Brief Summary of Minutes of Annual Meeting

1. The meeting was called to order by the Chair of NC-1202, Dr. Gireesh Rajashekara at 8:00 am on Dec 6. Dr. Rajashekara introduced Dr. Frank Blecha, the new NC-1202 Administrative Advisor from Kansas State University. 2. Administrative Advisor, Dr. Frank Blecha, gave a short presentation on specifics related to the completion of Midterm Review of NC-1202 multistate project, which was due on Dec 15th, 2014. 3. Nineteen (19) Progress Reports were presented from 9:00 am to 5:30 pm on Dec 6. 4. From 8:30 am to 10:00 am on Dec 7, Drs. Peter Johnson and Margo Holland provided an update on NIFA budget and funding, with introduction of two new programs (CARE and Exploratory) that began in FY2014 and may be included again in FY2015. The NC-1202 group expressed concerns and also provided suggestions for current NIFA funding opportunities, which include; 1) challenge and rationale to include three obligated components (Research, Extension, Education) for large integrated grants. Current policy presented a hurdle to package an effective and successful multi-state proposal; 2) possibility to include a new program/category for junior scientists and provide junior faculty greater opportunity. NIFA may adopt a NIH-like policy to provide incentives for New Investigators. Regarding support for new investigators, Drs. Peter Johnson and Margo Holland suggested NC1202 committee draft and send a recommendation to Dr. Sonny Ramaswamy (NIFA Director). 5. From 10:15 am to 12:00 pm on Dec 7, NC-1202 group members had vivid discussion on the following issues: • Progress Report. Midterm Review was reminded to be completed by 12/15/2014. Due to confidentiality concern for the date described in the station reports, it was suggested that the progress report will be only sent to the station representatives. • Budget. $75 registration fee was charged for 2014 NC-1202 annual meeting, which should cover all costs in 2014 with some leftover for 2015. It is expected that a lower registration fee will be charged for 2015 annual meeting. • Feasibility of organizing a symposium by NC-1202 group. The symposium (focused on enteric disease) will be submitted through CRWAD and is expected to attract more people and increase attendee numbers for CRWAD. Dr. Peter Johnson raised the issue of possible conflict with the symposium organized by other multistate project (e.g. PEDV focus). To address this concern, Dr. Frank Blecha suggested different groups should cooperate with each other. The symposium abstracts will be submitted through CRWAD. To select Keynote Speakers for the symposium, a 5-member committee (Linda Mansfield, Linda Saif, Jun Lin, Gireesh Rajashekara, Radhey Koushik) was established. • Student awards. NC-1202 offers awards for students to compete in the Pathobiology of Enteric and Foodborne Pathogens Section of CRWAD. This year three awards were offered, two for oral presentation and one for poster presentation. Drs. Radhey Kaushik (Chair) and Weiping Zhang (Vice-Chair) are in charge of the student award selection committee. Dr. Weiping Zhang will serve as Chair for the Pathobiology of Enteric and Foodborne Pathogens Section of CRWAD for 2015. The student awards were funded by the annual registration fee. To increase the number of abstracts submitted to the Pathobiology of Enteric and Foodborne Pathogens Section, the award size will be increased from 2015 with only one award ($200) for each category (Oral or Poster Presentation). • Collaboration and funding opportunities. Dr. Linda Saif suggested that NC-1202 submit a collaborative, integrated application to the 2015 NIFA Food Security Grant; the new RFA is expected to be released in early 2015. Specific topics suitable for this grant were discussed by the participants; ‘Microbiome & Gut Health’ was selected as the general theme for this application to address a significant and timely food security issue. Upon release of the 2015 RFA, a steering committee will be established to move the project forward. Dr. Mo Saif nominated Dr. Gireesh Rajashekara to lead the steering committee for this collaborative project in the near future.

Objective 1. Focus on emerging diseases: We will identify, characterize and develop improved detection and prevention methods related to newly recognized, novel or emerging causes of zoonotic enteric disease and enteric pathogens of food animals. A. Campylobacter jejuni Iowa Bacterial factors involved in an emerging SA clone that cause ruminant abortion: Comparative analysis and genetic work of C. jejuni strains showed capsule locus (CPS) and CJSA_0039 (putative tyrosine phosphorylated regulator) were absolutely required for induction of systemic infection and abortion in pregnant guinea pigs. The genes contributing to bacteremia were identified by Tn-seq analysis. A novel transformation strategy was utilized to determine the specific genetic determinants required for abortion. Immunogenicity of membrane-associated proteins of Campylobacter jejuni-associated with sheep abortion. We observed that CgpA, MetK, FabG had the strongest antigenicity, while HtrA, FlgL and Peb4 were less antigenic, and CJSA_0852 had only little reactivity with the sera tested. CgpA, HtrA and FlgL were chosen as subunit vaccine candidates to evaluate the protective immunity against bacterial challenge in our mice model of systemic infection and bacteremia. Results suggest that CgpA may be a potential subunit vaccine candidate against sheep abortion caused by C. jejuni. Prevalence of C. jejuni clone SA in feedlot cattle. A total of 3,184 fecal samples were collected from 66 different feedlot cattle herds in Iowa, Texas, Colorado, Missouri and Kansas, and cultured directly for Campylobacter. The overall prevalence rate of Campylobacter in cattle feces was 72% (2293/3184). Our findings indicate that Campylobacter is commonly prevalent in the U.S. feedlot cattle, and that clone SA constitutes a substantial portion of the commensal cattle C. jejuni population. Ohio Pathogenesis mechanisms associated with Campylobacter jejuni -induced abortion in ewes: We demonstrated that effectiveness of using sheep (the natural host) to study the pathogenesis of sheep abortion associated C. jejuni. We also highlighted the responses of the host and the progression of symptoms associated with infections in pregnant sheep. Control of broiler litter contamination with C. jejuni: We tested the impact of different chemical treatments [aluminum sulfate (Alu), sodium bisulfate (Sob), and magnesium sulfate (Mgs)] to reduce C. jejuni contamination of litter and cognate chicken colonization. Our findings demonstrate a strategy to control C. jejuni in litter, a valuable on-farm resource. Prevalence and antibiotic resistance properties of Campylobacter on Turkey farms: This study highlighted the high prevalence, genotypic diversity, and antimicrobial resistance of Campylobacter spp in commercial turkey from farm to slaughter. Michigan In 2013-14, we completed 13 experiments infecting mice with C. jejuni strains from patients with Guillain Barré Syndrome (GBS) and patients with colitis. We have developed several mouse models that will now permit study of C. jejuni strains with selected gene knockouts. We have developed a mouse model of autoimmune diseases arising after C. jejuni infection including Inflammatory Bowel Disease and Guillain Barré Syndrome that has yielded important new knowledge for the medical field and that has been published in a Nature Journal. We now know that the evolution of C. jejuni in the host can lead to changes in its surface coat that can trigger autoimmune disease in genetically susceptible hosts. Top accomplishments include 1) identifying at least three mouse models for studying GBS induced by C. jejuni, 2) Identifying neurological phenotypes and lesions in mouse models of GBS using a new digital gait analysis video camera and software, 3) demonstrating that colitis and GBS disease outcomes after C. jejuni infection have alternative T cell regulatory pathways in the C57BL/6 mouse model, and 4) showing effects of a Type III secretion system of C .jejuni on the outcome of infection with C. jejuni in an animal model. B. Salmonella Washington State Comparing antibiotic resistance in temporally related Salmonella from cattle and humans: We analyzed 5,124 human clinical isolates from Washington State Department of Health, 391 cattle clinical isolates from the Washington Animal Disease Diagnostic Laboratory, and 1864 nonclinical isolates from foodborne disease research on dairies in the Pacific Northwest. Salmonella Typhimurium (ST), Salmonella Newport (SN) and Salmonella Montevideo (SM) were the most common serovars in both humans and cattle. The ST and SN from cattle had greater probability of resistance to multiple classes of antimicrobials than ST and SN from humans (p<0.0001). However, occurrence of profiles unique to cattle and not observed in temporally related human isolates, indicates these profiles are circulating in cattle only. Further AMR diversity assessment using different measures and new techniques (e.g. bootstrapping) showed that although there were shared profiles between humans and cattle, the expected and observed number of profiles was different, suggesting Salmonella and associated resistance from humans and cattle may not be wholly derived from a common population. Most prevalent poultry-associated Salmonella serotypes (MPPSTs) differ in their susceptibility to widely used carcass sanitizer, chlorine: Significant proportion of marketed poultry meat in the US remains contaminated with few Most Prevalent Poultry-associated Salmonella seroTypes (MPPSTs). A simulation model-derived data suggest that MPPSTs differ in their susceptibility to chlorine and that the level of CME in immersion chilling is an important contributing factor in Salmonella survival. C. Shiga toxin-producing E. coli (STEC) Nebraska Detection of enterohemorrhagic Escherichia coli (EHEC) in samples of feces, hides, and carcasses of culled dairy cows at harvest. We determined the proportion of fecal, hide, and de-hided carcass surface samples positive for seven EHEC serotypes (O26, O45, O103, O111, O121, O145, and O157) in culled dairy cattle at slaughter using a convenience sampling approach. One hundred culled dairy cows from the western U.S. were sampled over 5 weeks at a commercial abattoir (June to July 2014). Our results suggest that adulterant EHEC may be highly prevalent in culled dairy cattle at slaughter, further emphasizing the importance of effective pre- and post-harvest interventions. Erosive colitis in a yearling feedlot heifer caused by enterohemorrhagic Escherichia coli O165:H25. An EHEC strain that was identified as serotype O165:H25 and positive for stx2c, eae-?, and ehxA was isolated from the tissue of erosive colitis in a yearling feedlot heifer. This is the first clinical case of EHEC O165:H25 infection identified in cattle, and also represents one of the oldest bovine animals known to be clinically affected with any EHEC serotype. Wyoming Development of MALDI-TOF MS protocols for diagnosis of antimicrobial resistant bacteria. A total of 1,177 presumptive AMR Escherichia coli (ciprofloxacin and cefotaxime resistant), Staphylococcus spp. (cefoxitin resistant), and Enterococcus spp. (erythromycin resistant) isolates from wildlife, feed, and water associated with concentrated animal feeding operations were analyzed using developed MALD-TOF MS method. Detailed analyses of mass spectral profiles identified numerous spectral peaks, which may be useful for subtyping applications, including differentiation of AMR phenotypes. Kansas Multiple STEC projects have been successfully completed, which include 1) summer and winter prevalence of STEC in feces of feedlot cattle. This study indicated that non-O157 STEC are rare, but generally follow the same seasonal pattern as O157 STEC. However, further research is needed to determine if this trend holds true across other cohorts of cattle, feedlots and regions of the country. 2) feedlot- and pen-level prevalence of 7 STEC serotypes in feces of commercial feedlot cattle. Results provided insight on the feedlot- and pen-level fecal prevalence of E. coli serogroups in pre-harvest commercial feedlot cattle. 3) Develop culture- and PCR-based methods to detect 7 major serogroups of STEC. This study refined culture-based methods and compare with PCR method for detection of six non-O157 STEC serogroups in cattle feces (n = 576). 3) Comparison of a four-plex real-time PCR assay, conventional PCR and culture-based method for detection of STEC in feces of feedlot cattle. Our results indicated the multiplex quantitative PCR (mqPCR) assay detected more samples as positive compared to conventional PCR or culture-based method; 4) Pooling of immunomagnetic separation beads does not affect sensitivity of detection of seven serogroups of STEC in cattle feces. Pooling of IMS beads is advantageous because of reduced time, labor and expense required to detect and isolate STEC-7.; 5) Application of multiplex real-time PCR for the detection of six major non-O157 E. coli serogroups in feces of feedlot cattle. The higher relative sensitivity coupled with reduction in time and labor make mqPCR assays a good alternative to conventional PCR and culture-based methods for detection of the six non-O157 E. coli serogroups in cattle feces; 6) Escherichia coli O104 in feedlot cattle feces: Prevalence, isolation and characterization. Cattle shed serogroup O104 in feces, but only a few strains (11.2 %) carried stx1 gene and none of the isolated strains carried genes characteristic of the hybrid pathotype; 7) Quantification of six non-O157 E. coli serogroups in cattle feces by spiral plating method. This study indicated that the spiral plating technique combined with PCR confirmation has the potential to be a useful technique to quantify non-O157 E. coli serogroups in cattle feces. D. Brachyspira hampsonii Minnesota Molecular characterization of emerging “Brachyspira hampsonii” isolates. The novel pathogen, “Brachyspira hampsonii”, consists of two distinct genetic clades (I and II) and the pathogenicity of each clade has been confirmed experimentally in pigs. With one exception, all evaluated swine systems were infected with a single specific clade. Sites and systems that were positive for “B. hampsonii” repeatedly over several years were found to be temporally infected with the same ST or two clonally related STs. E. Coronavirus Ohio Detection of porcine deltacoronavirus (PDCoV) in US swine, pathogenesis of OH strains in gnotobiotic (Gn) pigs. The PDCoV, a new coronavirus, has been simultaneously and frequently detected in diarrheic fecal samples from pigs. The original and Gn pig-passaged OH-FD22 and OH-FD100 samples were confirmed virulent and free of other viral pathogens. These PDCoV pools will provide a useful challenge strain for pathogenesis and vaccine studies. Our study, for the first time, confirmed that PDCoV induces clinical disease and pathologic lesions in pigs. Minnesota Rapid detection and phylogenetic analysis of porcine deltacoronavirus. A real time RT-PCR was developed to detect and characterize PDCoV. A majority of the positive PDCoV samples (n=69, 78%) were co-infected with at least one of the previously described pathogens while 20 (22%) were negative for previously described pathogens. The results indicate that PDCoV is a common pathogen in the Midwest. F. Rotavirus Minnesota Widespread rotavirus H (RVH) in commercially raised pigs. The swine RVH VP6 nucleotide percent identity had a higher similarity to the novel human RVH than swine RVB strains. In the United States, 30 positive RVH samples were detected form 10 different states. Sequencing analysis suggested RVH has been circulating in the United States swine population for at least one decade but probably longer. Detection and high occurrence of porcine rotavirus A, B, and C by real time RT-PCR in diagnostic samples. Qualitative porcine RVA, RVB, and RVC RT-PCR (RT-qPCR) assays were designed and 7508 porcine diarrheic samples were tested to estimate the percentage of RVA, RVB, and RVC over a period of approximately 2 years (from 2009 to 2011). Although RVA was detected at the highest overall percentage (62%), 33% and 53% of the samples were positive for RVB and RVC, respectively, indicating that both RVB and RVC are also epidemiologically important in the swine population. RVC was most predominant in young pigs (1–20 days of age), while RVA and RVB were most predominant in ?21 day old pigs. G. Antimicrobial Resistance Tennessee Regulation of beta-lactam resistance in Campylobacter jejuni. Our molecular and biochemical studies demonstrated that Cj0843c, a putative lytic transglycosylase involved in cell wall metabolism, was required for induction of ?-lactamase-mediated ?-lactam resistance in Campylobacter. The crystals of Cj0843c have been significantly improved (the crystal diffracted to 2.57 Å resolution) and will be used for structural analysis and subsequent molecular docking to identify specific inhibitors. Molecular basis of conjugation in Campylobacter jejuni. Conjugation is an important mechanism for horizontal gene transfer. Using C. jejuni as a model recipient strain, we have made significant progresses in the mechanisms of conjugation. First, we observed that heat shock treatment could dramatically increase conjugation efficiency of C. jejuni. Second, using a unique co-transformation strategy in conjunction with whole genome sequencing and comparative genomics analysis, we have identified factors required for high frequency conjugation in C. jejuni. Development of novel alternatives to antibiotic growth promoters (AGP) for enhanced animal health and food safety. Bile salt hydrolase (BSH), a widely distributed function of the gut microbiota, has a profound impact on host lipid metabolism and energy harvest. Our studies suggested that BSH inhibitors are promising alternatives to AGP for enhanced animal growth performance and food safety. The BSH from L. salivarius is an ideal candidate for screening desired BSH inhibitors. Ohio Prevalence and antibiotic resistance properties of Campylobacter on Layer farms: We have recently finalized a study that compared the antibiotic resistance properties of Campylobacter isolated from conventional and organic layer farms. We highlighted the role of layer farms as potential reservoirs for antibiotic resistance markers, which can pose a risk as potential environmental pollution. Our results also show that the type of farming can affect the emergence of antibiotic resistance in these settings. Notably, antibiotic resistance was detected even in isolates collected from farms under organic management. Kansas Effects of in-feed copper and tylosin supplementations on copper and antibiotics resistance in fecal enterococci of feedlot cattle. The objective was to investigate whether feed supplementation of copper, at elevated level, co-selects for macrolide resistance in fecal enterococci in the absence of tylosin. A longitudinal study indicated that due to the selection pressure feeding of elevated dietary copper and tylosin alone or in combination resulted in an increased prevalence of tcrB and erm(B) mediated copper and tylosin resistant fecal enterococci in feedlot cattle. Effects of in-feed copper, chlortetracycline, and tylosin on the prevalence of transferable copper resistance gene, tcrB, among fecal enterococci of weaned piglets. Longitudinal study found that the supplementation of antibiotics in combination with copper increased the prevalence of copper-resistant enterococci. Objective 2. Focus on preventions and interventions: We will develop and improve preventative measures and interventions to reduce the incidence and prevalence of infections of food animals with enteric and foodborne and waterborne pathogens. A. Campylobacter jejuni Iowa Molecular genetic analysis of C. jejuni strains. Toxin-antitoxin (TA) systems are pairs of genes that encode a stable toxin and an unstable antitoxin and their primary function is to maintain the plasmid by eliminating plasmid-free daughter cells through a post segregation killing mechanism. Our recent study first demonstrated that the existence of the type I and type II TA systems in pVir plasmid of C. jejuni. In this study, we report the identification and characterization of the type II toxin-antitoxin system in the pVir plasmid of C. jejuni clone SA strain IA3902, a highly abortifacient isolate from sheep. Our results establish that CJSA_pVir0046 and CJSA_pVir0046 encode a functional TA system in C. jejuni. Control of Campylobacter in poultry flocks. We conducted a longitudinal epidemiological study to identify the risk factors associated with variation in Campylobacter prevalence in U.S. commercial broiler production. Cecal contents (n=5521) and boot swabs (n=1029) were collected from 369 broiler flocks on 15 different farms with repeated sampling. In order to identify the risk factors associated with Campylobacter prevalence at the farm and house/flock levels, farms were divided into 2 categories based on the overall number of Campylobacter positive flocks reared during the entire study period (i.e., low prevalence vs high prevalence). Arizona C. jejuni vaccine development. Recombinant attenuated Salmonella vaccines (RASV) have been adapted to stably express protective antigens at high levels. Two replicates of heterologous challenge vaccination studies were conducted with both studies resulting in a 1 log reduction. A third replicate vaccine study testing dual doses (1ml each at ~1x1010 CFU) of vaccines expressing CjLAJ1 and CjLAJ2 and challenged with the homologous strain C. jejuni NCTC 11168 resulted in a 2-log reduction. Construction of a single vaccine expressing dual genes and optimization of current constructs is underway. Tennessee Development and evaluation of Campylobacter vaccine. The CmeC- and CfrA-based DNA vaccines have been modified for in ovo immunization. Two independent in ovo vaccination trials have been performed. The in ovo injection of DNA vaccines at 18th day of embryonation failed to trigger significant immune response in broilers and to confer protection against C. jejuni colonization. The chitosan encapsulated subunit mucosal vaccines have been prepared and will be evaluated in broilers in the future. Ohio The role of respiratory proteins in C. jejuni’s pathobiology: The respiratory protein methylmenaquinol:fumarate reductase (Mfr) contributed to the oxidative stress response. Mfr also affected the interaction of the bacterium with macrophages and chicken colonization. Formate dehydrogenase activity and localization are dependent on a functional twin arginine translocation system in the highly invasive strain C. jejuni 81-176. The role of inorganic polyphosphate (poly P) metabolism in C. jejuni’s pathobiology: Exopolyphosphatase/guanosine pentaphosphate phosphohydrolase (PPX/GPPA) also contributed to poly P and ppGpp homeostasis. The PPX/GPPA also modulated environmental stress responses and virulence in C. jejuni. In a separate study, we showed that poly P mediated the modulation of biofilm growth and stability in C. jejuni. Our findings further highlighted the importance of these molecules as potential targets for anti-C. jejuni therapeutics Michigan Janus-faced immunity to an enteric pathogen: How Campylobacter mediates colitis and autoimmunity. C. jejuni is a leading cause of foodborne enteritis and has also been linked to the autoimmune neuropathy, Guillain Barré syndrome (GBS). This study established the first mouse model of an autoimmune disease induced directly by an enteric bacterium. Innate immunity is insufficient to mediate severe C. jejuni induced colitis. Rag1-/- mice on the C57BL/6 background were challenged with C. jejuni alongside C57BL/6 wt mice that serve as a known positive colonization control. Our data showed that T cells are necessary for severe C. jejuni-induced colitis. C. jejuni colonization correlated with colitis induction in as much as the mouse genetic background that supports inflammation (IL-10-/-) had higher colonization extent by the end of the experiment than the mouse genotypes/treatments that did not (BL/6wt or Rag1-/- or Rag1-/- IL-10R blocked mice). Washington Evaluation of Passive Immunotherapeutic Efficacy of Hyperimmunized Egg-Yolk Powder (HEYP) Against Intestinal Colonization of Campylobacter jejuni in Chickens. The results showed that there were no differences in the cecal colonization of C. jejuni between HEYP treated and non-treated control chickens. The use of HEYP at the dose and the regimes used in current study is not efficacious in reducing C. jejuni colonization in chickens. B. STEC and ETEC Nebraska Study of the effects of glucose on heat-labile enterotoxin (LT) and heat-stable enterotoxin-b (STb) gene expression in porcine-origin enterotoxigenic E. coli (ETEC) using lux reporters. This study is critical to demonstrate regulation of the two enterotoxins (LT and STb) concurrently in a given strain and determine whether the effects occur in vivo in inoculated animals. In the past year, we constructed ?crp derivatives of Lux reporter strains, and also made CRP-complemented constructs. The expected effects of catabolite repression on the STb reporter were again seen in the CRP-complemented mutant strain and a trend toward catabolite de-repression was seen in the CRP-complemented LT reporter. Kansas Multiple ETEC projects have been successfully completed, which include 1) ETEC biofilm formation. Mutagenesis study demonstrated that biofilm formation was reduced in the fliC mutant but increased in the faeG mutant, as compared with the wild-type strain; 2) ETEC and intestinal cell autophagy. ETEC induced autophagy, as measured by both the increased punctae distribution of GFP-LC3 and the enhanced conversion of LC3-I to LC3-II. Inhibiting autophagy resulted in decreased survival of IPEC-1 cells infected with ETEC; 3) Role of methionine on intestinal cell autophagy during ETEC infection. Methionine deprivation resulted in enhanced ETEC adhesion and increased both the cytotoxicity and apoptotic responses of IPEC-1 cells infected with ETEC. 3) E. coli quorum sensing. The luxS deletion mutant exhibited changed flagella-related phenotypes. The mutant strain also displayed attenuated production of Stx2e. This study provides new information to the crucial function of luxS in regulating Shiga-like toxin producing E. coli virulence; 4) E. coli genome sequencing. The objective of this study was to report the draft genome sequence of enterotoxigenic Escherichia coli (ETEC) strain W25K, which causes diarrhea in piglets; 5) Bacterial virulence factors and host inflammation. The Escherichia coli NleH1 and NleH2 virulence proteins differentially regulate host transcription of innate immunity genes. NleH expression was detrimental to C. rodentium in C57BL/10ScNJ mice. C. rodentium that expressed both E. coli NleH1 and NleH2 was hypervirulent in C3H/HeJ mice; 6) ETEC and intestinal immunity. We found that ETEC infection promoted the production of pro-inflammatory cytokines. Meanwhile, ETEC infection affected sIgA transport and Paneth cell function. 7) Efficacy of a vaccine and a direct-fed microbial against fecal shedding of Escherichia coli O157:H7 and corresponding impacts on cattle performance in a commercial feedlot. A siderophore receptor and porin proteins-based vaccine (VAC) and a Lactobacillus acidophilus-based direct-fed microbial (DFM) were evaluated. We observed that the application of these two treatments differentially impacted both fecal shedding of E. coli O157:H7 and cattle performance outcomes, demonstrating need to consider potential food safety impacts as well as cattle and carcass performance, when evaluating potential costs and benefits of interventions; 8) prevalence of STEC-8 in muscoid flies in the confined cattle environment. Prevalence of STEC-8 in house flies has been assessed on the temporal and spatial basis. Washington We completed a comparative study involving US, Australia and Argentina strains of E. coli O157:H7 isolated from cattle, using SNP and SBI genotyping. Results strongly support for phylogeographic structuring. Countries with high incidence of human O157-related disease have much higher frequency of strains encoding Stx2a. Novel SNP lineages of E. coli O157:H7 comprised substantial proportions of the populations in countries outside of the US, where the SNP typing panel was developed. We participated in a second comparative study of geographic divergence of E. coli O157:H7 between the US and New Zealand. We conducted an oral immunization trial of cattle with E. coli O157:H7, using a pool of three spontaneously Stx-negative E. coli from the EHEC1 lineage We are continuing to test the hypothesis that high summertime seasonal shedding of O157 results from introduction of new season forage crops. Most of the data to date have focused on adult dairy cattle and shown relatively minor impact on STEC shedding including O157. By comparing STEC shedding in adjacent pens with shared water troughs with shedding in adjacent pens lacking shared water troughs, the specific role of common water sources in STEC epidemiology was evaluated. The project is ongoing but the preliminary analysis, a general linear model of absolute STEC prevalence difference conditional on shared water trough or shared fence line, found no effect of shared water through on absolute prevalence difference. C. Salmonella Tennessee Evaluation of the use of probiotics and botanicals as feed additives to reduce Salmonella colonization in a chick model. Botanical treatments supported the growth of the probiotics and were biocidal in vitro but had limited effectiveness in vivo. Feed inclusion concentrations are being adjusted for optimization. Kansas A randomized trial to assess whether enrofloxacin metaphylaxis for bovine respiratory disease affects fecal shedding of Salmonella and Campylobacter in feedlot cattle. Preliminary results indicate that metaphylactic administration of enrofloxacin in feedlot cattle did not significantly affect fecal prevalence of Salmonella and Campylobacter. Additionally, metaphylactic administration of enrofloxacin did not impact naladixic acid or ciprofloxacin susceptibilities of Salmonella isolated from feeder cattle feces. Washington Transcriptional profiling of Salmonella Enteritidis strains using RNA-Seq identifies genes consistently highly expressed in biologically relevant microenvironments: A core of 23 genes are consistently highly expressed in three highly pathogenic S. Enteritidis strains, regardless of growth microenvironments representing the avian host. The role of these genes in bacterial physiology and virulence are being investigated for development of new vaccines or antimicrobial agents Overexpression of catalytically inactive dimethyl adenosine transferase (KsgA) unveils contribution of KsgA to Salmonella Enteritidis physiology and virulence: The results support KsgA as a potential candidate for drug and/or vaccine development. Identification and characterization of the immunomodulatory activity of CpG motifs of Salmonella: Cystine-guanine (CpG) motifs are DNA sequences (6 bp) that can have strong immune-modulatory activities. We identified 256 unique CpG motifs within the Salmonella pan-genome. We then synthesized 256 oligodinucleotides (ODNs) each containing a single CpG motif repeated 3 times. This study comprehensively characterized the immune-modulatory activities of all the possible CpG motifs found within Salmonella pan-genome and identified a number of immunostimulatory motifs. These newly identified CpG motifs could serve as targets for the development of ODNs that stimulate immune responses against Salmonella in mammals and chickens and also to improve the efficacy of currently used vaccines. D. Brachyspira Minnesota Association of Brachyspira antibiograms with species and source. In general, the in vitro MICs of Brachyspira species were low for tiamulin and valnemulin; and high for lincomycin and tylosin. In contrast to B. hyodysenteriae and “B. hampsonii”, several B. pilosicoli and B. murdochii isolates showed high tiamulin MICs. The odds of an isolate having a low MIC if originating from a sow site were significantly higher than those originating from a finisher site. Most isolates with high MIC profiles originated from three swine production systems. Analysis of virulence-associated genes in known and novel Brachyspira species to develop pathogen-specific diagnostic assays. Based on the most frequently detected virulence genes and usefulness of sequences to discriminate pathogenic Brachyspira species, the nox gene was determined to be a useful target for design of diagnostic PCR assays. Species-specific PCR assays for B. hyodysenteriae, “B.hampsonii”, and the Brachyspira genus were developed and evaluated. Further, the B. hyodysenteriae and “B. hampsonii” primer sets were evaluated for their utility as a duplex PCR. E. Streptococcus suis Kansas Streptococcus suis virulence. The SSU0587 gene encodes a protein of 1,491 amino acids that possesses beta-galactosidase activity. Deleting SSU0587 from clinical SS2 isolate JX081101 caused a loss of both beta-galactosidase activity and adherence to microvascular endothelial cells and reduced concentration in the brains of infected mice. F. Calicivirus Kansas Cathepsin L plays a crucial role in the replication of caliciviruses. This conclusion was supported by various evidence using recombinant cathepsin, diverse calicivirus, and cathepsin L inhibitors. Cathepsin L could be a therapeutic target as demonstrated in severe acute respiratory syndrome coronavirus and Ebola virus. Design, synthesis, and bioevaluation of viral 3C and 3C-like protease inhibitors. Among the lead compounds, a class of tripeptidyl transition state inhibitors were further characterized and validated. Two tripeptidyl compounds possess a potential as a broad range antiviral agents G. Rotavirus Ohio Probiotics: effects on neonatal innate immune responses, immune homeostasis, RV infections and vaccine efficacy. We demonstrated that in the neonatal Gn pig disease model, selected G+ and G-probiotics exert their beneficial effects by promoting immunomaturation and immune responses and regulating immune homeostasis, thereby modulating responses to virulent HRV and alleviating diarrhea. Our findings will contribute to alternative low cost probiotic treatments applicable to neonates (or mothers) to moderate RV disease, enhance oral vaccine efficacy, and reduce neonatal morbidity and mortality. Comparative in vivo and in vitro studies of porcine rotavirus G9P and human rotavirus Wa (G1P) in gnotobiotic pigs: Our comparative studies suggest that PRV G9P may replicate in immune swine cells which may facilitate its more efficient dissemination and persistence in swine. These findings contribute basic knowledge about RV replication and the virus associated immunopathology. Wyoming An improved method to capture and detect viruses from bioaerosols has been developed. H. Cryptosporidium parvum Illinois Our results suggest microneme secretion is a potential target that could be exploited for development of new drugs for the treatment of not only cryptosporidiosis but also other serious apicomplexan diseases such as toxoplasmosis. Objective 3. Focus on disseminating knowledge: We will provide training or continuing education to disseminate new information to students, producers, veterinarians, diagnostic labs and others to implement interventions and preventative measures. The PIs and Graduate Students involved in the project have been continuing to give presentations and updates enteric diseases and food safety at various scientific, veterinary, and diagnostic meetings in the previous year. They have effectively disseminated new information, reagents, and procedures to producers, industries, veterinary diagnostic laboratories and veterinarians. They also have generated many high impact peer-reviewed journal articles. The members have actively organized various outreach/education activities, such as 1) Dr. Armstrong organized the 5th annual University of Arizona Food Safety Conference on October 10, 2014; 2) Dr. Irene Hanning presented Webinar on Food Safety in Organic Poultry via eXtension Archived at http://www.extension.org/pages/25242 (National, 2014); 3) two half day symposia were organized by Dr. Linda Mansfield (MSU) to discuss scientific progress. Fall seminar series in Large Animal Clinical Sciences at the College of Veterinary Medicine at MSU where preharvest food safety was prominently featured. People attending came from the Agricultural, Veterinary Medicine, Microbiology and Food Science, and Human Nutrition departments. Objective 4. Group interaction: The group will interact in a variety of ways to facilitate progress including direct collaborations with joint publications, sharing of resources (pathogen strains, gene sequences, statistical analysis, bioinformatics information/expertise), and friendly feedback and facilitation for all research efforts at annual meetings. • NC-1202 members and their students presented their work in numerous national and international meetings. We held annual NC1202 meetings in Dec of 2014, and also sponsored 3 student awards for best oral and poster presentations at the Conference of Research Workers in Animal Disease meeting. • NC-1202 members have established active collaborations, partly reflected by joint grant/publications. Following are three active integrated food safety projects in which our NC1202 members are project directors and involve several NC1202 members at the participating institutions as co-project directors: 1) Zhang, Q. (PD, Iowa State Univ.) and other Co-PDs who are NC1202 members: Jun Lin (Univ. of Tennessee), Gireesh Rajashekara (Ohio State Univ). USDA NIFA Food Safety Challenge Grant, Novel Approaches for Mitigation of Campylobacter in Poultry. $2,500,000. Award period: 07/01/2012 – 06/30/2017. 2) Law, B. (PD, Univ. of Arizona) and other Co-PIs who are NC1202 members: Roy Curtiss III is a scientific advisor, and Ken Roland from the Curtiss lab is the Co-PI. USDA NIFA Food Safety Challenge Grant, The Development of an Efficacious Vaccine to Reduce Campylobacter in Chickens. $2,500,000. Award period: 8/1/2012 – 7/31/2017. 3) Moxley, R. (PD, Univ. of Nebraska) and T.G. Nagaraja and David Renter (Co-PDs, Kansas State Univ.) USDA NIFA Coordinated Agricultural Program Award, Shiga-toxigenic Escherichia coli (STEC) in the Beef Chain: Assessing and Mitigating the Risk by Translational Science, Education and Outreach. Total funding: $25,000,000 for 16 institutions, 51 collaborators. Award period: 1/1/2012 – 12/31/2016

Peer-reviewed journal articles • Zuowei Wu, Rachel Sippy, Orhan Sahin, Paul Plummer, Ana Vidal, Diane Newell, and Qijing Zhang. 2014. Genetic diversity and antimicrobial susceptibility of Campylobacter jejuni isolates associated with sheep abortion in the United States and the Great Britain. J Clin Micro, 52:1853-61. PMID: 24648552. • Zuowei Wu, Orhan Sahin, Fei Wang, Qijing Zhang. 2014. Proteomic identification of immunodominant membrane-related antigens in Campylobacter jejuni associated with sheep abortion. Journal of Proteomics, 99:111-122. PMID: 24487037. • Orhan Sahin, Eric R. Burrough1, Nada Pavlovic, Tim S. Frana, Darin M. Madson, and Qijing Zhang. 2014. Campylobacter jejuni as the cause of canine abortions in the Midwest United States. J Vet Diagn Invest, 26:699-704. PMID: 25085872. • Ma L, Wang Y, Shen J, Zhang Q, Wu C. 2014.Tracking Campylobacter contamination along a broiler chicken production chain from the farm level to retail in China. Int J Food Microbiol. 181:77-84. PMID: 24831929. • Wang Y, M. Zhang, F. Deng, Z. Shen, C. Wu, J. Zhang, Zhang Q, and J. Shen. 2014. Emergence of multidrug-resistant Campylobacter with a horizontally acquired ribosomal RNA methylase. Antimicrob. Agents Chemother. 58(9):5405-5412. • Ma, L., Z. Shen, G.W. Naren, H. Li, X. Xia, C. Wu, J. Shen, Q. Zhang, and Y. Wang. Identification of a novel G2073A mutation in 23S rRNA in amphenicol-selected Campylobacter jejuni. PLoS One. 9(4):e94503. doi: 10.1371/journal.pone.0094503 • Su, C.C., A. Radhakrishnan, N. Kumar, F. Long, J. R. Bolla, H.T. Lei, J. A. Delmar, S. V. Do, T.H. Chou, K. R. Rajashankar, Q. Zhang and E. W. Yu. 2014. Crystal structure of the Campylobacter jejuni CmeC outer membrane channel. Protein Sci. 23:954-961. • Shen, Z., T. Luangtongkum, Z. Qiang, B. Jeon, L. Wang, and Q. Zhang. 2014. Identification of a novel membrane transporter mediating resistance to organic arsenic in Campylobacter jejuni. Antimicrob Agents Chemother. 58(4):2021-2029. • Oh, E., Q. Zhang, and B. Jeon. 2014. Target optimization for peptide nucleic acid (PNA)-mediated antisense inhibition of the CmeABC multidrug efflux pump in Campylobacter jejuni. J. Antimicrob. Chemother. 69(2):375-380. • Qin, S.S., Y. Wang, Q. Zhang, M. Zhang, F. Deng, Z. Shen, C. Wu, S. Wang, J. Zhang, and J. Shen, 2014. Report of ribosomal RNA methylase gene erm(B) in multidrug resistant Campylobacter coli. J. Antimicrob. Chemother. 69 (4): 964-968. • Lin, J., K. Nishino, M.C. Roberts, M. Tolmasky, R.I. Aminov, and L. Zhang. 2014. Mechanisms of antibiotic resistance. Frontiers in Microbiology (section Antimicrobials, Resistance and Chemotherapy) Accepted • Nosanchuk, J.D., J. Lin, R. P. Hunter, R. I. Aminov. 2014. Low dose antibiotics: current status and outlook for the future. Frontiers in Microbiology (section Antimicrobials, Resistance and Chemotherapy). 5:478. doi; 10.3389/fmicb.2014.00478 • Lin, J. 2014. Antibiotic growth promoters enhance animal production by targeting bile salt hydrolase and its producers. Frontiers in Microbiology (section Antimicrobials, Resistance and Chemotherapy). 5:33. doi: 10.3389/fmicb.2014.00033 • Wu, J.E, J. Lin, and Q. Zhong. 2014. Physical and antimicrobial characteristics of thyme oil emulsified with soluble soybean polysaccharide. Food Hydrocolloids. 39:144-150. • Smith, K., X. Zeng, J. Lin. 2014. Discovery of bile salt hydrolase inhibitors using an efficient high-throughput screening system. PLOS One 9(1):e85344. DOI: 10.1371/journal.pone.0085344 • Zhang, H.W., X. Zeng, Q.Qi, K.L. Sun, C.J.Ma, X.J. Hu, J. Lin. 2014. The interaction of CmeA and CmeC is independent of substrate and CmeB in Campylobacter jejuni CmeABC system. Journal of Chinese Science Bulletin (In Press) • Zeng, X., S. Brown, B. Gillespie, J. Lin. 2014. A single nucleotide in promoter modulates the expression of the ?-lactamase OXA-61 in Campylobacter jejuni. Journal of Antimicrobial and Chemotherapy. 69:1215-1223. DOI:10.1093/jac/dkt515 • Andino, A., S. Pendleton, N. Zhang, W. Chen, F. Critzer, and I. Hanning. 2014. Survival and virulence of Salmonella spp. in poultry feed in strain and serovar dependent. Poult Sci. 93: 441-447 • Gonzalez-Gil, F., S. Diaz-Sanchez, S. Pendleton, A. Andino, N. Zhang, C. Yard, N. Crilly, F. Harte, and I. Hanning. 2014. Yerba Mate Enhances Probiotic Bacteria Growth In Vitro but as a Feed Additive does not Reduce Salmonella Enteritidis Colonization In Vivo. 93: 434-440. 47. • Diaz-Sanchez, S., D. D’Souza, D. Biswas, and I. Hanning. Botanical Alternatives to Antibiotics for use in Organic Poultry Production. Poultry Science. In Press • Salaheen, S., N. Chowdhury, I. Hanning, and D. Biswas. 2014. Zoonotic Bacterial Pathogens and Mixed Crop-Livestock Farming. Poultry Science. In Press • Kandasamy, S., K.S. Chattha, A.N. Vlasova, and L.J. Saif. 2014. Prenatal vitamin A deficiency impairs adaptive immune responses to pentavalent rotavirus vaccine (RotaTeq®) in gnotobiotic pig model. Vaccine 32:816-24. • Lee,C.S., C. Lee, J. Marion, Q. Wang, L.J. Saif and J. Lee. 2014. Occurrence of human enteric viruses at freshwater beaches during swimming season and its link to water inflow. Science of the Total Environment 472:757-766. • Jung, K, K. Scheuer, Z. Zhang, Q. Wang, L. J. Saif. 2014. Pathogenesis of GIII.2 bovine norovirus, CV186-OH/00/US strain in gnotobiotic calves. Vet Microbiol. 168:202-207. • Takanashi, S., L.J. Saif, J. H. Hughes, T. Meulia, K. Jung, K. A. Scheuer, and Q. Wang. 2014. Failure of propagation of human norovirus in intestinal epithelial cells with microvilli grown in three-dimensional cultures. Arch. Virol. 159:257-266. • Amimo, J.O, Okoth, E, Junga, J O, Ogara, W.O, Njahira, M.N, Wang, Q ,Vlasova, A.N, Saif, L.J, and Djikeng, A. 2014. Molecular detection and genetic characterization of Kobuviruses and Astroviruses in asymptomatic local pigs in East Africa. Arch Virol . • Gebreyes, W.A., J. Dupouy-Camet, M.J. Newport, C.J.B. Oliveira, L.S. Schlesinger, Y.M. Saif, S. Kariuki, L.J. Saif, W. Saville, T. Wittum, A. Hoet, L.J. King, S. Quessy, R. Kazwala, B. Tekla, T. Shryock, M. Bisesi, P. Pacthanee and S. Bonnmar. 2014. The global one health paradigm: Challenges and opportunities for tackling infectious diseases at the human, animal and environment interface in low-resource settings. PLOS Neglected Tropical Diseases (accepted). • Kandasamy, S., K.S. Chattha, A.N. Vlasova, G. Rajashekara, and L.J. Saif. 2014. Dual-colonization of Lactobacillus rhamnosus strain GG and Bifidobacterium animalis subsp. lactis Bb12 probiotics enhances mucosal B cell responses to an oral human rotavirus vaccine in a neonatal gnotobiotic pig model. Gut Microbes (in press). • Kumar, A., A N. Vlasova, Z Liu, K S. Chattha, S Kandasamy, M Esseili, X Zhang, G Rajashekara, and L.J. Saif. 2014. In vivo gut Transcriptome Responses to Lactobacillus rhamnosus GG and Lactobacillus acidophilus in Neonatal Gnotobiotic Piglets. Gut Microbes (submitted). • Hemida, M.G., R.A. Perera, P. Wang, M.A. Alhammadi, L.Y. Siu, M. Li, L.L. Poon,L.J. Saif, A. Alnaeem and M. Peiris. 2013. Middle East respiratory syndrome (MERS) coronavirus seroprevalence in domestic livestock in Saudi Arabia, 2010 to 2013. www.eurosurveillance.org. • Jung, K., Q. Wang, K.A. Scheuer, Z. Lu, Y. Zhang, and L.J. Saif. 2014. Pathology of US porcine epidemic diarrhea virus strain PC21A in gnotobiotic pigs. Emerg Infect Dis. http://dx.doi.org/10.3201/eid2004.131685. • Oka, T., L. J. Saif, D. Marthaler, M. A. Esseili, Meuli, T., C. M. Lin, A. N. Vlasova, K. Jung, Y. Zhang, and Q. Wang. 2014. Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene. Veterinary microbiology. DOI: 10.1016/j.vetmic.2014.08.012. (online at http://authors.elsevier.com/sd/article/S0378113514003885). • Martella, V., P. Pinto, E. Lorusso, B. Di Martino, Q. Wang, V. Larocca, A. Cavalli, M. Camero, N. Decaro, K. Banyai, L.J. Saif, and C. Buonavoglia. 2014. Detection and full-length genome characterization of novel canine vesiviruses. Emerging Infectious Diseases (submitted). • Kassem II, Rajashekara G. Formate Dehydrogenase Localization and Activity Are Dependent on an Intact Twin Arginine Translocation System (Tat) in Campylobacter jejuni 81-176. Foodborne Pathog Dis. 2014 Sep 30. [Epub ahead of print] PubMed PMID: 25268895. • Kashoma IP, Kumar A, Sanad YM, Gebreyes W, Kazwala RR, Garabed R, Rajashekara G. Phenotypic and Genotypic Diversity of Thermophilic Campylobacter spp. in Commercial Turkey Flocks: A Longitudinal Study. Foodborne Pathog Dis. 2014 Sep 3.[Epub ahead of print] PubMed PMID: 25184688. • Drozd M, Chandrashekhar K, Rajashekara G. Polyphosphate-mediated modulation of Campylobacter jejuni biofilm growth and stability. Virulence. 2014 Aug 15;5(6):680-90. doi: 10.4161/viru.34348. PubMed PMID: 25127528; PubMed Central PMCID: PMC4139409. • Malde A, Gangaiah D, Chandrashekhar K, Pina-Mimbela R, Torrelles JB, Rajashekara G. Functional characterization of exopolyphosphatase/guanosine pentaphosphate phosphohydrolase (PPX/GPPA) of Campylobacter jejuni. Virulence. 2014 May 15;5(4):521-33. doi: 10.4161/viru.28311. Epub 2014 Feb 25. PubMed PMID: 24569519; PubMed Central PMCID: PMC4063813. • Kassem II, Khatri M, Sanad YM, Wolboldt M, Saif YM, Olson JW, Rajashekara G. The impairment of methylmenaquinol:fumarate reductase affects hydrogen peroxidesusceptibility and accumulation in Campylobacter jejuni. Microbiologyopen. 2014 Apr;3(2):168-81. doi: 10.1002/mbo3.158. Epub 2014 Feb 7. PubMed PMID: 24515965; PubMed Central PMCID: PMC3996566. • Sanad YM, Jung K, Kashoma I, Zhang X, Kassem II, Saif YM, Rajashekara G. Insights into potential pathogenesis mechanisms associated with Campylobacter jejuni -induced abortion in ewes. BMC Vet Res. 2014 Nov 25;10(1):274. • Vogstad, A. R., R. A. Moxley, G. E. Erickson, T. J. Klopfenstein, and D. R. Smith. 2014. Stochastic simulation model comparing distributions of STEC O157 fecal shedding prevalence between cattle vaccinated with type III secreted vaccines and non-vaccinated cattle. Zoonoses Public Health 61:283-289. • Moxley, R. A., and G. R. Acuff. 2014. Peri- and post-harvest factors in the control of Shiga toxin-producing Escherichia coli in beef. Microbiol. Spectrum 2(6):EHEC-0017-2013. doi:10.1128/microbiolspec. EHEC-0017-2013. • Wijemanne, P., and R. A. Moxley. 2014. Glucose significantly enhances enterotoxigenic Escherichia coli adherence to intestinal epithelial cells through its effects on heat-labile enterotoxin production. PLoS One 9:e113230. • Wijemanne, P., J. Xing, E. M. Berberov, D. B. Marx, D. H. Francis, and R. A. Moxley. Relationship between heat-labile enterotoxin secretion capacity and virulence in wild type porcine-origin enterotoxigenic Escherichia coli strains. PLoS One (submitted) • Bisha B, Adkins JA, Jokerst JC, Chandler JC, Pérez-Méndez A, Coleman SM, Sbodio AO, Suslow TV, Danyluk MD, Henry CS, Goodridge LD. 2014. Colorimetric paper-based detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from large volumes of agricultural water. J Vis Exp. 9;(88). doi: 10.3791/51414. • Pérez-Méndez A, Chandler JC, Bisha B, Goodridge LD. 2014. Concentration of enteric viruses from tap water using an anion exchange resin-based method. J Virol Methods. 206:95-8. doi: 10.1016/j.jviromet.2014.05.025. • Pérez-Méndez A, Chandler JC, Bisha B, Goodridge LD. 2014. Evaluation of an anion exchange resin-based method for concentration of F-RNA coliphages (enteric virus indicators) from water samples. J Virol Methods. 204:109-15. doi: 10.1016/j.jviromet.2014.03.024. • Pérez-Méndez A, Chandler JC, Bisha B, Coleman SM, Zhanqiang S, Gang Y, Goodridge LD. 2013. Evaluation of a simple and cost effective filter paper-based shipping and storage medium for environmental sampling of F-RNA coliphages. J Virol Methods. 2013 Dec;194(1-2):60-6. doi: 10.1016/j.jviromet.2013.07.047. • Mirajkar N.S. and Gebhart C.J. Understanding the molecular epidemiology and global relationships of Brachyspira hyodysenteriae from swine herds in the United States: a multi-locus sequence typing approach. PLoS One. Sept 2014. 9(9):e107176. doi: 10.1371/journal.pone.0107176. • Vannucci FA, Gebhart CJ. Recent advances in understanding the pathogenesis of Lawsonia intracellularis infections. Vet Pathol. 2014 Mar;51(2):465-77. doi: 10.1177/0300985813520249. Epub 2014 Jan 29. PubMed PMID: 24476941. • Sampieri F, Allen AL, Alcorn J, Clark CR, Vannucci FA, Pusterla N, Mapes SM, Ball KR, Dowling PM, Thompson J, Bernstein LR, Gebhart CJ, Hamilton DL. Efficacy of gallium maltolate against Lawsonia intracellularis infection in a rabbit model. J Vet Pharmacol Ther. 2014 Dec;37(6):571-8. doi: 10.1111/jvp.12132. Epub 2014 Apr 15. PubMed PMID: 24730377. • Sampieri F, Alcorn J, Allen AL, Clark CR, Vannucci FA, Pusterla N, Mapes S, Ball KR, Dowling PM, Thompson J, Bernstein LR, Gebhart CJ, Hamilton DL. Pharmacokinetics of gallium maltolate in Lawsonia intracellularis-infected and uninfected rabbits. J Vet Pharmacol Ther. 2014 Oct;37(5):486-99. doi: 10.1111/jvp.12114. Epub 2014 Mar 15. PubMed PMID: 24628462. • Sampieri F, Vannucci FA, Allen AL, Pusterla N, Antonopoulos AJ, Ball KR, Thompson J, Dowling PM, Hamilton DL, Gebhart CJ. Species-specificity of equine and porcine Lawsonia intracellularis isolates in laboratory animals. Can J Vet Res. 2013 Oct;77(4):261-72. PubMed PMID: 24124268; PubMed Central PMCID: PMC3788657. • Sampieri F, Allen AL, Pusterla N, Vannucci FA, Antonopoulos AJ, Ball KR, Thompson J, Dowling PM, Hamilton DL, Gebhart CJ. The rabbit as an infection model for equine proliferative enteropathy. Can J Vet Res. 2013 Apr;77(2):110-9. PubMed PMID: 24082402; PubMed Central PMCID: PMC3605926. • Marthaler D, Jiang Y, Collins J, and Rossow K. Complete Genome Sequence of Strain SDCV/USA/Illinois121/2014, a Porcine Deltacoronavirus from the United States Genome Announc. March/April 2014 2:e00218-14; doi:10.1128/genomeA.00218-14 • Jindal N, Mor SK, Patnayak DP, Ziegler AF, Marthaler D, Goyal SM. Molecular characterization of turkey enteric reovirus S3 gene. Avian Pathol. 2014 Mar 25. PMID: 24666328 • Marthaler D, Rossow K, Culhane M, Goyal S, Collins J, Matthijnssens J, Nelson M, and Ciarlet M. Widespread Rotavirus H in Commercially Raised Pigs, United States. Emerg Infect Dis. 2014 Jul;20(7):1203-1206. doi: 10.3201/eid2007.140034. PubMed PMID: 24960190. • Marthaler D, Raymond L, Jiang Y, Collins J, Rossow K, and Rovira A. Rapid Detection, Complete Genome Sequencing, and Phylogenetic Analysis of Porcine Deltacoronavirus, United States. Emerg Infect Dis. 2014 Aug. http://dx.doi.org/10.3201/eid2008.14-0526. • Marthaler D, Suzuki T, Rossow K, Culhane M, Collins J, Goyal S, Tsunemitsu H, Kuga K, Ciarlet M, and Matthijnssens J. VP6 Genetic Diversity, Reassortment, Intragenic Recombination and Classification of Rotavirus B in Japanese and American Pigs. Vet Microbiol. 2014 Aug 27;172(3-4):359-66. doi: 10.1016/j.vetmic.2014.05.015. Epub 2014 Jun 2 • Alonso C, Goede DP, Morrison RB, Davies PR, Rovira A, Marthaler DG, Torremorell M. Evidence of infectivity of airborne porcine epidemic diarrhea virus and detection of airborne viral RNA at long distances from infected herds. Vet Res. 2014 Jul 14;45(1):73. • Vlasova A*, Marthaler D*, Wang Q, Culhane M, Rossow K, Rovira A, Collins J, and Saif L. Distinct Characteristics and Complex Evolution of PEDV Strains, North America, May 2013–February 2014. Emerg Infect Dis. 2014 Oct. http://dx.doi.org/10.3201/eid2010.140491 (*corresponding authors) • Marthaler D, Bruner L, Collins J, and Rossow K. Third Strain of Porcine Epidemic Diarrhea Virus, United States. Emerg Infect Dis. Emerg Infect Dis. 2014 Dec. http://dx.doi.org/10.3201/eid2012.140908 • Oka T, Saif L, Marthaler D, Esseili M, Meulia T, Lin C, Vlasova A, Jung K, Zhang Y, and Wang Q. Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene. Vet Microbiol. 2014 Aug 27. pii:S0378-1135(14)00388-5. doi: 10.1016/j.vetmic.2014.08.012. PubMed PMID: 25217400. • Marthaler D, Homwong N, Rossow K, Culhane M, Goyal S, Collins J, Matthijnssens J, and Ciarlet M. Rapid detection and high occurrence of porcine rotavirus A, B, and C by RT-qPCR in diagnostic samples. J Virol Methods. 2014 Sep 4;209C:30-34. doi: 10.1016/j.jviromet.2014.08.018. PubMed PMID: 25194889. • Li M, Monaco MH, Wang M, Comstock SS, Kuhlenschmidt TB, Fahey GC, Jr., et al. Human milk oligosaccharides shorten rotavirus-induced diarrhea and modulate piglet mucosal immunity and colonic microbiota. ISME J. 2014;8(8):1609-20. doi: 10.1038/ismej.2014.10. PubMed PMID: 24522264. • Tang X, Kuhlenschmidt TB, Li Q, Ali S, Lezmi S, Chen H, et al. A mechanically-induced colon cancer cell population shows increased metastatic potential. Molecular cancer. 2014;13:131. doi: 10.1186/1476-4598-13-131. PubMed PMID: 24884630; PubMed Central PMCID: PMC4072622 • Zhou M, Guo Z, Yang Y, Duan Q, Zhang Q, Yao F, Zhang X, Hardwidge PR, Zhu G, Flagellin and F4 fimbriae have Opposite Effects on Biofilm Formation and Quorum Sensing in F4ac+ Enterotoxigenic Escherichia coli, Veterinary Microbiology, 2014, Jan 10;168(1):148-53. PMID: 2423866 • Rüter C, Hardwidge PR. “Drugs from Bugs”: Bacterial effector proteins as promising biological (immune-) therapeutics, FEMS Microbiology Letters, 2014, 351:126-132. PMID: 24261744 • Feuerbacher LA, Hardwidge PR. Influence of NleH effector expression, host genetics, and inflammation on Citrobacter rodentium colonization in mice, Microbes and Infection, 2014, May;16(5):429-33. PMID: 2461320 • Zhou M, Guo Z, Duan Q, Hardwidge PR, Zhu G. Escherichia coli type three secretion system 2: a new kind of T3SS?, Veterinary Research, 2014, Mar 19;45(1):32. PMID: 24641581 • Tang Y, Zhang X, Yin Y, Hardwidge PR, Fang W. Streptococcus suis Type 2 SSU0587 protein is a beta-galactosidase that contributes to bacterial adhesion but not to virulence in mice, Journal of Veterinary Medical Science, 2014, 76(7): 1055–1059, 2014. PMID: 24670993 • Tang Y, Li F, Tan B, Liu G, Kong X, Hardwidge PR, Yin Y. Enterotoxigenic Escherichia coli infection induces intestinal epithelial cell autophagy, Veterinary Microbiology, 2014, Jun 25;171(1-2):160-4. PMID: 24742948 • Yang Y, Zhou M, Hou H, Zhu J, Yao F, Zhang X, Zhu X, Hardwidge PR, Zhu, G. Quorum sensing gene luxS regulates flagella expression and Shiga-like toxin production in F18 Escherichia coli, Canadian Journal of Microbiology, 2014, Jun;60(6):355-61. PMID: 24848979 • Tang Y, Tan B, Xiong X, Li F, Ren W, Kong X, Qui W, Hardwidge, PR, Yin Y. Methionine deficiency reduces autophagy and accelerates death in intestinal epithelial cells infected with enterotoxigenic Escherichia coli, Amino Acids, 2014, Epub ahead of print. PMID: 24965529 • Ren W, Liu G, Yin J, Chen S, Li T, Kong X, Peng Y, Yin Y, Hardwidge PR. Draft genome sequence of enterotoxigenic Escherichia coli strain W25K, Genome Announcements, 2014, Jun 26;2(3). PMID: 24970825 • Ren W, Yin J, Duan J, Liu G, Zhu X, Chen S, Li T, Wang S, Tang Y, Hardwidge PR. Mouse jejunum innate immune responses altered by enterotoxigenic Escherichia coli (ETEC) infection, Microbes and Infection, 2014, Epub ahead of print • Martinez-Quiles N, Feuerbacher LA, Benito-Leon M, Hardwidge PR. Contribution of Crk adaptor proteins to host cell and bacteria interactions, BioMed Research International, Advances on Host Cells-Bacteria Interaction by Enterobacteriaceae Pathogens, 2014, in press • Agga G, Scott HM, Amachawadi R, Nagaraja TG, Vinasco J, Bai J, Norby B, Renter DG, Dritz S, Nelssen J, Tokach M. Effects of chlortetracycline and copper supplementation on antimicrobial resistance of fecal Escherichia coli from weaned pigs. Prev Vet Med. 2014; 114(3–4): 231–246. • Cernicchiaro N, Renter DG, Cull CA, Paddock ZD, Shi X, Nagaraja TG. Fecal shedding of non-O157 serogroups of Shiga toxin-producing Escherichia coli in feedlot cattle vaccinated with an E. coli O157:H7 SRP® vaccine and fed a Lactobacillus-based direct-fed microbial J Food Prot. 2014; 5: 732-737. • Paddock ZD, Renter DG, Cull CA, Paddock ZD, Bai J, Nagaraja TG. Escherichia coli O26 in feedlot cattle: fecal prevalence, isolation, characterization and effects of an E. coli O157 vaccine and a direct-fed microbial. Foodborne Pathog Dis. 2014; 11(3): 186-193. • Smith R, Sanderson MW, Jones R, Renter D, Larson RL. Economic risk analysis model for Bovine Viral Diarrhea Virus biosecurity in cow-calf herds. Prev Vet Med. 2014; 113(4): 492-503. • Jacob M, Bai J, Renter DG, Rogers A, Shi X, Nagaraja TG. Comparing real-time and conventional PCR to culture-based methods for detecting and quantifying Escherichia coli O157 in cattle feces. J Food Prot. 2014; 77(2): 314-319. • Shivanna V, Kim Y, Chang KO. 2014. The crucial role of bile acids in the entry of porcine enteric calicivirus. Virology 456-457:268-278. • Shivanna V, Kim Y, Chang KO. 2014. Endosomal acidification and cathepsin L activity is required for calicivirus replication. Virology 464-465C:287-295. • Prior AM, Kim Y, Weerasekara S, Moroze M, Alliston KR, Uy RA, Groutas WC, Chang KO, Hua DH. 2013. Design, synthesis, and bioevaluation of viral 3C and 3C-like protease inhibitors. Bioorganic & Medicinal Chemistry Letters 23:6317-6320. • Samuelson DR, Eucker TP, Bell JA, Dybas LA, Mansfield LS, Konkel ME. 2013. The Campylobacter jejuni CiaD effector protein activates MAP 1 kinase signaling pathways and is required for the development of acute disease. Cell Communication and Signaling 2013, 11:79, doi:10.1186/1478-811X-11-79. • Malik A., Sharma D., St. Charles J.L., Dybas L.A., Mansfield L.S. 2013. Contrasting immune responses mediate Campylobacter jejuni induced colitis and autoimmunity, Nature Mucosal immunology, 13 November 2013; doi: 10.1038/mi.2013.97. • Jerome, JP and Mansfield LS. 2013 Within-host evolution of Campylobacter jejuni, In "Campylobacter Ecology and Evolution", Samuel K. Sheppard; Associate editor: Guillaume Méric, Caister Academic Press, UK, pp. 1-25, Publication date: April 2014, ISBN: 978-1-908230-36-2. • Flies AS, Maksimoski MT, Mansfield LS, Weldele ML and Holekamp KE. 2014. Characterization of toll-like receptors 1-10 in spotted hyenas. Veterinary Research Communications, Veterinary Research Communications Jun;38(2):165-70. doi: 10.1007/s11259-014-9592-3. Epub 2014 Feb 2. • Flies AS, Mansfield LS, Grant CK, Weldele ML, Holekamp KE. 2014. Markedly elevated antibody responses in wild versus captive spotted hyenas show that environmental and ecological factors are important modulators of immunity, Developmental and Comparative Immunology, in review. • Afema JA, Mather AE, Sischo WM. Antimicrobial Resistance Profiles and Diversity in Salmonella from Humans and Cattle, 2004-2011. Zoonoses Public Health. Nov 21. doi: 10.1111/zph.12172. [Epub ahead of print], 2014. • Miller CB, Pierle SA, Brayton KA, Ochoa JN, Shah DH, Lahmers KK. Transcriptional profiling of cross-protective Salmonella enterica serovar Typhimurium UK-1 dam mutant identifies a set of genes more transcriptionally active compared to wild-type, and stably transcribed across biologically relevant microenvironments. Pathogens. 3:417-436, 2014. • Addwebi TA, Call DR, Shah DH. Contribution of Salmonella Enteritidis virulence factors to intestinal colonization and systemic dissemination in day-old chickens. Poultry Science. 93:871-881, 2014. • Shah DH. RNA-seq reveals transcriptional differences between high- and low-pathogenic Salmonella Enteritidis strains. Applied and Environmental Microbiology. 80:896-906, 2014. • Paul NC, Al-Adwani S, Crespo R, Shah DH. Evaluation of passive immunotherapeutic efficacy of hyperimmunized egg yolk powder against intestinal colonization of Campylobacter jejuni in chickens. Poultry Science. 93:2779-87, 2014. • Eberhart LJ, Ochoa J, Besser TE, & Call DR. 2014. Microcin MccPDI reduces the prevalence of susceptible Escherichia coli in neonatal calves. Journal of Applied Microbiology 117:340-6. • Volkova VV, Lu Z, Besser TE, & Grohn Y. 2014. Modeling Infection Dynamics of Bacteriophages in Enteric Escherichia coli: Estimating the Contribution of Transduction to Antimicrobial Gene Spread. Applied and Environmental Microbiology 80:4350-62. • Suthar N, Roy S, Call DR, Besser TE, & Davis MA. 2014. An individual-based model of transmission of resistant bacteria in a veterinary teaching hospital. PLoS One 9(6)e98589. • Jaros P, Cookson AL, Campbell DM, Duncan GE, Prattley D, Carter P, Besser TE, Shringi S, Hathaway S, Marshall JC, French NP. (Accepted 2014). Geographic divergence of bovine and human Shiga toxin-producing Escherichia coli O157:H7 genotypes in New Zealand. Emerging Infectious Diseases. • Besser TE, Schmidt C, Shah DH, & Shringi S. (accepted 2014). Pre-harvest' Food Safety for E. coli O157 and other pathogenic STECs. Microbiology Spectrum. • Suthar N, Roy S, Call DR, Besser TE, Davis MA. An individual-based model of transmission of resistant bacteria in a veterinary teaching hospital. PLoS One. 2014 Jun 3;9(6):e98589. doi: 0.1371/journal.pone.0098589. eCollection 2014. PMID: 24893006 Book Chapters • Zeng, X., and J. Lin. 2014. Siderophore-mediated iron acquisition for Campylobacter infection. Chapter 10. Pp111- 124 In S. Sheppard and G. Meric (eds), Campylobacter Ecology and Evolution. Caister Academic Press, Norfolk, U.K. • Moxley, R. A., and G. R. Acuff. Peri- and post-harvest factors in the control of Shiga toxin-producing Escherichia coli in beef. Enterohemorrhagic Escherichia coli, 2nd Ed., American Society for Microbiology Press, Washington, D.C. (in press)