine Disease Management, Novartis Animal Health, Basel, Switzerland, p. 1-6. Duhamel, G.E. 2005. In vitro and in vivo efficacy of antimicrobial agents for control of porcine colonic spirochaetosis. Section 5 in Perspectives on Swine Disease Management, Novartis Animal Health, Basel, Switzerland, p. 1-6. Erume, J., E. M. Berberov, and R. A. Moxley. 2005. Disruption of enterotoxin genes of enterotoxigenic Escherichia coli by allelic exchange using lambda Red-mediated recombineering. Nebraska Symposium on Interdisciplinary Graduate Science Research, University of Nebraska-Lincoln, Lincoln, NE, Sept. 27 (Oral). Erume, J., E. M. Berberov, and R. A. Moxley. 2005. Disruption of enterotoxin genes of enterotoxigenic Escherichia coli by allelic exchange using lambda Red-mediated recombineering. Third International Rushmore Conference on Enteric Diseases, Rapid City, SD, Sept. 29-Oct. 1 (Poster). Smith DR. 2005. The future of E. coli O157:H7 intervention in live cattle. Cardinal Meats Conference on Food Safety. Toronto, Ontario, Canada. June 24, 2005 (Oral) Smith DR. 2005. Can vaccination reduce the probability that feedlot cattle shed Escherichia coli O157:H7? Invited seminar, University of Guelph, Ontario Veterinary College. Guelph, Ontario, Canada. Apr 29, 2005 (Oral) Smith DR, Moxley RA, Klopfenstein TJ, Peterson RE, Erickson GE. 2005. Population-based strategies for monitoring food safety pathogens in feedlot cattle. Beef Industry Food Safety Council (BIFSCO). Orlando, FL. April 20, 2005. (Oral) Smith DR. 2005. Can vaccination reduce the probability that feedlot cattle shed Escherichia coli O157:H7? Beef Industry Food Safety Summit. Orlando, FL. April 19, 2005. (Oral) Smith DR. 2005. Multidrug-resistant Salmonella: the bovine practitioners role in public health. National Conference on Ground Beef Contaminated with Multidrug-Resistant Salmonella, Including S. Typhimurium DT104: An Emerging Public Health Concern. Tufts University School of Veterinary Medicine, Grafton, MA. March 7-8, 2005. (Oral) Smith DR. 2005. Preventing calf scours with the Sandhills Calving System. Cattlemens Seedstock Showcase. Phillipsburg, KS. Feb 7, 2005. (Oral) Smith DR. 2005. Preventing calf scours with the Sandhills Calving System. UNL Gudmundsen Sandhills Laboratory, Whitman, NE. Jan 4, 2005 (Oral) Smith DR. 2005. Preventing calf scours with the Sandhills Calving System. UNL Extension, Holt County, ONeill, NE. Feb 18, 2005. (Oral) Smith DR. 2005. Preventing calf scours with the Sandhills Calving System. UNL Extension, Brown, Rock, KeyaPaha Counties, Ainsworth, NE. Feb 17, 2005. (Oral) Smith DR. 2005. Preventing calf scours with the Sandhills Calving System. UNL Extension, Custer County. Broken Bow, NE Feb 3, 2005. (Oral) Smith DR. 2005. Preventing calf scours with the Sandhills Calving System. Montrose /Dell Rapids Veterinary Clinic Client Education Meeting, Dell Rapids, SD, Jan 27, 2005. (Oral) OH Book Chapters 1. Saif, L.J. 2005. Comparative biology of animal coronaviruses: Lessons for SARS. In: Severe Acute Respiratory Syndrome. (M. Peiris, et al ed), Blackwell Pub., Oxford, UK, pp. 84-99. 2. Saif, L.J. 2004. Animal Coronaviruses: Lessons for SARS. In: Learning from SARS: Preparing for the next disease outbreak. SARS workshop sponsored by the Institute of Medicine of the National Academy of Sciences, Washington, DC, Sept. 30-Oct. 1, 2003, pp 138-148. 3. Yuan, L., G. Stevenson and L.J. Saif. 2005. Rotavirus and Reovirus. In: Diseases of Swine. 9th Edition. Edited by Zimmerman, J. J. et al. Ames, Iowa: Iowa State University Press (in press). 4. Saif. L.J. and K. Sestak. 2005. TGEV and PRCV. In: Diseases of Swine. 9th Edition. Edited by Zimmerman, J. J. et al. Ames, Iowa: Iowa State University Press (in press). 5. Costantini, V. and L.J. Saif. 2006. Calicivirus and Rotaviruses. In: Fate and transport of zoonotic bacterial, viral and protozoan pathogens during swine manure storage, treatment, and land application, Council for Agricultural Science & Technology, NPB, (submitted). Refereed Journal Articles 1. Chang, K. O., S. S. Sosnovtsev, G. Belliot, Q. Wang, L. J. Saif, and K. Y. Green. 2005. Reverse genetics system for porcine enteric calicivirus, a prototype sapovirus in the Caliciviridae. J Virol 79:1409-16. 2. Wang, Q., M. G. Han, S. Cheetham, M. Souza, J. A. Funk, and L. J. Saif. 2005. Porcine noroviruses: genetic and antigenic relationships to human noroviruses. Emerg Infect Dis.(in press). 3. Wang, Q., M. K. Han, J. A. Funk, G. Bowman, and L. J. Saif. 2005. Genetic diversity and recombination of porcine sapoviruses. J Clin Microbiol. (in press). 4. Wang, Q., K. O. Chang, M. K. Han, S. Sreevatsan, and L. J. Saif. 2005. Development of a new microwell hybridization assay and an internal control RNA for the detection of porcine noroviruses and sapoviruses by reverse transcription-PCR. J Virol Methods. (in press). 5. Han, M.G., S. Cheetham, M. Azevedo, C, Thomas, and L. J. Saif. 2005. Immune responses to bovine norovirus-like particles with various adjuvants and analysis of protection in gnotobiotic calves. Vaccine. (in press). 6. Thomas, C., A. Hoet, S. Sreevatsan, T. Wittum, R. Briggs, G. Duff and L.J. Saif. 2005. Field transmission of bovine coronavirus and herd immunity against associated respiratory disease in feedlot cattle. AJVR (submitted). 7. Costantini, V., F. Loisy, L. Joens, F.S. LeGuyader and L.J. Saif. 2005. Human and animal enteric caliciviruses in oysters from different coastal regions of the U.S. Appl. Envir. Microbiol. (submitted). Abstracts 1. Wang, Q., M. G. Han, S. Cheetham, M. Souza, J. Funk and L. J. Saif. 2005. Detection of porcine noroviruses (PoNoV) from US swine and their genetic and antigenic relationships to human noroviruses (HuNoV). To be presented at the 86th Conference of Research Workers in Animal Diseases, St. Louis, Missouri, December 4-6, 2005. 2. Costantini, V., F. Loisy, J. Joens, F. LeGuyader and L.J. Saif. 2005. Update on calicivirus survey of U.S. market oysters: animal enteric calicivirus. 24th Annual Meeting of the Conference of American Society for Virology, University Park, Pennsylvania, U.S. Abst. #50-10. June 18-22, 2005. Thesis Thomas, C. 2005. Bovine enteric and respiratory viruses: Studies of bovine enteric calicivirus and bovine coronavirus. MS Thesis, The Ohio State University. SD Butler, J. E., D. Francis, J. Freeling, P. Weber, J. Sun, K. L. Nielsen and A. M. Krieg. 2005. Antibody repertoire development in fetal and neonatal piglets. IX. Three PAMPs act synergistically to allow germfree piglets to respond to TI-2 and TD antigens. J. Immunol. 175: 6772-6785. Boots, RE, DH Francis and J. Freeling. Factors Influencing F18+ Escherichia coli Receptor Expression in Weanling Pigs. Abstract. Third International Rushmore Conference on Enteric Diseases. Rapid City, SD. Christie A., V Sebastien, S Gibson, D Francis and V Brozel. 2005. Proteomic analysis of Escherichia coli O157:H7 growing in soil organic matter reveals a unique phenotype. Abstract. Third International Rushmore Conference on Enteric Diseases. Rapid City, SD. Francis D, A Erickson, R Moxley, W Zhang and E Berberov. 2005. Use of germ-free pigs in modeling enterotoxigenic E. coli infections. Invited Presentation. Third International Rushmore Conference on Enteric Diseases. Rapid City, SD. Kaushik R, S George, A Young, and D Francis. 2005. Lectin binding profile of porcine intestinal epithelial cell lines. Abstract. Conference of Research Workers in Animal Diseases, Annual Mtg. St Louis MO. Kaushik RS, S George, A Young, and D Francis. 2005. Lectin binding profile of porcine intestinal epithelial cell lines. Abstract. Third International Rushmore Conference on Enteric Diseases. Rapid City, SD. Koh S, S George, V Brözel, D Francis and R Kaushik. 2005. Porcine Intestinal Epithelial Cell Lines as an in vitro Model for Studying Pathogenesis of Enterotoxigenic Escherichia coli. Abstract. Conference of Research Workers in Animal Diseases, Annual Mtg. St Louis MO. Koh SY, S George, V Brozel, D Francis and RS Kaushik. 2005. Porcine intestinal epithelial cell lines as an in vitro model for studying pathogenesis of enterotoxigenic Escherichia coli. Abstract. Third International Rushmore Conference on Enteric Diseases. Rapid City, SD. Lindblom SJ, SJ Vilain, VS Brozel, RS Kaushik, S George, HH Stein, C Pedersen, D Francis, AJM Rosa. 2005. Characterization of microbial communities in weanling pigs using 16S rRNA gene sequences. Abstract. Third International Rushmore Conference on Enteric Diseases. Rapid City, SD. Zhang W, M Zhao, and D Francis. 2005. Mapping the Binding Domains of K88ac Fimbrial Adhesin. Abstract. Conference of Research Workers in Animal Diseases, Annual Mtg. St Louis MO. WA 1. Bae, W, KN Kaya, DD Hancock, DR Call, YH Park, and TE Besser. 2005. Prevalence and antimicrobial resistance of thermophilic Campylobacter sp. from cattle farms in the Northwestern United States. Applied and Environmental Microbiology 71:169-174. 2. Borucki, MK, J Reynolds, DR Call, T Ward, B Page, and J Kadushin. 2005. Suspension arrays for direct and high throughput subtyping of Listeria monocytogenes from genomic DNA. Journal of Clinical Microbiology 43:3255-3259. 3. Call, DR. 2005. Challenges and opportunities for pathogen detection using DNA microarrays. Critical Reviews in Microbiology 31:91-99. 4. Call, DR, MS Kang, J Daniels, and TE Besser. Accepted. Assessing genetic diversity in plasmids from Escherichia coli and Salmonella enterica using a mixed-plasmid microarray. Journal of Applied Microbiology. 5. Wright JG, Tengelsen LA, Smith KE, Bender JB, Frank RK, Grendon JH, Rice DH, Thiessen AM, Gilbertson CJ, Sivapalasingam S, Barrett TJ, Besser TE, Hancock DD,Angulo FJ. 2005. Multidrug-resistant Salmonella Typhimurium in four animal facilities. Emerg Infect Dis. 11(8):1235-41. 6. Besser TE, LeJeune JT, Rice DH, Berg J, Stilborn RP, Kaya KN, Bae W, Hancock DD. 2005. Increasing prevalence of Campylobacter jejuni in feedlot cattle through the feeding period. Appl. Environ Microbiol. In press. 7. Cobbold, R.N.; Rice, D.H.; Davis, M.A.; Besser, T.E.; Hancock, D.D. Long-term Persistence of Multi-drug resistant (MDR) Salmonella enterica Serovar Newport in Two Washington Dairy Herds. Journal of the American Veterinary Medical Association. In press. 8. Borucki, MK, CC Gay, J Reynolds, KL McElwain, SH Kim, DR Call, DP Knowles. Genetic diversity of Listeria monocytogenes strains from a highly contaminated dairy farm. Applied and Environmental Microbiology. In press.