Brief Summary of Minutes of Annual Meeting

Notes from NC1041 Meeting, Chicago, IL; December 56, 2009 (Submitted by Linda S. Mansfield,) Sunday Meeting 1. NC1041 Meeting Planning 2010 a. Leadership for next year i. Organizer serves a 2 year term as secretary and a 2 year term as chair ii. Linda Mansfield will write report and organize the meeting in 2010 and 2011. b. Tasks i. Prepare and agenda and assign section leaders 1. Rewrite should be on the agenda for the next year 2. Need to prepare outline for the proposal during the 2010 meeting 3. Set up an email list ii. Compile reports for meeting iii. Assemble writing team for beginning proposal&due date. c. The Next Proposal i. Impact documentation 1. short term and long term goals 2. It is useful to get our work into the popular press and to be able to document the public perception 3. Everyone in the group should fill out an appendix E at their AES unit offices ii. Timeline - by the next year put together a timeline 1. Rewrite should be on the agenda for the next year 2. We will by next year have feedback from the midyear review iii. Outline 1. We have some progress on the outline. 2. This task must be done during the next meeting 3. Bert Stromberg will bring format for the proposal rewrite to the next meetings iv. Define stakeholders and start getting input and support from them 1. Stakeholders reports will be very important in the rewrite 2. We must show that stakeholders have been involved 3. Who are our stakeholders? We need to know and demonstrate this. 4. MIMs system setup - Set up in NIMS system operated out of Maryland a. *Send information and ideas for renewal outline NIMS system b. We have to do a comparative study of what others are doing in this work. i. NC(North Central)1041 ii. Food Safety but not the same NCES1033 as Clemson Univ. 5. Stakeholders a. The Public b. Consumer groups c. Industry groups d. Pork Producers e. State Medical Epidemiologists f. WE NEED TO BE INTERESTED IN WHAT WE ARE DOING DESPITE THE FLUCTUATIONS OF THE STAKEHOLDERS v. Topic Discussions 1. Focus Areas a. Annual Priorities b. Intersections between agriculture and public health c. Veterinary medicine has a disconnect with Public Health d. Move to Food Safety thus public health animal models become important to us as a group e. We must distinguish ourselves from the NIFA group. We are not sure what will happen with this 2. Training the next generation of scientists a. Margarets work to train the next generation of agricultural scientists 3. International Work a. Why are you going over to Africa and South East Asia? i. This is done due to biosecurity and food safety that is an international issue ii. Name the diseases that can be moved around iii. Microbes that are translocated across international boundaries can be identified iv. Must justify to US institutions what the benefits are to the US partners for international actions b. Locations i. ILRE-international livestock institution ii. Walter Reed built a facility at Makerere University, Uganda for examining Avian Influenza iii. USAID in East Africa 4. Organisms on which to focus a. Interact with public health groups to use their lists of microorganisms of interest e.g. A, B and C list b. This task will help us in terms of identifying stakeholders c. Need to get basic info in this area 2. NC 1041 should have a joint meeting a. US Japan Malnutrition Conference i. Richard Guemant Past Chair: ii. John Clemens-Korea b. USDA National lab-Qijang had this for an idea for joint collaborations c. 4th Rushmore Conference  we said we would have a meeting 3. Funding Issues a. How Hatch funds are designed to work i. Hatch ==> divided into regional associations ==> there is a dollar allocation by a formula ==> 25% must be spent on multistate projects ==> institutions divide it up ==> no formula for these divisions ==> very little accountability ii. Very few of us get funding from the AES for research iii. Very little of the funds are actually going for research iv. This is an administrative issue v. All institutions do this differently 1. Univ. Wisconsin has a competition vi. Formula funds and multistate funds 1. Each has different uses. 2. Multistate is more competitive vii. Financial data for group is backing in database. This needs to be updated. Use it for lobbying viii. In 2008 there was a bump in Hatch funds. 150 million more into Hatch 1. 245,000 2. 559,000 multidata 3. Reported through 547,000 by NIH 4. AES Stations 3X10^6 total state and 5. Hatch and Multistate funding records are managed through the Cris system 6. There is supposed to be in 1 to 1 relationship between money allocated and the work done; the reality doesnt match this. ix. Institute of Food and Agriculture. 1. Growth in scale of projects 2. AFRI funding was bumped up 3. Food Safety, international food security 4. Next Steps a. Compile the meeting notes and send to Dick b. Send a copy of last proposal to members. i. Tom Besser has this ii. NIMS has this c. Get a list of participants in 2009 i. Dick will add new people into the system ii. We have to fill out appendix E first d. Dave Francis is organizing the poster-talk judging for students. Does this transition? i. Piggyback with them e. We want to get graduate students involved i. Make list of trained grad students and what they are doing now 1. Grad students and postdocs should be invited a. Possibly one per station. b. We can forgo the registration fee for them. c. Money can be achieved through USDA as a separate grant ii. Junior faculty members can be encouraged to join. 1. Benefit is establishing a network of collaborators 2. Junior faculty need to have some incentive for joining 3. Exchange of reagents and collaborations are the benefits 4. Holding this meeting in conjunction with CRWAD made the cost reasonable. This is not likely to change f. Meeting Ideas i. NRSP8-genomic support project ½ day invited presentations ii. 2nd ½ day-station reports iii. All room  mostly plant and animal genomics iv. Symposia 1. could be organized on common topics and start, then-have discussions by breakouts 2. Symposium session becomes part of the meeting 3. Speakers a. Tim Johnson b. Syed Hashsham  microbial ecology 4. Topics for a symposium on Sunday morning a. *Decision-to think about topics and send soon b. Then we will solicit some speaker names c. Gut microbiome d. Metagenomics - Andy Banson i. Pyrosequencing ii. Illumina sequencing 5. Funding a. Try to get money from Bob b. Raise dues slightly to find this c. Corporate sponsors might pay for this i. Can we get a corporate sponsor? g. Midterm Progress Report is due soon i. Seek collaborations that gain funding ii. Technology transfer iii. Funding levels are always in debate iv. Rewrite is due in 2011 v. What is the status and future of the Rushmore Conference? vi. SAES-funding report. vii. NIFA opportunities 1. Mark Robinson is the NIFA liaison 2. Need participation of group members viii. Nar Agriculture+ &. Org. funding issues NC1041

Objective 1. Focus on emerging diseases- Identify, characterize and develop improved detection methods related to newly recognized, novel or emerging causes of zoonotic enteric disease and enteric pathogens of cattle and swine. A. S. enterica Arizona 1) Sampling oysters for Salmonella from the Yaquina river. Oysters were examined from January to November with 26 positive for Salmonella giving us a 2.9% prevalence. Oysters were only positive for Salmonella during the spring and summer months. Of the Salmonella isolates collected, 34% (9/26) were S. Newport. 2) Identification of colonizing/invasion genes. The whole genome of S. Newport genotype JJPX01.0014 was sequenced. The contigs aligned to NC_011080 with greater than 99% homology. One large difference was the presence of a 30,000 bp insertion in NC_011080 that is not present in S. Newport JJPX01.0014. Kansas 1) Norepinephrine Increases Horizontal Gene Transfer Rates Between Enteric Bacteria. The effect of catecholamines on the rate of conjugative transfer between enteric pathogens was assessed. There was a significant increase in the rate of horizontal gene transfer of a conjugative plasmid between Salmonella and E. coli in the presence of norepinephrine. Minnesota Cultivation medium affects the accuracy of S. enterica surveillance. Four different Salmonella strains were competed in broth or bovine feces at varied combinations and concentrations. In all experiments, S. Newport was the most competitive, regardless of the starting concentration and cultivation protocol. One strain of S. Typhimurium was rarely detected in competition, even when it was the only strain present in bovine feces. Overall, the probability of detecting a specific Salmonella strain had little to do with its starting concentration in the sample. North Dakota 1) Shedding of Salmonella in beef cattle at different production stages. Prevalence of Salmonella in calves remained constant (~50%) September & November 2008, peaked in December 2008 (100%) and then was dropped (27.7%) in February 2009. It rose again in June to 48% at slaughter. AMR patterns remained consistent throughout the study for cattle with resistance observed against the same six antimicrobials. During February 2009 some calves had isolates resistant to cefoxitin. A high association of the presence of the integrase gene was observed in cattle and calves. A negative association was seen towards Cefoxitin and the presence of the integrase. 2) Characterization of clinical Salmonella isolates from cattle and humans in the US and Uganda: AMR and class 1 integron. Pan susceptible isolates were 14.3% in cattle and 19.3% in humans. For cattle and human isolates 64% and 25%, respectively, were resistant to 2 or more antibiotics while 48% and 11%, respectively, had resistance to e5 antimicrobials. Among isolates from US 25% of the cattle samples were positive for for class 1 integron. Among the 74 isolates from Uganda all 100% were resistant to e5 antimicrobials. Relatively high resistance to ciprofloxacin was seen in 66.7% cattle and 64% humans. A total of 45.8% of human and 46.2% of cattle isolates tested positive for presence of integron 1. B. C. jejuni Arizona 1) Feedlot Sampling for the Presence of C. jejuni. Half of the cohort (18) received a natural diet and the other half a similar diet with rumensin, tylosin and implants of estradiol and progesterone. Flies and pigeons were positive for C. jejuni at arrival of the calves. Later, pen floors, feed bunks, and the watering units were positive. During the first three months 2 of 36 calves were positive for C. jejuni. However, the next six months all samples showed increases in the presence of C. jejuni except feed and flies. On the day of processing all cattle fecal samples were positive. An increase in the percentage of carcasses was positive for C. jejuni after evisceration with the highest number of positives coming from the ventral midline. Carcass samples and ground beef samples from the carcasses were negative. 2) Sequencing and annotation of C. jejuni strains S3 (poultry) and M129 (clinical). C. jejuni M129 genome resulted in 11 large contigs at 42x coverage and a genome size of 1.61Mb. Previous studies have demonstrated the presence of four genomic islands (CJIE1, 790bp; CJIE2, 1,109bp; CJIE3, 644bp; and CJIE4, 953bp) encoded by the fluid producing strain RM1221 that are present in or have limited distribution in other C. jejuni strains. Complete RM1221 islands 1, 2 and 4 are present in the genomes of fluid producing strains S3; none of these islands are present in the invasive non-fluid producing strain M129. 3) Identification of C. jejuni proteins in intestinal fluid exudate of infected piglets. One ml of filtered pooled intestinal fluid from pigs infected with RM1221 or filtered pooled fecal material were pooled and subjected to 2D gel electrophoresis. Four unique proteins were detected: CJE0556, CJE0595, CJE1447, and CJE1464. These proteins were found to be encoded within genomic islands CJIE2 (CJE0556, CJE0595) and CJIE4 (CJE1447, CJE1464). Michigan C57BL/6 interleukin-10-/- mice were infected with seven genetically distinct C. jejuni strains. Four strains colonized the mice and caused disease; one colonized with no disease; two did not colonize. A microarray comparison of the strain that colonized mice without disease to C. jejuni 11168 that caused disease revealed that putative virulence determinants, including loci encoding surface structures known to be involved in C. jejuni pathogenesis and differed from or were absent in the strain that did not cause disease. Ohio Nearly 60% of cattle populations carry Campylobacter. Occupational exposure to raw meat, consuming ground beef, drinking unpasteurized milk and contact with cattle are implicated in Campylobacter infections. Campylobacter spp were detected in 24% of fecal samples. 71 C. jejuni isolates, 130 C. coli isolates, and 63 other spp of Campylobacter were isolated and confirmed by multiplex. C. L. intracelluaris Minnesota 1) Involvement of cell-mediated immune response and specific local mucosal IgA production in L. intracellularis experimental infection. Five-week-old pigs were inoculated with L. intracellularis, intestinal mucosa homogenate from proliferative enteropathy diseased pigs, or mock infection solution. Weak IFN-³ production was detected in one pig of the pure culture group and two pigs of the mucosal homogenate group 14 days pi and in two animals of each group 20 days pi. All pigs in both inoculated groups were seropositive for Lawsonia (IgG) on day 20. Inoculated pigs showed very weak dose dependent DTH reactions. Eight pigs from the pure culture group and seven from the mucosa homogenate group had detectable IgA titers in the intestinal lavage 22 days pi. Ohio Detection and quantification of L. intracellularis. A SYBR green quantitative PCR assay targeting a unique hypothetical protein was developed for detecting and quantifying L. intracellularis. The method detects as few as 3 copies per PCR reaction of the bacterium growing in IEC-18 rat epithelial cells. The qPCR assay was successful in detecting L. intracellularis in fecal samples collected from pigs. D. E. coli Kansas 1) A Multiplex PCR procedure for Detection of Six Major Virulence Genes in E.coli O157:H7. A multiplex PCR procedure that detected six major virulence genes, fliC, stx1, stx2, eae, rfbE, and hlyA, in E. coli O157:H7 was developed. The procedure was validated with a total of 222 isolates. Fecal samples spiked with E. coli O157 amplified all six genes if the concentration of E. coli O157 was 104 CFU/g of feces. However, if PCR was carried out after 6 h enrichment, the detection limit was 10 CFU/g of feces. 2) Genetic Relatedness of E. coli O157 Isolates from Cattle Feces and Beef Carcasses. We used PFGE to characterize E. coli O157 isolates from pre-evisceration carcasses and feces that were recovered from 37 E. coli O157-positive truckloads at a commercial abattoir. Among all isolates, there were 17 PFGE types (95% homology) and 37 subtypes (100% homology). Specific subtypes were detected on multiple occasions and from different sample types within loads, among loads, and among days. Within truckload, the percentages of carcass isolates that were identical to high-shedder or low-shedder fecal isolates, as determined by PFGE, were 69.2% and 46.0%, respectively. The percentages of carcass isolates that were the same subtype as high-shedder or low-shedder fecal isolates were 35.3% and 58.8%, respectively. 3) Genetic Variations in Shiga Toxin-producing Abilities of Bovine and Human E. coli O157:H7. Reverse passive latex agglutination test was used to evaluate 107 isolates (50 human, 57 bovine) for Stx1 and Stx2 production. Stx2 production of e1:8 was found in 86.0% of human isolates compared to 26.3% of bovine isolates. Bovine isolates with the presence of the TNP regions were associated with significantly lower Stx2 production, while the Q933 gene was associated with higher Stx2 production. Q933 was a better indicator of high Stx2 production by human and bovine isolates and may be a useful screening method to assess their potential to cause human disease. 4) Effects of Mucin and its Carbohydrate constituents on E. coli O157 Growth in Batch Culture Fermentations with Ruminal or Fecal Microbial Inoculums. Our objective was to test the effects of mucin and its carbohydrate constituents on in vitro growth of E. coli O157 in ruminal or fecal microbial fermentations. In ruminal fermentations, fucose, mannose, glucuronic acid, galacturonic acid, glucosamine, galactosamine, and mucin had no effect on E. coli O157 concentration compared to the control fermentation. At 24 h, the concentration of E. coli O157 in fermentations with galactose was lower than the control. However, including gluconic acid as substrate increased E. coli O157 concentration at 24 h. In fecal fermentations, mannose, galactose, gluconic acid, glucuronic acid, galacturonic acid, glucosamine, and mucin increased E. coli O157 growth compared to control at 24 h, while galactosamine and fucose did not. Gluconic acid was the most stimulatory substrate. South Dakota 1) Expression of heat-labile enterotoxin increases E. coli adhesion to intestinal epithelial cells. We used the K88ac adhesin producing E. coli strain 1836-2: 8035 containing the intact LT operon cloned into pBR322; 8221 containing the LT operon a mutation in the A subunit; and 8589 containing only the cloned LTb subunit. We found that the strains expressing the intact toxin, a non-toxigenic mutant of LT or the toxins B subunit (8035, 8221, 8589) each were able to induce significantly higher adhesion when compared to a non-toxigenic, adhesin expressing, or non-ETEC strains (8017, G58-1 respectively). 2) Avirulent enterotoxigenic E. coli strains act as probiotics against pathogenic K88+ E. coli. We utilized a piglet ETEC challenge model to test the ability of non-pathogenic ETEC constructed strains to colonize the intestine and competitively exclude virulent ETEC. All K88ac receptor-positive piglets in the placebo group developed diarrhea and became dehydrated after 12 hours. Piglets inoculated with either strain containing the modified LT (8221 or 8488) did not exhibit clinical signs of disease following ETEC challenge while piglets inoculated with 8017 showed mild to no diarrhea. This study suggests that pre-inoculation with avirulent strains expressing adhesive fimbriae and a non-toxic form of LT provides significant protection from ETEC challenge when provided 24 hr before that challenge. 3) Assessment of Efficacy of oral E. coli constructs for protecting weaned pigs from enterotoxigenic E. coli. Piglets suckled their dams for 5 days then weaned to milk replacer. they were inoculated orally with a placebo or with isogenic E.coli strains at 7 days of age and again at days 14. At 21 days, piglets were challenged with the wild type virulent ETEC strain 3030-2 (K88+/LT/STb). Following challenge, all receptor positive pigs in the placebo control group developed diarrhea and became dehydrated. None of the piglets inoculated with any of the isogenic strains exhibited any clinical signs of disease following ETEC challenge. E. Calicivirus Ohio 1) Attachment and inactivation of foodborne enteric caliciviruses in lettuce. We assessed attachment of enteric caliciviruses to lettuce by immunofluorescence, infectivity assays and real time RT-PCR. For post-harvest processing, the efficacy of different treatments on reduction of infectious enteric caliciviruses in vegetables was evaluated. We conclude that at the virus pI, virions may precipitate to form aggregates that may facilitate virus binding to lettuce. The overall virus titers did not change after incubation at RT for 1 hr (pH 4-8); < 1.0 log10 reduction occurred at pH 3. Three different elution buffers were tried to elute TC-Po/SaV from experimentally contaminated lettuce. Only minimum essential medium (MEM) plus 2% fetal bovine serum (FBS) (pH7.4 - 7.7) were positive. Thus TC-Po/SaV remains infectious on lettuce leaves after storage at 4°C for 1 wk, which is the general shelf life of lettuce. 2) Detect and characterize NoVs and SaVs from swine and determine their genetic and antigenic relationships to the human strains. We initiated a study to determine the relationship between NoV prevalence in barns, farms and production systems. To date, porcine NoVs were detected from 33% pooled fecal samples. No human-like NoVs have been found by using separate RT-PCR assays with human NoV-specific primers or calicivirus universal primers followed. Recently, we detected a new calicivirus (WGP93C strain) by using RT-PCR with the calicivirus universal primer set followed by sequencing analysis of the RT-PCR products. Nucleotide BLAST search based on virus-specific 265 nucleotides revealed 88% identity with the new calicivirus, St-Valerien-like virus. Objective 2. Focus on effective interventions- Develop and improve interventions and preventative measures to reduce the incidence and prevalence of infections of cattle and swine with enteric and food borne disease agents A. C. jejuni Arizona Effect of the Cj1534 mutation on the ability of C. jejuni to adhere to and invade epithelial cells. Cj1534, a putative bacterioferritin gene of C. jejuni, is up-regulated greater than 5-fold in studies conducted in chicks and in piglets. Mutation of the gene demonstrated a significant reduction in the colonization of chicks by the bacterium. Our current studies also demonstrated a reduction in colonization and invasion of epithelial cells in vitro by a Cj1534 mutant. Michigan C. jejuni is linked to the development of autoimmune diseases. We investigated the role of TLR2, TLR4, MyD88, and TRIF signaling in C. jejuni-induced inflammatory activation of DCs. DC upregulation of MHC-II and costimulatory molecules after C. jejuni challenge was profoundly impaired by TLR2, TLR4, MyD88, and TRIF deficiencies. Similarly, C. jejuni-induced secretion of IL-12, IL-6, and TNF-± was significantly inhibited in TLR2-/-, TLR4-/-, MyD88-/-, and TRIF-/- DCs compared to wild-type DCs; however, the magnitude of inhibition was greater in MyD88-/-, TRIF-/-, and TLR4-/- DCs than in TLR2-/- DCs. C. jejuni induced interferon regulatory factor-3 (IRF-3) phosphorylation and IFN-² secretion by DCs in a TLR4-TRIF-dependent fashion, further demonstrating activation of this pathway. TLR2, TLR4, MyD88, and TRIF deficiencies all impaired Th1-priming ability of C. jejuni-infected DCs. Thus, our results show that cooperative signaling through TLR4-MyD88 and TLR4-TRIF axes represents a novel mechanism mediating C. jejuni-induced inflammatory responses of DCs. Ohio 1) C. jejuni colonization factors. The twin-arginine translocation (TAT) pathway represents an important virulence mechanism in many bacterial pathogens. The deletion of TAT significantly compromised the bacterial ability to tolerate stress responses including biofilm formation and cause persistent infection in chickens. 2) PolyPhosphate Kinases. Understanding the physiological and genetic properties that allow C. jejuni to survive and adapt to various stress conditions is crucial for therapeutic interventions. Polyphosphate kinase 1 is a key enzyme mediating the synthesis of polyphosphate, an essential molecule for survival, mediating stress responses, host colonization and virulence in many bacteria. Our findings demonstrate that C. jejuni ppk1 mutant was deficient in poly P accumulation and was associated with decreased ability to form viable but non-culturable cells under acid stress. The ppk1 mutant also showed decreased frequency of natural transformation and increased susceptibility to various antimicrobials. Furthermore, the ppk1 mutant was characterized by a dose-dependent deficiency in chicken colonization. B. S. enterica Arizona 1) Vaccination of chicks with a S. Typhimurium strain (x9088) containing a plasmid vector expressing the Cj1534 gene. Hatchlings were immunized orally with viable cells of S. Typhimurium x9088 (pYA3493-empty vector; pYA4495-1-vector with Cj1534) on days 3, 10, and 16. The vaccinated birds were challenged on day 26 with viable cells of C. jejuni strain NCTC11168. Birds vaccinated with the Cj1534 protein demonstrated a C. jejuni reduction of 3.5 logs after challenge exposure. Kansas Efficacy of the S. Newport SRP® Vaccine in Feedlot Cattle. Our objectives were to determine the effects of a commercially available S. Newport siderophore receptor and porin protein (SRP®). There were no differences between SRP® vaccinated and control cattle in fecal prevalence of Salmonella or any cattle health or performance parameters. Salmonella were recovered from all ten replicates (pairs) of pens, with cumulative prevalence ranging from 1.5% to 22%. Cumulative prevalence of fecal shedding was 10.2% and 10.9% for SRP® vaccinated and control cattle, respectively. Washington 1) Sources and transmission of multidrug resistant Salmonella that cause infection in humans. Analysis of PFGE and MLVA genotype data of human- and bovine strains suggests that S. Typhimurium isolates are more genetically diverse than Newport isolates and that multidrug resistance within that serotype is polyclonal. 2) Characterization of bacterial antimicrobial resistance using a validated DNA microarray. We developed a 203-probe oligonucleotide array targeting resistance and virulence genes of Gram-negative pathogens. 59 dairy herds were sampled to estimate the rate of introduction of new multidrug-resistant (MDR) S. enterica strains onto commercial dairy herds. The rate of new MDR Salmonella strain introduction was 0.9 per herd-year. The rates for the most commonly introduced MDR Salmonella serovars were 0.4/herd-year for Typhimurium, 1.2/herd-year for Newport, and 0.1/herd-year for Dublin. 56% had at least one new MDR Salmonella introduction during the study period. The number of new MDR Salmonella strains acquired by dairy herds ranged from zero to 8. 13 herds had a history of clinical salmonellosis. Among these 13 herds, 6 herds acquired new MDR Salmonella strains, although these strains were different than historical clinical strains. Recently, a new clade of S. Typhimurium, WA-TYP035/187, was reported in cattle and humans in the Pacific Northwest. The objective of this study was to describe a possible mechanism of acquisition of third generation cephalosporin resistance in this clade. Ceftazidime resistance increased steadily among WA-TYP035/187 isolates from 0% in 1999 to77.8% in 2006. 59 commercial dairy farms were sampled 7 times over 15 to 21 mo to determine the role of animal movement, including off-farm rearing of heifers, in the inter-herd transmission of MDR Salmonella spp. Logistic regression models indicated that off-farm heifer raising, including contract heifer raising where heifers commingle with cattle from other farms, and herd size per 100-animal increment were significantly associated with the introduction of new MDR Salmonella strains. The negative binomial regression similarly revealed that commingled, herd size per 100 animals, and a history of clinical salmonellosis diagnosed before the study were significantly associated with the number of new MDR Salmonella strains that were introduced. C. E. coli Kansas 1) Efficacy of E. coli O157:H7 SRP-based Vaccine in Feedlot Cattle. We evaluated the efficacy of an anti-E. coli O157 SRP-based vaccine in feedlot. 60 cattle were randomly allotted to 3 treatment groups: control, vaccinated with 2 ml or with 3 ml with vaccine. The vaccine at the 3 ml dose reduced the prevalence of E. coli O157 compared to the control (17.7% vs 33.7%). A similar numerical trend was observed with the 2 ml dose (29.1%), but differences were not statistically significant. Additionally, the 3 ml dose of SRP vaccine reduced the number of days cattle tested culture positive for E. coli O157, and the number of days cattle were identified as high-shedders compared to control. 2) Associations between E. coli O157:H7 in Feces and on Hides at Harvest and Contamination of Pre-evisceration Beef Carcasses. We found 38.5% of hides and 10.5% of carcasses positive for E. coli O157:H7. All truckload-level predictors significantly affected the probability of an E. coli O157:H7-positive carcass, including presence of a high shedder within the truckload, high (> 25%) within-truckload fecal prevalence, and high (> 50%) within-truckload hide prevalence; the only significant animal-level predictor was having a positive hide. 3) Prevalence of E. coli O157:H7 in Gut Contents of Beef Cattle at Slaughter. The prevalence of E. coli O157:H7 in the rumen, cecum, colon, and rectum were determined. Overall prevalence of E. coli O157:H7 in cattle sampled was 20.3%. E. coli O157:H7 in rectal contents was positively associated with presence in the rumen or colon but not in the cecum. The majority (79 to 90%) of isolates obtained within the same animal shared a common PFGE type. There was no significant difference between gut locations in reduction following acid challenge. 4) Feeding Dried or Wet Distillers Grains at Varying Inclusion Levels to Feedlot Cattle Affects the Fecal Prevalence of E. coli O157:H7. The inclusion of DG increased the prevalence of E. coli O157:H7 compared to the control cattle. The type of DG did not impact prevalence. Prevalence was not different between cattle fed diets with 0 or 20% DG, however, cattle fed 40% DG had significantly higher prevalence than either 0 or 20% DG. Similarly, the prevalence of super shedders was not different between cattle fed diets with WDG or DDG. Prevalence of super shedders in cattle fed 0 or 20% DG were not different, however, cattle fed 40% DG diets had a higher prevalence of high shedders than cattle fed either 0 or 20% DG. Nebraska Efficacy of a prebiotic galactooligosaccharide for reducing intestinal colonization by enteropathogenic E. coli and enterohemorrhagic E. coli O157:H7. Two litters of gnotobiotic piglets were used in a 2 X 2 factorial design with four treatment groups: GOS vs control with 2 challenge strains (EPEC strain E2348/69 or E. coli O157:H7 Shigatoxin-negative ATCC strain 43888). All inoculated piglets developed gross and microscopic intestinal lesions consistent with EPEC and E. coli O157:H7 infection. GOS reduced adherence of EPEC and EHEC in both the ileum and colon. However, the only statistically significant effect was a reduction of EHEC in the colons of piglets. Washington Bovine-biased lineages of EHEC O157 were non-virulent in the neonatal piglet model. Bovine-biased lineages were discovered to uniformly lack stx2, but rather carried stx1 and/or stx2c. These results suggest that approximately half of the isolates of O157 from the bovine reservoir have significantly less virulence than the rest. D. L. intracellularis Minnesota 1) In vitro antimicrobial activity against 10 North American and European L. intracellularis isolates. Our in vitro results showed that each L. intracellularis isolate had a different antimicrobial sensitivity pattern and these data can be utilized as an in vitro guideline for the further antimicrobial evaluation of field L. intracellularis isolates. 2) In vitro assessment of the effectiveness of powder disinfectant (Stalosan® F) against L. intracellularis using two different assays. Our results indicate that Stalosan F in both powder and suspension forms is able to inactivate over 99% of L. intracellularis after 30min of exposure. 3) Evaluation of in vitro bactericidal activity of commercial disinfectants against L. intracellularis. Results demonstrate that QAC, combinations of QACs with aldehydes, and oxidizing agents would perform well for inactivation of L intracellularis under field conditions. E. Antimicrobial resistance Washington Direct competition of plasmid bearing strains demonstrates that blaCMY-2 plasmids incur a measureable fitness cost in the face of plasmid-free competitors (3-12% loss per generation). Using a oligonucleotide microarray we showed that most genes on the plasmid backbone are quiescent in broth culture (log and stationary phases). In addition, all of the known antimicrobial resistance genes are expressed constitutively. Ceftiofur was not associated with any clear change in the number of blaCMY-2 expressing bacteria and conjugation rates between donor and recipient strains were not different from control groups. The objective of a clinical field trial was to determine the effect of colostrum supplementation of the milk replacer ration on morbidity, mortality, feed intake, and weight gain of preweaned calves. Calves receiving supplemental colostrum had less diarrhea and received fewer antimicrobial treatments than control and placebo calves. The results indicated that calf diarrhea was associated with low serum IgG levels and low-weight calves. Grain consumption and weight gain over the first 28 d of life were significantly greater in colostrum-supplemented calves compared with control calves. We determined the effect of raising pre-weaned dairy calves without antimicrobials in the milk. Newborn calves were allocated to one of 4 groups. Calves in the conventional therapy (CT) group were treated with sulfamethoxazole/trimethoprim, spectinomycin, penicillin and bismuth-pectin for diarrhea. The targeted therapy (TT) group included bismuth-pectin for diarrhea and antimicrobial treatment only in cases of fever or depressed attitude. Conventionally treated calves had 70% more diarrhea than TT calves and AB-milk calves had 31% more diarrhea than the NoAB-milk calves. The TT calves have a higher average daily gain at 28 days and consumed more grain compared to CT calves. We described the geographic, farm type and animal type factors associated with multiple antimicrobial resistance in bovine fecal E. coli. MAR (multiple antimicrobial resistance) was higher in isolates obtained from California cattle compared to Washington and Oregon cattle. MAR progressively decreased in isolates from calves on calf ranches, feedlot steers, dairy cattle and beef cow-calf cattle. MAR was higher in isolates obtained from calves than from adult cattle, in isolates from conventional farms than from organic farms, and in isolates from beef cow-calf farms in proximity to intensive dairy farm regions than in remote locations. F. Porcine Group A Rotavirus Ohio Rotavirus-like particle vaccines with/without attenuated rotavirus priming. We evaluated numbers of rotavirus-specific antibody-secreting cells (ASCs) and virus neutralizing (VN) antibody titers against Wa RV. The highest numbers of RV-specific IgA and IgG ASCs pre-challenge were induced in ileum by the AttMRV+2/6VLP vaccine, whereas the 2/6/7VLP+2/6/4VLP vaccine induced IgA and IgG ASCs in ileum only post-WaRV challenge. Both vaccines induced statistically similar levels of serum IgG antibodies pre-challenge, but the AttMRV+2/6VLP vaccine stimulated significantly higher serum IgA pre-challenge and serum IgG and IgA antibodies post-challenge. VN titers to the Wa strain were statistically higher pre-challenge for the AttMRV+2/6VLP vaccine than the 2/6/7VLP+2/6/4VLP vaccine but statistically similar post-challenge. Both vaccines induced similar low levels of heterotypic protection against diarrhea (36-37%) compared to 71% for the homotypic G1 AttWaRV+2/6VLP vaccine. G. Cornavirus Ohio Clarify the effects of experimental dual-infection by porcine reproductive and respiratory syndrome virus and porcine respiratory coronavirus on respiratory disease in pigs as a model for respiratory viral co-infections potentially relevant to the Severe Acute Respiratory Disease Syndrome. We investigated the effects of a prior and ongoing PRRSV infection, on PRCV co-infection and clinical disease and the pathologic and immunologic interactions of the two viruses, as a model for respiratory viral co-infections potentially relevant to SARS. The PRRSV/PRCV pigs had greater reductions in weight gains and more severe gross or histological pneumonic lesions compared to either single infection at PRCV PID 2 to 21. The PRRSV/PRCV dual virus-infected pigs had significantly suppressed innate immune responses, as evident by reduced IFN-± level in lung and blood. In addition, systemic NK cell-mediated cytotoxicity was significantly reduced in PRRSV only infected pigs. Upon co-infection with PRCV, there was a synergistic suppression of NK cell-mediated cytotoxicity. Co-infection by PRRSV and PRCV led to enhanced PRRSV replication in lung, a trend towards increased serum Th1 (IFN³ and IL-12), but decreased Th2 (IL-4) cytokine responses, and clinically exacerbated PRRSV pneumonia. H. C. parvum Illinois Experiments are under way to characterize the mechanism of inhibition of sporozoite adhesion. Suppressive subtractive hybridization experiments aimed at identifying specific sporozoite genes expressed in response to host cell attachment or exposure to the inhibitory lipid indicate these processes occur without the necessity for attachment- or lipid-induced differential gene expression. This lipid, introduced as part of different dietary regimens, also is being examined for its ability to reduce or eliminate bovine cryptosporidiosis in newborn calves. I. Microbiome Minnesota Alteration of the Pig Distal Gut Microbiota by Tylosin as an Antibiotic Growth Promoter. We hypothesized that the effects of AGPs are mediated by compositional changes of the pig gut microbiota. Fecal samples from tylosin treated and tylosin-non-treated pigs were collected 5 times at 3-week intervals. The sequences of the V3 hypervariable region of 16S rRNA were determined. The pig distal gut bacterial communities of both groups were dominated by Firmicutes and Bacterioidetes accounting for > 80% of total sequences, and showed highly diverse community structure (Shannon > 4.3, and Simpson 1-D > 0.9). Most of the sequences (> 95% of the total sequences) were shared by pigs in the two treatment groups. Components of the classes Actinobacteria, Clostridia, Fibrobacter, and Erysipelotrichi were different between the groups. Objective 3. Focus on disseminating knowledge- Provide training and continuing education opportunities and dissemination of information to students, producers, veterinarians and diagnostic laboratories. North Dakota North Dakota State University and Makerere University Uganda offered the joint course International Animal Production, Disease Surveillance and Public Health in summer 2009. Eight students from the US and two students from MAK attended the course. In 2009, a planning grant from USAID was obtained. As a result of that grant, three planning meetings were held in East Africa and in Fargo, ND to develop a 10-year strategic plan on capacity building in Integrated Management of Transboundary Animal Diseases & Zoonoses in East & Central Africa. Washington WSU Developed an extension program centered around calf health and calf science. The issue of antimicrobial resistance in food animal agriculture was addressed by conducting clinical trials to assess alternatives to antimicrobials in dairy calf-raising and developing outreach to three different audiences.

Publications: Arizona Law, B.F., S. M. Adriance, L.A. Joens. Comparison of in vitro Virulence Factors of Campylobacter jejuni to in vivo Lesion Production. Foodborne Pathogens and Disease. 6(3):377-85. 2009. Wilson, M. K., A. B. Lane, B. F. Law, W. G. Miller, L. A. Joens, M. E. Konkel, and B. A. White. Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulsed-Field Gel Electrophoresis, Multilocus Sequence Typing (MLST), and Repetitive Sequence Polymerase Chain Reaction (rep-PCR) Reveals Different Discriminatory Capabilities. Microbial Ecology 58:843855. 2009. DOI 10.1007/s00248-009-9571-3. Illinois Pineda, MF, L. L. Chan, Theresa B. Kuhlenschmidt, Mark S. Kuhlenschmidt, and Brian. T. Cunningham (2009) Rapid Label-Free Selective Detection ofPorcine Rotavirus using Photonic Crystal Biosensors for Groundwater Monitoring. IEEE Sensors Journal 9(4):470-477. Andres, A., Donovan, S.M. and Kuhlenschmidt, M.S. 2009. Isoflavones and virus infections (Invited Review), J. Nutritional Biochemistry (20(8):563-9). Liu, Y, Dao Janjaroen, Mark S. Kuhlenschmidt, Theresa B.Kuhlenschmidt, Thanh H Nguyen. 2009. Deposition of Cryptosporidium Parvum Oocysts on Natural Organic Matter Surfaces: Microscopic Evidence for Secondary Minimum Deposition in a Radial Stagnation Point Flow Cell. Langmuir (in press) Donovan, S.M., Andres, A., Mathai, R.A., Kuhlenschmidt, T.B. and Kuhlenschmidt, M.S. 2009. Soy formula and isoflavones and the developing intestine. Nutritional Reviews (In Press). McLaughlin, S.J., Kuhlenschmidt, T.B., Kalita, P.K. and Kuhlenschmidt, M.S. 2009. Interaction of cryptosporidium parvum oocysts with soil particles andvegetated filter strips (Submitted). Kansas Alam M.J., Renter D.G., Taylor E.V., Mina D.M., Moxley R.A., Smith D.R. Antimicrobial susceptibility profiles of Salmonella enterica serotypes recovered from pens of commercial feedlot cattle using different sampling approaches. Current Microbiology 2009; 58(4): 354-359. Rao S., Van Donkersgoed, J., Bohaychuk V., Besser T., Song X., Wagner B., Hancock D., Renter D.G., Dargatz D, Morley P.S. Antimicrobial drug use and antimicrobial resistance in enteric bacteria among cattle from Alberta feedlots. Foodborne Pathogens and Disease, in press. Van Donkersgoed, J., Bohaychuk V., Besser T., Song X., Wagner B., Hancock D., Renter D.G., Dargatz D. Occurrence of foodborne bacteria in Alberta feedlots. Canadian Veterinary Journal 2009; 50(2): 166172. B. Other Publications. Journal Articles: Coolon J.D., Jones K.L., Narayanan S., Wisely S.M. Microbial Ecology of the Intestinal Flora of P. maniculatus and P. leucopus of the Tri-State Mining District. Molecular Ecology, in press. Dodd C., Renter D.G., Fox J.T., Shi X., Sanderson M.W., Nagaraja T.G. Genetic relatedness of Escherichia coli O157 isolates from cattle feces and pre-intervention beef carcasses. Foodborne Pathogens and Disease, in press. Taylor E.V., Kastner J.J., Renter D.G. Challenges involved in the Salmonella Saintpaul outbreak and lessons learned. Journal of Public Health Management and Practice, in press. Walker, C., Shi X., Sanderson M. J., Sargeant J. S., Nagaraja T. G.. Prevalence of Escherichia coli O157:H7 in gut contents of beef cattle at slaughter. Foodborne Pathogens and Disease, in press. Reinstein S., Fox J.T., Shi X., Alam M.J., Renter D.G., Nagaraja T.G. Prevalence of Escherichia coli O157:H7 in organically and naturally raised beef cattle. Applied and Environmental Microbiology 2009; 75(16): 5421-5423. Alam M.J., Renter D.G., Ives S.E., Thomson D.U., Hollis L.C., Sanderson M.W., Nagaraja T.G. Potential associations between fecal shedding of Salmonella in feedlot cattle treated for apparent respiratory disease and subsequent adverse health outcomes. Veterinary Research 2009; 40:02. Jacob M.E., Fox J.T., Drouillard J.S., Renter D.G., Nagaraja T.G. Evaluation of feeding dried distillers grains with solubles and dry-rolled corn on the fecal prevalence of Escherichia coli O157:H7 and Salmonella spp. in cattle. Foodborne Pathogens and Disease 2009; 6(2): 145-153. Peterson G., Bai J., Narayanan S. A co-printed oligomer to enhance reliability of spotted microarrays. J Microbiol Methods 2009; 77(3): 261-266. Fox, J. T., Drouillard, J. S., and Nagaraja T. G. Competitive exclusion Escherichia coli cultures on E. coli O157 batch culture ruminal or fecal microbial fermentation. Foodborne Pathogens and Disease 2009; 6:193-199. Thomson D. U., Loneragan, G. H., Thornton A. B., Lechtenberg K. F., Emery D. A., Burkhardt D. T., Nagaraja T. G. .Use of a siderophore receptor and porin proteins-based vaccine to control the burden of Escherichia coli O157:H7 in feedlot cattle. Foodborne Pathogens and Disease 2009; 6:871-877. Fox, J. T., Thomson, D. U., Drouillard J. S., Thornton, A. B., Burkhardt D. T., Emery D. A., Nagaraja T. G. Efficacy of Escherichia coli O157:H7 siderophore receptor/porin proteins-based vaccine in feedlot cattle naturally shedding E. coli O157. Foodborne Pathogens and Disease 2009; 6:893-899. Fox, J. T., Drouillard J. S., Nagaraja T. G. 2009. Effects of mucin and its carbohydrate constituents on Escherichia coli O157 in batch culture fermentations with ruminal or fecal microbial inoculums. Journal of Animal Science 2009; 87:1304-1313. Thornton, A. B., Thomson D. U., Fox J. T., Loneragan G. H., Burkhardt D. T., Emery D. A., Nagaraja T G. .Siderophore receptor/porin proteins-based vaccination reduces prevalence and shedding of Escherichia coli O157:H7 in experimentally inoculated cattle. Journal of Food Protection 2009; 72:866-869. Jacob, M. E., Callaway T. R., Nagaraja T. G. Dietary interactions and interventions affecting Escherichia coli O157 colonization and shedding in cattle. Foodborne Pathogens and Disease 2009; 6:785-792. Theses and Dissertations: Taylor, Ethel. 2009. Foodborne disease in the United States: from surveillance to policy. MPH, Report. (Advisor, D.G. Renter) Lubbers, Brian. 2009. The Impact of Oxytetracycline Dosing on Bacterial Populations and Transfer of Resistance Elements between Salmonella and E. coli in Vitro and in Vivo. Ph.D. Dissertation. (Minor Advisor: S. Narayanan). Other Presentations and Published Abstracts: Renter, David G. (invited presentation). E. coli O157:H7  Dissemination and Persistence in Bovine Production Environments. The Third Governors Conference on Ensuring Food Safety: E. coli O157:H7  progress and challenges. University of Nebraska-Lincoln. 2009. Nagaraja, T. G. (invited presentation). Feeding Distillers Grains to Ruminants: What do we know so far? The Third Governors Conference on Ensuring Food Safety: E. coli O157:H7  progress and challenges. University of Nebraska-Lincoln. 2009. Renter D.G., Dodd, C.C., Thomson D.U., Nagaraja T.G. Salmonella in beef cattle production systems. BIFSCO - Beef Industry Safety Summit, San Diego CA. 2009. Dodd, C.C., Renter D.G., Thomson D.U., Nagaraja T.G. A randomized trial assessing effects of a commercially available Salmonella vaccine in commercial feedlot cattle. Conference for Research Workers in Animal Disease, Chicago, IL. 2009. #64. Dodd, C.C., Renter D.G., Nagaraja T.G., Shi X. Prevalence and persistence of Salmonella within pens of feedlot cattle. Conference for Research Workers in Animal Disease, Chicago, IL. 2009. #44. Peterson, G, Bai, J., Nagaraja, T.G., Patton, J., and Narayanan.S. Diagnostic Microarray for Human and Animal Bacterial Diseases. Conference for Research Workers in Animal Disease, Chicago, IL. 2009. Peterson, G., and Narayanan, S. Norepinephrine Increases Horizontal Gene Transfer Rates Between Enteric Bacteria. Conference for Research Workers in Animal Disease, Chicago, IL. 2009. Gart, E., and Narayanan, S. Influence of variable concentration of catecholamines on the growth of Citrobacter rodentium and expression of virulence genes. Conference for Research Workers in Animal Disease, Chicago, IL. 2009. Nagaraja, T. G. Pathogen concerns for organically-grown beef. Symposium on organic poultry and red meats-microbiology considerations: What do we know, what more do we need to know and how do we get there? Institute of Food Technology Annual Meeting, Anaheim, CA, 2009. Michigan Bell JA, St. Charles JL, Murphy AJ, Rathinam VA, Plovanich-Jones AE, Stanley EL, Wolf JE, Gettings JR, Whittam TS, Mansfield LS. 2009. Multiple factors interact to produce responses resembling spectrum of human disease in Campylobacter jejuni infected C57BL/6 IL-10(-/-) mice, BMC Microbiology, 2009 Mar 18;9:57, PMID: 19296832. Rathinam VA, Appledorn DM, Hoag KA, Amalfitano A, Mansfield LS. 2009. Campylobacter jejuni-induced activation of dendritic cells involves cooperative signaling through TLR4-MyD88 and TLR4-TRIF axes, Infection and Immunity, 2009 Mar 30, PMID: 19332531. Rathinam VA, Hoag KA, and Mansfield LS. 2008. Dendritic cells from C57BL/6 mice undergo activation and induce Th1-effector cell responses against Campylobacter jejuni. Microbes and Infection 2008 Oct;10(12-13):1316-24. Epub 2008 Aug 5, PMID: 18725315. Mansfield LS, Patterson JS, Fierro BR, Murphy AJ, Rathinam VA, Kopper JJ, Barbu NI, Onifade TJ, and JA Bell. 2008. Genetic background of IL-10-/- mice alters host-pathogen interactions with Campylobacter jejuni and influences disease phenotype, Microbial Pathogenesis 2008 Oct;45(4):241-57 Epub2008 Jun 11. Parthasarathy G and Mansfield LS. 2008. Recombinant IL-4 (rIL-4) enhances Campylobacter jejuni invasion of Intestinal Pig Epithelial Cells (IPEC-1), Microbial Pathogenesis, Accepted April 22, 2009. ISSN 0882-4010. Abstracts V.A.K. Rathinam, Daniel M. Appledorn, Kathleen A. Hoag, Andrea Amalfitano and Linda S. Mansfield. 2009. TLR4?MyD88 and TLR4?TRIF signaling axes cooperate in activating dendritic cells in response to an intracellular enteric pathogen. Pattern Recognition Molecules and Immune Sensors, Keystone Meetings, Banff, Alberta CA, March 2009. J.A. Bell, J.L. St. Charles, A.J. Murphy, V.A. Rathinam, A.E. Plovanich-Jones, E.L. Stanley, J.E. Wolf, J.R. Gettings, T.S. Whittam, Tristan Lefebure, Michael Stanhope, L.S. Mansfield. 2009. Pathogenicity of Campylobacter jejuni strains in C57BL/6 IL-10-/- mice varies with strain genetic background, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. J.P. Jerome, A.E. Plovanich-Jones, J.A. Bell, Linda S. Mansfield. 2009. Genetic changes during serial passage of Campylobacter jejuni in C57BL/6 IL-10-/- mice are associated with enhanced virulence, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. J.D. Olmstead, V.A.K. Rathinam, L.S. Mansfield. 2009. Determining organ specific recovery of Campylobacter jejuni in a murine disease model, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. JL St. Charles1,3, JA Bell1, and LS Mansfield. 2009. Development of murine models for study of Guillain-Barré Syndrome, FWDIRN Annual Meeting, Stevenson, WA. March 30-April 1, 2009. Minnesota BOOK CHAPTER Gebhart, C.J. and R.M.C. Guedes. (In press). Lawsonia intracellularis and the proliferative enteropathies. In C.L. Gyles, et al., (ed.), Pathogenesis of Bacterial Infections in Animals, 4th ed. Wiley-Blackwell Publishing, Hoboken, NJ. JOURNAL ARTICLES Wattanaphansak, S., R.S. Singer and C.J. Gebhart. 2009. Evaluation of in vitro bactericidal activity of commercial disinfectants against Lawsonia intracellularis. J. Swine Health and Production (accepted). Pusterla, N., R. Jackson, S. M. Mapes, J. Noland, R.M. Stenbom and C. Gebhart. 2009. Lawsonia intracellularis: Humoral immune response and fecal shedding in weanling foals following intra-rectal administration of frozen-thawed or lyophilized avirulent live vaccine. Vet. J. (in press). Guedes, R.M.C. and C.J. Gebhart. 2009. Evidence of involvement of cell-mediated immune response and specific local mucosal IgA production in Lawsonia intracellularis experimental infection. 2009. Can. J. Vet. Res. (in press). Pusterla, N. and C. Gebhart. 2009. Equine proliferative enteropathy caused by Lawsonia intracellularis. Equine Vet. J. (in press). Pusterla, N., J.R. Collier, S.M. Mapes, S. Wattanaphansak, C. Gebhart. 2009. Effect of administration of a virulent live vaccine of Lawsonia intracellularis on mares and foals. Vet Rec. 164:783-785. Wattanaphansak, S., R.S. Singer, R.E. Isaacson, J. Deen, B.R. Gramm and C.J. Gebhart. 2009. In vitro assessment of the effectiveness of powder disinfectant (Stalosan F) against Lawsonia intracellularis using two different assays. Vet. Microbiol. 136:403-407. Pusterla, N., R. Jackson, R. Wilson, J. Collier, S. Mapes and C. Gebhart. 2009. Temporal detection of Lawsonia intracellularis using serology and PCR in Thoroughbred horses residing on a farm endemic for equine proliferative enteropathy. Vet. Microbiol.136:173-176. Wattanaphansak, S., R.S. Singer and C.J. Gebhart. 2009. In vitro antimicrobial activity against 10 North American and European Lawsonia intracellularis isolates. Vet. Microbiol. 134:305-310. Wattanaphansak, S., Singer, R.S., Isaacson, R.E., Deen, J., Gramm, B.R., and Gebhart, C.J. In vitro assessment of the effectiveness of powder disinfectant (Stalosan F) against Lawsonia intracellularis using two different assays. Veterinary Microbiology 136: 403 (2009). Singer, R.S., Mayer, A.E., Hanson, T.E., and Isaacson, R.E. Do microbial interactions and cultivation media decrease the accuracy of Salmonella surveillance systems and outbreak investigations? Journal of Food Protection 72:707 (2009). Nebraska Refereed Journal Articles: Alam, M. J., D. Renter, E. Taylor, D. Mina, R. Moxley, and D. Smith. 2009. Antimicrobial susceptibility profiles of Salmonella enterica serotypes recovered from pens of commercial feedlot cattle using different types of composite samples. Current Microbiol. 58:354-359. B. Other Publications. Refereed Journal Articles: Smith, D. R., R. A. Moxley, R. E. Peterson, T. J. Klopfenstein, G. E. Erickson, G. Bretschneider, E. M. Berberov, and S. Clowser. 2009. A two-dose regimen of a vaccine against type III secreted proteins reduced Escherichia coli O157:H7 colonization of the terminal rectum in beef cattle in commercial feedlots. Foodborne Pathog. Dis. 6 (2):155-161. Moxley, R.A., D.R. Smith, M. Luebbe, G. E. Erickson, T. J. Klopfenstein, and D. Rogan. 2009. Escherichia coli O157:H7 vaccine dose-effect in feedlot cattle. Foodborne Pathog. Dis. 6 (7):879-884. Smith, D. R., R. A. Moxley, T. J. Klopfenstein, and G. E. Erickson. 2009. A randomized longitudinal trial to test the effect of regional vaccination within a cattle feedyard on Escherichia coli O157:H7 rectal colonization, fecal shedding, and hide contamination. Foodborne Pathog. Dis. 6 (7):885-892. Book Chapters: Moxley, R. A., D. R. Smith. 2010. Attaching-effacing Escherichia coli infections in cattle. Veterinary Clinics of North America: Food Animal Practice. (accepted) Extension Reports: Rich, A. R., A. N. Jepson, M. K. Luebbe, G. E. Erickson, T. K. Klopfenstein, D. R. Smith, and R. A. Moxley. 2009. Vaccination to reduce the prevalence of Escherichia coli O157:H7 in feedlot cattle fed wet distillers grains plus solubles. J. Anim. Sci. Vol. 87, E-Suppl. 3:268 Research Presentations and Published Abstracts: Klopfenstein, T. J., D. R. Smith, G. E. Erickson, and R. A. Moxley. Feeding distillers grains and E. coli O157:H7. 2009 Nebraska Beef Report. Agricultural Research Division, University of Nebraska Cooperative Extension, Institute of Agriculture and Natural Resources, University of Nebraska-Lincoln, MP92:47-49. Extension and Teaching Presentations: Smith, DR. Pre-harvest interventions for food safety. Universidad Autonoma Chapingo, Chapingo, Mexico, Mexico Date: 10/20/2009 Smith, DR. The ecology of food safety pathogens in live cattle: with emphasis on Escherichia coli O157:H7 and Salmonella. 1er Simposium Internacional de Inocuidad de alimentaria. Facultad de Medicina Veterinaria y Zootecnia. Universidad Nacional Autonoma d Mexico. Mexico City, DF, Mexico. Date: 10/21/2009 Smith, DR. On-farm practices that improve the safety of food: the example of Escherichia coli O157:H7 in cattle feedlots. 1er Simposium Internacional de Inocuidad de alimentaria. Facultad de Medicina Veterinaria y Zootecnia. Universidad Nacional Autonoma d Mexico. Mexico City, DF, Mexico. Date: 10/22/2009 Smith DR,Rich AR, Jepson AN, Luebbe MK, Erickson GE, Moxley RA, Klopfenstein TJ. A randomized trial to test the effect of vaccination and feeding high levels of wet distillers grains plus solubles on the probability for feedlot cattle to shed Escherichia coli O157:H7 in feces. International Society for Veterinary Epidemiology and Economics. Durban, South Africa. Date: 08/12/2009 Smith DR. Can what we feed cattle affect the safety of food? The effect of distillers grains on E. coli O157:H7 Date: 05/05/2009 DR Smith, AN Jepson, AR Rich, MK Luebbe, RA Moxley, GE Erickson, TJ Klopfenstein. A randomized trial to test the effect of vaccination and feeding high levels of wet distillers grains plus solubles on the probability for feedlot cattle to shed Escherichia coli O157:H7 in feces. Beef Industry Safety Summit. BIFSCO Date: 03/04/2009 Smith DR. An update on pre-harvest control of E. coli O157:H7. Nance County Cattlemen Assoc. Date: 02/16/2009 Smith DR. E. coli O157:H7 interventions and distillers grains. Nebraska Beef Feedlot Roundtable. Norfolk, Lexington, Bridgeport. Date: 02/10/2009 Smith DR. Beef cattle diseases: Can practitioners get paid for population thinking. 18th Annual Alabama Conference for Food Animal Veterinarians Date: 02/07/2009 Smith DR. Applying population thinking to neonatal calf diarrhea. 18th Annual Alabama Conference for Food Animal Veterinarians. Birmingham, AL Date: 02/07/2009 Smith DR. Population thinking in medicine. North American Veterinary Conference. Orlando, FL Date: 01/19/2009 Smith DR. The art and science of interpreting diagnostic tests. North American Veterinary Conference. Orlando, FL Date: 01/19/2009 Smith DR. The art and science of population diagnostics. North American Veterinary Conference. Orlando, FL Date: 01/19/2009 Smith DR. Applying population thinking to neonatal calf diarrhea. North American Veterinary Conference. Orlando, FL Date: 01/19/2009 Smith DR. Applying population diagnostics to the control of BVDV. North American Veterinary Conference. Orlando, FL Date: 01/19/2009 Smith DR. Applying population diagnostics to the control of Johnes disease. North American Veterinary Conference. Orlando, FL Date: 01/19/2009 AN Jepson, RA Moxley, DR Smith, GE Erickson, TJ Klopfenstein, D Rogan. Vaccine dose and serological response of feedlot cattle vaccinated against Escherichia coli O157:H7. Conference of Research Workers in Animal Disease. Chicago, IL Date: 12/07/2008 A. R. Rich, A. N. Jepson, M. K. Luebbe, G. E. Erickson, T. J. Klopfenstein, D. R. Smith, R. A. Moxley. E.coli O157:H7 -incidence, control, and distillers grain North Dakota Oloya, D. Doetkott, M.L. Khaitsa. 2009. Antimicrobial drug resistance and molecular characterization of Salmonella isolated from domestic animals, humans and meat products. J. Foodborne Pathogens & Disease, 2009, 6(3) 273-284. Luke C. Heider, M.L. Khaitsa, et al. 2009. Genetic and phenotypic characterization of the blaCMY gene from Escherichia coli and Salmonella enterica isolated from food-producing animals, humans, the environment and retail meat. J. Foodborne Pathogens & Disease, 2009, 6(3) 273-284. Papers submitted and currently under review: Ebot S.Tabe, James Oloya, Dawn K. Doetkott, Margaret L. Khaitsa. 2009. Characterization of multidrug-resistant Salmonella Typhimurium serovar Copenhagen isolated from feedlot cattle. Journal of Food Protection Trends, Manuscript ID FPT 09-29, Submitted September 29, 2009. Abstracts submitted to Conferences: Tabe ES, Oloya J, Doetkott DK, Bauer ML, Mahero MW, Khaitsa ML. 2009. Comparative effect of direct-fed-microbials on fecal shedding and genotypic diversity of Escherichia coli O157:H7 and Salmonella in Feedlot cattle. In Proceedings of the 12th International Symposium on Veterinary Epidemiology and Economics (ISVEE), August 10-14, 2009 Durban, South Africa. Michael Mahero, Susan Olet, Dawn Doetkott, Margaret Khaitsa. Characterisation of Antimicrobial Resistance (AMR) and Presence of Class 1 Integrons In Salmonella Serovars Isolated From Clinical Cases of Animals And Humans In North Dakota. North Dakota Academy of Science, April 30, 2009 Fargo ND, p.58. M.W.Mahero, D. K. Doetkott, S. Olet, D.K. Byarugaba, M. L.Khaitsa. Characterisation of Antimicrobial Resistance and Presence of Class 1 Integrons in Salmonella Isolated from Animals and Humans In North Dakota, USA and Kampala Uganda. North Dakota EPSCOR Conference, September 24, 2009 at North Dakota State University, Fargo ND. Poster #70. Nesemeier, B., Doetkott, D., Olet, S and Khaitsa, ML. Characterization of Salmonella spp. isolated from beef cattle from post weaning to slaughter. North Dakota EPSCOR Conference, September 24, 2009 at North Dakota State University, Fargo ND. Poster #71. Michael Mahero, D. K. Doetkott, S. Olet, D.K. Byarugaba, M. L.Khaitsa. Characterization of Antimicrobial Resistance and Presence of Class 1 Integrons in Salmonella Isolated from Animals and Humans In North Dakota, USA and Kampala Uganda. North Dakota. In Proceedings of the 69th annual meeting of the North Central Branch of the American Society for Microbiology, October 9-10, at La Crosse, WI. Ohio Azevedo, M.S.P., L. Yuan, A.M. Gonzalez, K. Jeong, T. Nguyen, C. Iosef, K. Lovgren-Bengtsson, B. Morein, and L.J. Saif. 2009. Protection and antibody secreting cell responses induced by a combined vaccine of attenuated human rotavirus oral or intranasal priming and intranasal 2/6 VP rotavirus-like particle boosting with ISCOM in gnotobiotic pigs. Clin. Vaccine. Immun. (submitted). Azevedo, M.S.P., A.M. Gonzalez, L. Yuan, K. Jeong, C. Iosef, T. van Nguyen, K. Lovgren-Bengtsson, B. Morein and L.J. Saif. 2009. Oral versus intranasal prime/boost regimen using attenuated HRV or 2/6 VLP with ISCOM influences protection and antibody secreting cell responses to rotavirus in neonatal gnotobio. Vaccine Immun. (in revision). D. Gangaiah, Issmat I. Kassem, Zhe Liu, and Gireesh Rajashekara. 2009. Importance of Polyphosphate Kinase 1 for Campylobacter jejuni: Viable-but-Nonculturable Cell Formation, Natural Transformation, and Antimicrobial Resistance. Applied Environ. Microbiol. (In Press) Drozd M., II. Kassem, W. Gebreyes, G. Rajashekara. 2010. A quantitative polymerase chain reaction assay for detection and quantification of Lawsonia intracellularis. J. Vet. Diagnos. Investigation (In Press) Rajashekara, G, Drozd, M., Gangaiah, D., Jeon, B., Liu, Z., and Zhang, Q., 2009, Functional Characterization of The Twin-arginine Translocation System in Campylobacter jejuni. Foodborne pathogens and Dis. 6: 935-945. Abstracts Saif, L.J. Gut reactions to rotavirus vaccines in a gnotobiotic piglet disease model: a priming lesson for neonatal mucosal immunity. The 5th Intl. Veterinary Vaccines and Diagnostics Conference, Madison, WI, July 19-23, 2009. Vlasova, A.N., M.S.P. Azevedo, H.R. Resque, A.M. Hartzler, and L.J. Saif. The effect of virulent human rotavirus challenge on TLR3 and 4 responses and plasmacytoid dendritic cell distribution in gnotobiotic pigs vaccinated with rotavirus-like particles (VLPs). Proc. 14th Intl. Congress of Mucosal Immunology (ICMI), Abstract, Boston, MA. July 5-9, 2009. Resque, H.R., A.N. Vlasova, M.S.P. Azevedo, A.M. Hartzler, and L.J. Saif. Rotavirus-like particle (CLP) vaccines with/without attenuated rotavirus priming induce heterotypic rotavirus antibody responses but low protection rates to heterotypic rotavirus challenge. Proc.14th Intl. Congress of Mucosal Immunology (ICMI), Abstract, Boston, MA. July 5-9, 2009. Wang, Q., Z. Zhang, and L.J. Saif. 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