Brief Summary of Minutes of Annual Meeting

1. Qijing Zhang, Chair of NC-1202, gave an update on the renewed proposal. The proposal is accessible on the web site for NC-1202. Anybody interested in the proposal can print out a copy directly from the web page. 2. Administrative Advisor, Dr. Stromberg, congratulated everyone for the successful renewal of the group. The group was given a new NC number; NC1202. He made a presentation on overview of the expectation for the groups and submitting annual report and encouraged new memberships. 3. Appendix E uploading. Members of NC-1202 are required to submit the Appendix E form via the NIMSS system. Members dont have to be state representatives. Investigators from medical schools are allowed and encouraged to become member of NC-1202. 4. Gireesh Rajashekara was elected as the secretary and will become the Chair of NC1202, replacing Lynn Joens on the organizing committee. 5. New members were introduced. 6. Student awards. NC-1202 offers awards for students to compete in the Gastroenteric Disease Section of CRWAD. This year two awards will be offered, one for oral presentation and another for poster presentation. Radhey Kaushik serves as Chair for the student award selection committee. He will continue to serve as Chair for the next year. Selection of a co-chair working with him was accomplished. Historically, the student awards were funded by annual registration fee. How to increase the number of abstracts submitted to the Gastroenteric Disease Section was discussed. One suggestion was to offer student travel awards or increase the dollar amount of the awards. This requires industry sponsors and contact with industries for possible support of NC-1202 activities was proposed. There was also a discussion on changing the registration fee to annual dues, which will ensure a steady source of income for the committee. 7. There was discussion on naming the student awards. The group unanimously agreed to name the awards for Lynn Jeons and David Francis, respectively, in honor of their scientific accomplishments and their dedication to NC1202. 8. Proposed title change for the Gastroenteric Disease Section of CRWAD. There was a motion to change the section name from Gastroenteric Disease to a new name. Dr. Rod Moxley will discuss with CRWAD executive committee to finalize the name for the section in the future CRWAD meetings. The rationale for the change is that many members of NC-1202 work with zoonosis and food safety and this change will attract more submission of abstracts to the section. The attendee unanimously voted for the title change. Rodney Moxley will take this issue to the CRWAD Council for approval. 9. Annual report. Annual state reports are due to Qijing Zhang and those who have not submitted are asked to do so. Bert Stromberg emphasized the importance of outcomes and impact. These should be clearly stated in the report. 10. To reduce registration fee, the committee voted in favor of eliminating food items for future meetings.

Objective 1. Focus on emerging issue- 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. Campylobacter jejuni Michigan The LOS of C. jejuni can mimic gangliosides enriched on peripheral nerves leading to autoimmunity. Because up to 37.7% of dairy cattle have been found to shed C. jejuni, we sought to determine whether the calves were the source of the family infections. Preliminary typing results suggest that some calf and human C. jejuni isolates have similar characteristics. Draft Genome Sequences of 2 C. jejuni Clinical Isolates, NW and 2600. NW was isolated from a patient returning from Kenya with acute gastroenteritis. Strain D2600 was also isolated from a patient with gastroenteritis. These clinical isolates colonized C57BL/6 interleukin 10 knockout (IL-10-/-) mice without inducing a robust inflammatory response. Genome sequences of NW and D2600 will facilitate comparisons with strains that induce gastrointestinal inflammation in this mouse model. Iowa We compared the clinical isolates of C. jejuni clone (clone SA) from sheep abortions with the human isolates using PFGE, MLST, and serotyping. The results provide strong molecular and epidemiological evidence for zoonotic transmission of this emergent clone from ruminants to humans. Additionally, C. jejuni clone SA was identified in raw milk, cattle feces, feces and bile of healthy sheep, and abortion cases of cattle and goats, indicating the broad distribution of this pathogenic clone in ruminants. Comparative genomics analysis of C. jejuni isolates associated with sheep abortion in the U.S. and the U.K. and non-abortifacient isolates have been performed. Our findings indicated the presence of several genes/gene clusters that were uniquely/predominantly associated with the clone SA isolates. Using one of these genes, we have developed a specific and sensitive rapid screening/identification technique for clone SA (loop-mediated isothermal amplification method). Additionally adaptive changes in transcriptome of clone SA upon exposure to host specific factors are being determined. Ohio Here, we determined the prevalence, genotypic, and phenotypic properties of Campylobacter isolated from fecal samples of dairy cattle and starlings our results highlight starlings as potential reservoirs for C. jejuni and may play important role in the epidemiology of clinically important C. jejuni in dairy population. B. Shiga toxin-producing E. coli (STEC) Kansas Our objective was to determine the prevalence of seven STEC O groups (O26, O45, O103, O111, O121, O145, and O157) and four major STEC virulence genes (stx1, stx2, eae and ehxA) in cohorts of commercial feeder cattle using selective culture methods, four types of IMS beads (O26, O103, O111 and O157), and an 11-gene multiplex PCR (mPCR). Within-cohort prevalence significantly (P < 0.05) varied for certain O-specific serogroup genes. Because cattle are known reservoirs of STEC, we investigated fecal carriage of E. coli O104 in US cattle. Our results suggest that E. coli O104 is present in cattle feces, but the strains do not appear to carry genes characteristic of the virulent hybrid strain. Fresh steam-flaked corn (SFC) is a hotspot for aggregation of house flies (HF) that likely deposit /acquire fecal bacteria, including E. coli O157, on/from this substrate. SFC was screened by a culturing approach for fecal coliforms before and after exposure to HF under the natural conditions in a cattle feedlot in Kansas. Our data show that HF likely play a role in contamination of SFC with E. coli and other enteric bacteria. Washington State We have developed a 48-plex chromosomal backbone SNP allele assay to efficiently type E. coli O157 isolates into the eight lineages as well as 4 sub-lineages. These SNP genotypes are strongly associated with the Stx-encoding bacteriophage chromosomal insertion site genotypes that we have used previously identify and differentiate clinical and bovine biased genotypes of this pathogen. Using the SNP assay, we have genotyped approximately 800 isolates of known sources, with the goal of evaluating seasons, production systems, hosts of origin, interventions, etc., for effects on clinical and bovine-biased genotypes. The most pronounced effect seen is that a high proportion (>90%) of isolates from feedlots belong to a single, clinical genotype that alone accounts for >2/3 of human infection. We are currently evaluating whether this genotype is favoured by feedlot (high grain) rations. We have investigated seasonal variation of O157 shedding by cattle using experimental inoculations to determine whether the variation seen is due to cattle factors, gastrointestinal factors, or extrinsic factors. Results indicate no significant seasonal variation in the amount or duration of shedding, consistent with seasonal variation being due to factors extrinsic to cattle. We are testing the hypothesis that high summertime seasonal shedding of O157 results from introduction of new season forage crops, by sampling pens of cattle just prior to and just after introduction of the new forage crops. The initial trials have showed consistent and sometimes dramatic increases in O157 shedding following these introductions. C. Caliciviruses Ohio We compared Porcine Sapovirus SaV resistance to physicochemical treatments to that of HuNoVs (by reverse transcription-PCR), FCV and MNV (by infectivity assays). The higher stability of SaV than FCV to heat and acid, and its higher resistance to UV light than FCV and MNV make this enteropathogenic virus a promising surrogate for HuNoVs in both in vivo and in vitro studies. We found that NoV VLPs bind to lettuce using multiple carbohydrates, suggesting that simple water washes may be insufficient to remove NoV contamination. We found that SaV RNA persisted with higher titers on outer leaves that were damaged vs. control non-damaged leaves for at least 21 days. However, the virus was not transferred from outer to inner leaves. We screened 1,874 fecal samples from clinically healthy finisher pigs in nine North Carolina swine farms during 2009 by RT-PCR-coupled hybridization assays using primers and probes specific for porcine NoVs or using RT-PCR and calicivirus universal primers. The overall prevalence of porcine NoVs was 18.9%, and we detected GIX SaVs for the first time in the US. We also determined the full-length genome sequence of porcine GII.18 NoV. Our study provides the first GII.18 strain full-genome sequence data which can be used for genomic comparisons and primer design and molecular manipulation (infectious clone) for future pathogenesis studies. D. Turkey and Chicken Enteric viruses Southeast Poultry Research Laboratory, ARS, USDA Using metagenomic approaches we identified a novel parvovirus from enteric content of chickens and turkeys that were affected by enteric diseases. Comparative sequence analysis showed that the chicken parvovirus (ChPV) and turkey parvovirus (TuPV) represented a new member in the Parvovirus family. Following experimental infection, two-day-old broiler chickens showed characteristic signs of enteric disease. Our data indicate that ChPV alone induces RSS in broilers and an important determinant in the complex etiology of enteric diseases of poultry. A comparative metagenome comparing the RNA viral metagenomes from a healthy flock and a sister flock affected by enteric disease was further analyzed. This analysis revealed numerous picornaviruses that were determined to be homologous to the avian turdiviruses and the turkey and duck hepatitis viruses. Further, molecular diagnostic assays targeting the picornavirus capsid protein gene were designed and tested using archived and field enteric samples. E. Rotavirus Ohio Fecal samples collected in different seasons in 2004 and 2011 from diarrheic and healthy nursing piglets from 5 selected swine farms in the US were screened for group (Gp) A, B and C rotavirus using RT-PCR. Our results indicate high genetic heterogeneity in RVC genes and the concurrent co-circulation of different genotypes. Our preliminary data indicate that there are multiple genotypes of RVA (not only G4 and G5 as in current vaccine) currently circulating in US swine. In conclusion, our study demonstrates that infection with distinct RVC is more frequent among nursing piglets than RVA and RVB infections. These findings are of utmost importance to design updated RV vaccines. F. Antimicrobial resistance Washington State The CTX-M families of resistance gene determinants have spread globally in human hospitals, mainly plasmid-borne in E coli and Klebsiella infections. Recently their incidence in companion animals and livestock has reportedly increased. We have detected a high prevalence of CTX-M positive fecal E. coli in dairy calves in Washington State and have determined that their emergence in Washington dairy animals probably occurred between 2008 and 2011. We are currently studying risk factors for their presence on 30 Washington dairy farms. An individual based model was developed to simulate spread of antibiotic resistant pathogens in a veterinary teaching hospital. Preliminary simulations indicated that the probability of transmission depends on the nature of each hospital site, patient length of stay and the frequency of decontamination. Tennessee Bacterial pathogens have co-evolved with host to counteract the effect of endogenous AMPs. However, molecular mechanisms of AMP resistance in Campylobacter are still largely unknown. Our study identified four genetic loci in Campylobacter that will be useful for characterizing molecular basis of Campylobacter resistance to AMPs, a significant knowledge gap in Campylobacter pathogenesis. Despite prevalent beta-lactam resistance in C. jejuni, the molecular basis of beta-lactam resistance in C. jejuni is still largely unknown. We performed an in vivo random transposon mutagenesis to identify genes required for beta-lactam resistance in C. jejuni 81-176. We identified a novel mechanism of beta-lactam resistance in C. jejuni, which will help us better understand the development and regulation of ²-lactam resistance, a significant issue in many bacterial pathogens. We hypothesize that gut microbiome of food animals also contain diverse and novel AR genes which may contribute to the emergence of AR in foodborne human pathogens. Our study showed the effectiveness of functional cloning approach for examination of AR reservoir in food animals, revealed novel AR resistance genes in gut microbiome of chickens from different production systems, and demonstrated functional compatibility of the novel AR genes in enteric pathogens. Iowa We identified a predicted arsenic efflux protein (ArsB) involved in inorganic arsenic resistance and a novel membrane transporter (named ArsP) that contributes to organic arsenic resistance in Campylobacter. The arsenic resistance profiles in C. jejuni isolates from different animal hosts including human, sheep, chicken, and turkey were found to be significantly different with all tested arsenic compounds. Specifically, the sheep isolates had the lowest level of arsenic resistance, while the isolates from turkey had the highest level of arsenic resistance. In another study, genetic mechanisms for differential expression of the CmeABC Efflux Pump in naturally occurring isolates of C. jejuni have been determined. Isolates from turkeys, broilers and humans were analyzed for CmeA, CmeB, and CmeC expression by immunoblotting and cmeB expression by real time RT-PCR. The adaptive mechanisms employed by C. jejuni to erythromycin treatment were studied. The transcriptomes of C. jejuni NCTC 11168 was determined after 30 min treatment with lethal or sub-lethal doses of Ery by competitive microarray hybridizations. Following the treatment with Ery, 139 genes were up-regulated and 119 were down-regulated. Interestingly, several gene operons with putative involvements in drug efflux were up-regulated after exposure to the lethal dose of Ery. Detailed functional analyses of these operons indicated that outer membrane/periplasmic proteins have a role in both oxygen tolerance and chicken colonization in C. jejuni. G. Lawsonia intracellularis Minnesota The aim of this study was to describe an alternative protocol for cultivation of L. intracellularis in cell monolayers. McCoy cell monolayers were heavily infected by L. intracellularis demonstrating the success of the alternative bag incubation protocol. Incubation in the bag provided environmental conditions for L. intracellularis to infect the cells, similar to the conventional protocol. Based on these preliminary results, we believe this approach can be used for static cultivation of bacteria without requiring a Tri-gas incubator. Objective 2. Focus on effective intervention- develop and improve interventions and preventative measures to reduce the incidence and prevalence of infections of cattle and swine with enteric and foodborne disease agents. A. C. jejuni Michigan We hypothesized that in the absence of the regulatory cytokine IL-10, uncontrolled induction of proinflammatory cytokines and infiltration of inflammatory cells is responsible for tissue obliteration in colitis. The work shows for the first time the adaptive immune response (Type 1 and 17 response) that leads to inflammatory bowel disease in this genetically susceptible mouse model. These results can be used to understand this process in humans. Future work is directed towards understanding the relative importance of Type 1 and 17 pathways and innate and adaptive lymphocytes in C. jejuni induced colitis. Lack of good, tractable small animal models has limited understanding of GBS pathogenesis and new therapies. Our results showed NOD, NOD B7-2-/-, and NOD IL-10-/- mice develop neurological signs including decreased activity, failure to rear, splayed feet, shortened stride length, foot drag, and hind limb paralysis among other signs. Sciatic nerve lesions were consistent with acute inflammatory demyelinating polyradiculoneuropathy seen in GBS. Ohio Whole genome expression studies showed that respiratory proteins (RPs) were differentially expressed under varying conditions and stresses, suggesting further unidentified roles for RPs in C. jejunis adaptation. Our studies suggest that the RPs may facilitate the prevalence of C. jejuni in a variety of niches, contributing to the pathogens remarkable potential for adaptation. The methylmenaquinol:fumarate reductase, Mfr of C.jejuni is a Tat-dependent periplasmic redox protein that contributes to the metabolism of fumarate. Since chemically oxidized redox-enzymes, contribute to the generation of oxidative stress in E. coli, we assessed the role of Mfr in C. jejunis resistance to hydrogen peroxide (H2O2). Our results suggested that Mfr contributes to H2O2 resistance in C. jejuni. The role of exopolyphosphatases (PPX/GPPA) in poly-P homeostasis and C. jejuni pathobiology remains unexplored. Here we characterized for the first time the role of PPX/GPPA proteins of C. jejuni. Our results expand our knowledge about the multifactorial regulation of poly-P and ppGpp metabolism, two critical molecules that modulate environmental stress responses and virulence in C. jejuni. Here we investigated the effect of nanoparticle encapsulated outer membrane proteins (OMPs) on C. jejuni colonization in chickens. Chicken were vaccinated nanoparticle encapsulated OMPs by different routes (subcutaneous (s. c) or oral) and doses of candidate vaccine. In conclusion, the s.c route of vaccination with nanoparticle encapsulated OMP of C. jejuni may serve as a candidate vaccine in control of C. jejuni colonization. Arizona We are continuing with our efforts to develop an efficacious vaccine using an attenuated Salmonella vector to express C. jejuni proteins shown to be involved in the colonization of broilers. The Salmonella expressing the CjLAJ3 protein was evaluated as vaccine. There was not a significant reduction of C. jejuni in cecal contents of chicks vaccinated with the vector expressed CjLAJ3 protein following challenge. We plan conduct two and three component vaccine studies to determine if a combination of multiple expressed proteins will further reduce the Campylobacter load in broilers. Tennessee We have characterized two Ferric enterobactin (FeEnt) receptors (CfrA and CfrB) in C. jejuni and C. coli, the enteric human pathogens that do not produce any siderophores. Together, our study identified and characterized a novel periplasmic trilactone esterase and suggested a new model of FeEnt acquisition. We also established that Campylobacter could utilize high-affinity salmochelin for iron acquisition, and provided insights into the delicate interaction between Campylobacter and host during infection. The USDA licensed live attenuated S. Typhimurium vaccine strain Ç8914 and vector pYA3493 were used for construction of recombinant Salmonella vaccine expressing CmeC or CfrA. In conclusion, we have successfully developed CfrA- and CmeC-based subunit vaccines, which provides us a solid foundation to evaluate different vaccination regimens for effective mitigation of Campylobacter in poultry in the future. The growth promoting effect of Antibiotic Growth Promoters has been shown to be highly correlated with the decreased activity of intestinal bile salt hydrolase (BSH), an enzyme that is produced by various gut microflora and involved in host lipid metabolism. Our study identified and characterized a BSH with broad substrate specificity from a chicken L. salivarius strain and established a solid platform for us to discover novel BSH inhibitors, the promising feed additives to replace AGPs for enhancing the productivity and sustainability of food animals. Washington State We have evaluated chicken egg-yolk derived antibodies against several C. jejuni chicken colonization associated proteins has opened opportunities for their use as novel passive immunotherapeutic agents to reduce C. jejuni loads in broiler chickens. B. Enterotoxigenic E. coli (ETEC) South Dakota In this study, we applied a genetic fusion strategy to construct a single antigen consisting of representative antigenic peptides from K88, F18, and LT, examined the fusion antigen vaccine development against ETEC associated diarrhea. Immunized piglets were protected when challenged with virulent ETEC strain. Results indicated that live E. coli strain expressing the K88-F18-LTA2:B fusion antigen can be developed as a broadly protective vaccine against porcine post-weaning diarrhea. We described a reproducible gnotobiotic piglet model of post-weaning ETEC diarrhea and efficacy tests of subunit vaccines consisting of K88 (F4) fimbriae and/or heat labile enterotoxin (LT) delivered by the intranasal route. The current study provides a system whereby various ETEC antigens and/or combinations of antigens can be tested in exploring strategies for the development of vaccines for ETEC. To test the ability of non-pathogenic E. coli constructs to exclude virulent ETEC sufficiently to prevent clinical disease, we utilized a piglet ETEC challenge model. This study shows for the first time that pre-inoculation with an avirulent strain expressing adhesive fimbriae and a non-toxic form of LT provides significant short term protection from challenge with a virulent ETEC strain that expresses the same fimbrial adhesion and enterotoxin. Significance of EAST1 in ETEC-associated diarrheal has not been determined, even though EAST1 is highly prevalent among ETEC strains. Results from this study indicated that EAST1 alone is not sufficient to cause diarrhea in five-day old gnotobiotic pigs, and suggest that EAST1 likely is not a virulence determinant in ETEC-associated diarrhea. Flagella of F18+ ETEC strains play a role in adherence and infection. The results from this study clearly demonstrate that flagella are required for efficient F18ab E. coli adhesion, invasion, biofilm formation, and IL-8 production in vitro Nebraska Here we determined whether ETEC exposure to glucose will affect its ability to adhere to intestinal epithelial cells via its effects on LT production. This is the first study demonstrating that glucose (at a specific concentration) in the culture medium, through its effects on LT production, significantly affects bacterial adherence, lending support to the hypothesis that the composition of dietary nutrients in the intestine of the host may directly influence the severity of ETEC infection. Kansas We investigated intestinal epithelial cell signal transduction pathways activated by ETEC and quantified the relative importance of these host pathways to enterotoxin (LT) -induced ETEC adherence. We show that ETEC activates both NF-kB and mitogen-activated protein kinase signalling pathways through mechanisms that are primarily dependent upon LT. Our findings provide insight into previously unexplored functions of LT and their relative importance to ETEC virulence. The extent to which ETEC subverts innate immune responses by directly targeting the NF-kB pathway is an understudied topic. We found that ETEC secretes a heat-stable, proteinaceous factor that blocks NF-kB signaling normally induced by TNF, interleukin-1â, and flagellin. These data suggest that ETEC evades the host innate immune response by directly modulating NF-kB signaling. C. Cryptosporidium parvum Illinois We hypothesized that L-PUFFA block a conserved apicomplexan microneme secretion pathway that regulates parasite motility. 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. To probe this hypothesis I have leveraged NC1041 funding by partnering with Dr. David Sibley, Washington University School of Medicine on an NIH grant entitled, Designing Selective Inhibitors of Calcium-Dependent Kinases in Parasites. D. Shiga toxin producing E. coli (STEC) Nebraska Mucosal explants from the colons of three slaughtered steerswere inoculated with 13 representative strains divided among STEC serogroups O26, O45, O103, O104, O111, O121, and O145. Adherence studies with the same strains were also done with human Caco-2 cells. To our knowledge, this is the first report of this organism being able to colonize bovine colonic epithelium. Also, based on the work reported herein, some STEC also have virulence mechanisms in place allowing them to invade colonic epithelial cells. Kansas Our objective was to determine whether commercially available pre-harvest interventions that were implemented to control fecal shedding of E. coli O157 in commercial feedlot cattle had an effect on fecal shedding of non-O157 STEC. By studying cattle that received one, both, or neither of two interventions  a siderophore receptor and porin proteins-based vaccine (VAC) and a direct-fed microbial product (DFM)  we found no significant treatment effects on fecal shedding of non-O157 STEC serogroups. Our objective was to determine whether the addition of starch to a dried distillers grains DDG-supplemented diet negates the effects of DG on fecal shedding of E. coli O157:H7. Our study confirms the positive association between feeding DG and fecal shedding of E. coli O157:H7 and starch addition to the DDG diet did not have effect on fecal shedding of E. coli O157:H7. We found that phosphorylation of RPS3 Ser209 by the kinase IKKb was crucial for nuclear localization of RPS3 in response to activating stimuli. Moreover, virulence protein NleH1 of E. coli O157:H7 specifically inhibited phosphorylation of RPS3 Ser209 and blocked RPS3 function, thereby promoting bacterial colonization and diarrhea but resulting in less mortality in a gnotobiotic piglet-infection model. Functional differences and interactions between the E. coli type III secretion system effectors NleH1 and NleH2. We found functional differences between NleH1 and NleH2 in their ability to regulate the host NF-kB pathway. Our data show that NleH1 and NleH2 have pronounced functional differences in their ability to alter host transcriptional responses to bacterial infection. E. Norovirus Ohio We tested ways to increase HuNoV infectivity in Gn pigs to create a more robust animal model for HuNoV. We also investigated the impact of an innate immunity mediator, IFN-± as an antiviral for oral treatment of HuNoV-infected Gn pigs. Simvastatin induced significantly earlier onset and longer duration of fecal virus shedding in treated pigs, frequently with higher fecal viral titers. Finally, in contrast to the increased HuNoV shedding that simvastatin induced, curtailed viral shedding was observed in HuNoV-infected pigs pre-treated with recombinant human IFN-± (rhIFN-±). This finding indicates that IFN-± has potential as an antiviral against HuNoV. Kansas The conserved key sites of 3Cpro or 3CLpro in Picorna, Noro, and Corona viruses (Supercluster) may serve as attractive targets for the design of broad-spectrum antivirals. We reported the structure-based design and synthesis of potent protease inhibitors of Norwalk virus (NV), a member of the Caliciviridae family. Furthermore we reported the broad-spectrum antiviral activities of three compounds possessing a common dipeptidyl residue with different warheads, i.e., aldehyde (GC373), a bisulfite adduct (GC376), and a ±-ketoamide (GC375), against viruses that belong to this supercluster. F. L. intracellularis Minnesota Non-pathogenic L. intracellularis variants have been obtained through multiple passages in cell culture but there is no information regarding the number of passages necessary to attenuate a pathogenic isolate. The present study evaluated the susceptibility of pigs to L. intracellularis after 10, 20 and 40 passages in vitro. These results demonstrate attenuation of the virulence properties of L. intracellularis between 20 and 40 cell passages in vitro. The goal of this study was to determine if feces from rabbits experimentally infected with L. intracellularis could be the source of infection for naïve weanling foals. The results support the role of rabbits as asymptomatic amplifier of L. intracellularis and serve as source of infection for susceptible foals. The objective of this study was to determine the efficacy of an avirulent L. intracellularis vaccine in preventing proliferative enteropathy in weanling foals. Intra-rectal administration of a commercial avirulentporcine vaccine against L intracellularis resulted in complete protection against proliferative enteropathy in the foals in this study This pilot study aimed to demonstrate the susceptibility of rabbits to L. intracellularis obtained from a clinical equine proliferative enteropathy case, and thus provide a potential animal model for investigations on pathogenesis and therapy in horses. This initial study for the development of an EPE in a rabbit model simulates natural infection, as typical lesions, immune response, and fecal shedding were present. F. Rotavirus Kansas In this study, we examined the roles of the key enzymes for cellular triglyceride TG synthesis (lipogenesis) in the replication of rotaviruses by using inhibitors of fatty acid synthase, long chain fatty acid acyl-CoA synthetase (ACSL). Triacsin C, a natural ACSL inhibitor from Streptomyces aureofaciens, was found to be highly effective against rotavirus replication. Many novel triacsin C analogs significantly reduced rotavirus replication. Our results suggest triacsin C and/or its analogs as potential therapeutic options for rotavirus infections. Ohio We investigated the impact of lactic acid bacteria colonization (Lactobacillus rhamnosus strain GG and Bifidobacterium lactis, probiotics) with/without mothers milk/cols on B cell antibody responses to an attenuated oral rotavirus (RV) vaccine in a gnotobiotic (Gn) pig model. Milk/cols did not affect probiotic colonization in RV vaccinated pigs. However milk/cols fed unvaccinated pigs shed higher probiotic bacterial titers than unvaccinated controls. In RV vaccinated pigs, milk/cols and probiotics enhanced mean serum IgA RV antibody titers and intestinal IgA antibody secreting cell (ASC) numbers compared to milk fed, non-probiotic supplemented pigs. We examined how prenatally acquired vitamin A deficiency (VAD) modulates innate and antibody responses and rotavirus (RV) vaccine efficacy in Gn pigs and challenged with virulent RV (VirRV) with or without vitamin A supplementation. Our results suggest that prenatal VAD causes an imbalance in innate immune responses and exacerbates VirRV infection. Vitamin A supplementation failed to alleviate the VAD-associated imbalance in innate immune responses or the exacerbated VirRV infection. Additionally, our results suggest that vitamin A metabolism may be affected in VAD piglets, leading to altered immune function. Illinois A sialyllactose-containing neoglycolipid was synthesized and assayed for its ability to inhibit rotavirus binding and infectivity of host cells. Sialyllactose was coupled to dipalmitoyl- phosphatidylethanolamine by reductive amination. Results of in vivo animal challenge studies using newborn pigs, demonstrated SLPE afforded complete protection from rotavirus disease. In collaboration with Dr. Sharon Donovan, we also have demonstrated selected natural human milk oligosaccharides, the third most abundant component in human milk, reduce the duration of diarrhea in piglets, possibly in part by promoting immunoglobulin response to rotavirus infection and modulating the gut microbiota. South East Poultry, USDA, ARS, Georgia In order to validate an institutional effluent decontamination system (EDS) in use at our facility, we determined the time and temperature necessary for the complete inactivation of common poultry enteric viruses. This investigation determined the thermal inactivation curve for rotavirus, reovirus and astrovirus in a buffered solution, as well as the endpoint thermal inactivation of these non-enveloped viruses in the EDS, which decontaminates effluent prior to disposal in the municipal sewer. This study confirmed the full inactivation of these viruses by the EDS. G. Salmonella Washington State We generated and screened >4000 mutants of a highly pathogenic S. Enteritidis strain in vitro for their virulence characteristics and identified several novel virulence associated genes. In addition, we are also evaluating the potential usefulness of the virulence-attenuated mutants as a live attenuated vaccine. We characterized more than 100 S. Enteritidis strains for their virulence characteristics and identified several strains that differ in their virulence and survival within egg albumen. We have performed global comparative transcriptome analysis using a next generation sequencing on multiple high and low-pathogenic strains of S. Enteritidis. This led to identification of several previously uncharacterized genes whose expression is upregulated in high-pathogenic strains. We screened a library of >20,000 mutants of a highly pathogenic strain of S. Enteritidis using oral infection in one-day old chickens as a model system and identified several genes that play a role in intestinal invasion and/or systemic dissemination of S. Enteritidis in chickens. Objective 3. Focus on disseminating knowledge Provide training or continuing education opportunities and dissemination of information to students, producers, veterinarians, and diagnostic laboratories Arizona Dr. Joens organized the University of Arizona Food Safety Conference on October 12, 2012, held at the Omni Tucson National Resort. Approximate attendance was 108 attendees, with 20% of attendees coming from food safety industries or government agencies. The Food Safety Conference consisted of 10 hours of contact time over one day, with seven invited speakers followed by a poster session and a dinner to facilitate dialogue and collaborative work. Minnesota The Principle Investigators and Students involved in the project have given presentations and updates on both swine and equine proliferative enteropathy at various scientific, veterinary, and diagnostic meetings in the previous year. These include the Leman Swine Conference, the Rushmore Symposium, the American Association of Equine Practitioners, the American College of Veterinary Internal Medicine and the American Association of Swine Veterinarians. Kansas Through several venues we have provided educational opportunities to professional and graduate students as well as continuing education to veterinarians on enteric pathogens in livestock production systems. The majority of the shared information has focused on the impacts of Shiga toxin-producing Escherichia coli and Salmonella in the beef industry, the conclusions that can be reached based on recent research, and the potential opportunities to reduce these pathogens in beef production systems. Michigan Dr. Mansfield organized a seminar in Food and Waterborne Diseases for the faculty and students of Michigan State University on October 19, 2012. People attending came from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science, and Human Nutrition departments. Dr. Mansfield helped to organize and attended the USDA Enteric Diseases Meeting NC1041 in Chicago, Illinois on December 1st and 2nd. Dr. Mansfield gave an invited talk entitled Murine models of the autoimmune neuropathy Guillain Barré Syndrome for the Small Animal Models of Enteric Diseases, NIAID, NIH, September 13-14, 2012. She also gave an invited talk entitled C. jejuni induces mixed Type 1 and 17 responses in acute and chronic disease for the Enterics Research Investigational Network, Cooperative Research Centers Annual Meeting, NIH, Seattle, Washington, May 22-23, 2012. North Dakota North Dakota State University (NDSU) and Makerere University (MAK) in Kampala, Uganda offered the joint course International Animal Production, Disease Surveillance and Public Health again in summer 2012 with 23 student attendees  5 from US and 18 from 6 higher education institutions in East Africa. Additionally, 6 students from MAK enrolled in the MS degree in International Infectious Disease Management attended the summer course. Also in July, 2012 an International conference was held in Kampala Uganda funded by USAID initiative on Africa-US Capacity building in Higher Education for Developmentthe project is Capacity building in Integrated Management of Transboundary Animal Diseases & Zoonoses in East & Central Africa (ECA). This project is currently implemented using One Health approach through AFRUS-IDM (Africa-US Integrated Disease Management), a network of higher education institutions in East & Central Africa (University of Nairobi, Kenya; Sokoine University of Agriculture, Tanzania; Mekelle University, Ethiopia; Umutara Polytechnic, Rwanda; Sheik Technical Veterinary School, Somalia) and the US and Canada (Columbus State University, Kansas State University, Michigan State University, University of Minnesota, Washington State University, and University of Saskatchewan, Canada), respectively. Nebraska Knowledge pertinent to NC-1041 activities was disseminated to undergraduate students, graduate students, professional veterinary students, veterinarians, physicians, food processors, researchers, cattle producers and other decision makers regarding pre-harvest food safety of cattle food projects. A listing of specific presentations and dates is provided in Section VI, below.

1. Sahin O, Fitzgerald C, Stroika S, Zhao S, Sippy RJ, Kwan P, Plummer PJ, Han J, Yaeger MJ, Zhang Q. 2012. Molecular Evidence for Zoonotic Transmission of an Emergent Highly Pathogenic Campylobacter jejuni Clone in the United States. J. Clin. Microbiol. 50:680-687. PMID: 22189122. PMCID: PMC3295108. 2. Plummer P, Sahin O, Burrough E, Sippy R, Mou K, Rabenold J, Yaeger M, Zhang Q. 2012. Critical Role of LuxS in the Virulence of Campylobacter jejuni in a Guinea Pig Model of Abortion. Infect. Immun. 80:585-93. PMID: 22144479. PMCID: PMC3264297. 3. Luo Y, Sahin O, Dai L, Sippy R, Wu Z, Zhang Q. 2012. Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive and Specific Detection of a Campylobacter jejuni Clone. J. Vet. Med. Sci. 74:591596. PMID: 22188995. 4. Burrough ER, Zuowei Wu, Orhan Sahin, Zhang Qijing, Mike J. Yaeger. 2012. Spatial Distribution of Putative Growth Factors in the Guinea Pig Placenta and the Effects of These Factors, Plasma, and Bile on the Growth and Chemotaxis of C. jejuni. Vet. Pathol. 49:470-481. PMID: 22081135. 5. Luangtongkum T, Shen Z, Seng VW, Sahin O, Jeon B, Liu P, Zhang Q. 2012. Impaired fitness and transmission of macrolide-resistant Campylobacter jejuni in its natural host. Antimicrob. Agents Chemother. 56:1300-1308. PMID: 22183170. PMCID: PMC3294946. 6. Han J, Wang Y, Sahin O, Shen Z, Guo B, Shen J, and Zhang Q. 2012. A fluoroquinolone resistance associated mutation in gyrA affects DNA supercoiling in Campylobacter jejuni. Front. Microbiol. 2:21. PMID: 22919613. PMCID: PMC3417464. 7. Sippy R, Sandoval-Green CM, Sahin O, Plummer P, Fairbanks WS, Zhang Q, Blanchong JA. 2012. Occurrence and molecular analysis of Campylobacter in wildlife on livestock farms. Vet. Micro. 157:369-75. PMID: 22266157. 8. Hwang S, Zhang Q, Ryu S, Jeon B. 2012. Transcriptional regulation of the CmeABC multidrug efflux pump and the KatA catalase by CosR in Campylobacter jejuni. J Bacteriol. 2012 Oct 12. [Epub ahead of print]. PMID: 23065977. 9. Shen Z, Pu XY, and Zhang Q. 2011. Salicylate functions as an efflux pump inducer and promotes the emergence of fluoroquinolone-resistant Campylobacter jejuni mutants. Appl Environ Microbiol. 77:7128-33. PMID: 21821741. PMCID: PMC3194847. 10. Kassem II, Sanad YM, Stonerock R, Rajashekara G. 2012. An evaluation of the effect of sodium bisulfate as a feed additive on Salmonella enterica serotype Enteritidis in experimentally infected broilers. Poult Sci. Apr;91(4):1032-7. 11. Yasser M. Sanad, Gary Closs Jr, Anand Kumar, Jeffrey T. LeJeune, Gireesh Rajashekara. 2012. Molecular Epidemiology and Public Health Relevance of Campylobacter Isolated from Dairy Cattle and European Starlings in Ohio, USA. Foodborne pathogens and diseases, (In press) 12. Issmat I. Kassem1, Mahesh Khatri, Malak A. Esseili, Yasser M. Sanad, Yehia M. Saif, Jonathan W. Olson2, Gireesh Rajashekara. 2012. Respiratory proteins contribute differentially to Campylobacter jejunis survival and in vitro interaction with hosts intestinal cells. BMC Microbiology, Nov 13;12(1):258. 13. Sanad, Y.M., I.I. Kassem, Z. Liu, J. Lin, J. T. Lejune, and G. Rajashekara. 2012. Occurrence of the invasion associated marker (iam) in C. jejuni isolated from cattle. BMC Research Notes. 4:570 14. Jun Lin J, KS Mateo, M Zhao, AK Erickson, N Garcia, D He, RA Moxley and DH Francis. 2012. Protection of Piglets against Enteric Colibacillosis by Intranasal Immunization with K88ac (F4ac) Fimbriae and Heat Labile Enterotoxin of Escherichia coli. Vet Microbiol. Epub ahead of press. 15. Zeng, X., F. Xu, Y. Mo, and J. Lin. 2012. Identification and characterization of a periplasmic trilactone esterase, Cee, revealed novel features of ferric enterobactin acquisition in Campylobacter. Molecular Microbiology (Accepted). 16. Theoret JR, Cooper KK, Zekarias B, Roland KL, Law BF, Curtiss R 3rd, Joens LA. The Campylobacter jejuni Dps Homologue Is Important for In Vitro Biofilm Formation and Cecal Colonization of Poultry and May Serve as a Protective Antigen for Vaccination. Clin Vaccine Immunol. 2012 ;19(9):1426-31. 17. Cooper KK, Cooper MA, Zuccolo A, Joens LA. Re-sequencing of a virulent strain of Campylobacter jejuni NCTC11168 reveals potential virulence factors. Res Microbiol. 2012 Oct 6. doi:pii: S0923-2508(12)00136-2. 10.1016/j.resmic.2012.10.002. [Epub ahead of print] 18. Bell JA, Jerome JP, Plovanich-Jones AE, Smith EJ, Gettings JR, Kim HY, Landgraf JR, Lefébure T, Kopper JJ, Rathinam VA, St Charles JL, Buffa BA, Brooks AP, Poe SA, Eaton KA, Stanhope MJ, Mansfield LS. Outcome of infection of C57BL/6 IL-10(-/-) mice with Campylobacter jejuni strains is correlated with genome content of open reading frames up- and down-regulated in vivo. Microbial Pathogenesis 2012 Aug 31. pii: S0882-4010(12)00151-9. doi: 10.1016/j.micpath.2012.08.001. 19. Jerome J.P., Klahn B.D., Bell J.A., Barrick J.E., Brown C.T., and Mansfield L.S. 2012 Draft Genome Sequences of Two C. jejuni Clinical Isolates, NW and D2600, Journal of Bacteriology, in press. 20. Kim JS, Artymovich KA, Hall DF, Smith EJ, Fulton R, Bell JA, Dybas L, Mansfield LS, Tempelman R, Wilson DL, Linz JE. 2012. Passage of Campylobacter jejuni through the chicken reservoir or mice promotes phase variation in contingency genes Cj0045 and Cj0170 that strongly associates with 21. colonization and disease in a mouse model. Microbiology February 16, 2012 as doi:10.1099/mic.0.057158-0. 22. Vannucci FA, Beckler D, Pusterla N, Mapes SM, Gebhart CJ. 2012. Attenuation of virulence of Lawsonia intracellularis after in vitro passages and its effects on the experimental reproduction of porcine proliferative enteropathy. Vet. Micro. (In press) 23. Sampieri F, Vannucci FA, Allen AL, Pusterla N, Antonopoulos A.J., Ball K.R., Thompson J., Dowling P.M., Hamilton D.L., Gebhart C.J. 2012. Species-specificity of equine and porcine L. intracellularis isolates in laboratory animals. Can J Vet Res. (In press) 24. Chander Y, Primus A, Oliveira S, Gebhart CJ. 2012. Phenotypic and molecular characterization of a novel strongly hemolytic Brachyspira species, provisionally designated Brachyspira hampsonii J. Vet. Diag. Investi. 24:903-910. 25. Pusterla N, Mapes S, Gebhart C. 2012. L. intracellularis-specific interferon c gene expression by peripheral blood mononuclear cells in vaccinated and naturally infected foals. Vet. J. 192:249-251. 26. Sampieri F, Allen AL, Pusterla N, Vannucci FA, Anatonopoulos AJ, Hamilton, DL, and Gebhart CJ. 2012. The rabbit as an infection model for equine proliferative enteropathy. Can J. Vet. Res. (In press ) 27. Viott AM, Franca SA, Vannucci FA, Cruz ECC, Costa MC, Gebhart CJ and Guedes MC. 2012. Infection of sparrows (Passer domesticus) and different mice strains with Lawsonia intracellularis. Pesquisa Veterinaria Brazileira. (In press) 28. Vannucci, FA, Foster DN and Gebhart CJ. 2012. Comparative transcriptional analysis of homologous pathogenic and non-pathogenic Lawsonia intracellularis isolates in infected porcine cells. PloS ONE 7(10): e46708. doi:10.1371/journal.pone.0046708 29. Nogradi N, Slovis NM, Gebhart CJ, Wolfsdorf KE, McCrcken JL, Scoggin CF, Kass PH, Mapes SM, Toth B, Lundquist ML, Pusterla N. 2012. Evaluation of the field efficacy of an avirulent live Lawsonia intracellularis vaccine in foals. Vet. J. 192:511-513. 30. Pusterla N, Vannucci FA, Guzman DSM, Mapes S, White A, DiFrancesco M, Gebhart C. 2012. Transmission of Lawsonia intracellularis to weanling foals using feces from experimentally infected rabbits. Vet. Journal. (In press). 31. Vannucci FA, Pusterla N, Mapes SM, Kelley M, Gebhart CJ. 2012. Evidence of host adaptation in Lawsonia intracellularis infections. Vet Res. 43:53-65. 32. Pusterla N., Mapes S. and Gebhart C. 2012. Further investigation of exposure to Lawsonia intracellularis in wild and feral animals captured on horse properties with equine proliferative enteropathy. Vet. J. 194:253-255. 33. Vannucci F.A., Wattanaphansak S., Gebhart C.J. 2012. An alternative method for cultivation of Lawsonia intracellularis. J. Clin. Microbiol. 50:1070-1072. 34. Pusterla N., Vannucci, F.A., Mapes S.M., Nogradi, N., Collier J.R., Hill J.A., DiFrancesco M., White A.M., Akana N.K., Simonek G., Gebhart C.J. 2012. Efficacy of an avirulent live vaccine against Lawsonia intracellularis in the prevention of proliferative enteropathy in experimentally infected weanling foals. Am. Jr. Vet. Res. 73:741-746. 35. Paradis M., Gebhart C.J., Toole D., Vessie G., Winkelman N.L., Bauer S.A., Wilson J.F., McClure C.A. 2012. Subclinical Ileitis: Diagnostic and performance parameters in a multi-dose mucosal homogenate challenge model. J. Swine Health and Prod. 20:137-141. 36. Cull CA, Paddock ZD, Nagaraja TG, Bello NB, Babcock AH, Renter DG. Efficacy of a vaccine and a direct-fed microbial against fecal shedding of Escherichia coli O157:H7 in a randomized pen-level field trial of commercial feedlot cattle. Vaccine 2012; 30(43): 6210-6205. 37. Jacob ME, Nagaraja TG. Use of direct-fed microbials as a preharvest food safety intervention in cattle. p 189-202. In: Direct-Fed Microbials and Probiotics for Animals: Science and Mechanisms of Action. T. R. Callaway and S. C. Ricke (eds.) 2012. Springer Publ., NY. 38. Bai J, Paddock ZD, Shi X, Li S, An B, Nagaraja TG.. Applicability of a Multiplex PCR to detect the seven major Shiga toxin-producing Escherichia coli based on genes that code for serogroup-specific O-antigens and major virulence factors in cattle feces. Foodborne Path Dis. 2012, 9:541-548. 39. Paddock ZD, Shi X, Bai J, Nagaraja TG. Applicability of a multiplex PCR to detect O26, O45, O103, O111, O121, O145, and O157 serogroups of Escherichia coli in cattle feces. Vet Microbiol 2012, 156:381-388. 40. Jacob ME, Shi X, An B, Nagaraja TG, Bai J. Evaluation of a multiplex real-time PCR for the quantification of Escherichia coli O157 in cattle feces. Foodborne Path Dis 2012, 9:79-85. 41. Eberhart LJ, Deringer JR, Brayton KA, Sawant AA, Besser TE, Call DR. Characterization of a novel microcin that kills enterohemorrhagic Escherichia coli O157:H7 and O26. Appl Environ Microbiol. 2012 Sep;78(18):6592-9. doi: 10.1128/AEM.01067-12. PMID: 22773653; PMCID: PMC3426703. 42. Irshad H, Cookson AL, Hotter G, Besser TE, On SL, French NP. Epidemiology of Shiga toxin-producing Escherichia coli O157 in very young calves in the North Island of New Zealand. N Z Vet J. 2012 Jan;60(1):21-6. PubMed PMID: 22175425. 43. Kudva IT, Davis MA, Griffin RW, Garren J, Murray M, John M, Hovde CJ, Calderwood SB. Polymorphic Amplified Typing Sequences and Pulsed-Field GelElectrophoresis Yield Comparable Results in the Strain Typing of a Diverse Set of Bovine Escherichia coli O157:H7 Isolates. Int J Microbiol. 2012;2012:140105. doi: 10.1155/2012/140105. Epub 2012 Aug 7. PubMed PMID: 23049559; PubMed CentralPMCID: PMC3461292. 44. Bono JL, Smith TP, Keen JE, Harhay GP, McDaneld TG, Mandrell RE, Jung WK, Besser TE, Gerner-Smidt P, Bielaszewska M, Karch H, Clawson ML. Phylogeny of Shiga toxin-producing Escherichia coli O157 isolated from cattle and clinically ill humans. Mol Biol Evol. 2012 Aug;29(8):2047-62. Epub 2012 Feb 21. PubMed PMID: 22355013; PubMed Central PMCID: PMC3408066. 45. Shringi S, García A, Lahmers KK, Potter KA, Muthupalani S, Swennes AG, Hovde CJ, Call DR, Fox JG, Besser TE. Differential virulence of clinical and bovine-biased enterohemorrhagic Escherichia coli O157:H7 genotypes in piglet and Dutch belted rabbit models. Infect Immun. 2012 Jan;80(1):369-80. PMID: 22025512; PMCID: PMC3255674. 46. Pham T, Gao X, Tsai K, Olsen R, Wan F, Hardwidge PR. Functional differences and interactions between the E. coli type III secretion system effectors NleH1 and NleH2, Infection & Immunity, 2012 Jun;80(6):2133-40. PMID: 22451523 47. Wang X, Hardwidge, PR. Enterotoxigenic Escherichia coli (ETEC) prevents host NF-kB activation by targeting IkBa polyubiquitination, Infection & Immunity, 2012, Oct.1 [Epub ahead of print] PMID: 23027537 48. Wang X, Gao X, Hardwidge PR. Heat-labile enterotoxin-induced activation of NF-kappaB and MAPK pathways in intestinal epithelial cells impacts enterotoxigenic Escherichia coli (ETEC) adherence, Cellular Microbiology, 2012 Aug;14(8):1231-41. PMID: 22452361. 49. Zhang, W. & D. A. Sack. 2012. Progress and hurdles in the development of vaccines against enterotoxigenic Escherichia coli in humans. Expert Review of Vaccines. 11(6):677-94. 50. Ruan, X., Crupper S.C., Schultz B.D., Robertson D.C., and W. Zhang. 2012. Escherichia coli expressing enteroaggregative heat-stable toxin 1 (EAST1) did not cause an increase of cAMP or cGMP levels in cells, and no diarrhea in 5-day old gnotobiotic pigs. PLoS ONE. 7(8):e43203. 51. Duan, Q, M. Zhou, X. Zhu, Y. Yang, J. Zhu, W. Bao, S. Wu, X. Ruan, W. Zhang, and Zhu G. 2012. Flagella from F18+ Escherichia coli play a role in adhesion to pig epithelial cell lines. Mcrobiol Pathogen. In press. 52. Duan, Q., Zhou M., Zhu X., Bao W., Wu S., Ruan X., Zhang W., Yang Y., Zhu J., and Zhu G. 2012. The flagella of F18ab Escherichia coli is a virulence factor that contributes to infection in an IPEC-J2 cell model in vitro. Vet. Microbiol. 160(2012):132-140. 53. Santiago-Mateo K, M Zhao, J Lin, W. Zhang, DH Francis. 2012. Avirulent K88 (F4)+ Escherichia coli strains constructed to express modified enterotoxins protect young piglets from challenge with a virulent enterotoxigenic E. coli strain that expresses the same adhesion and enterotoxins. Vet Microbiol.159:337-342. 54. Erume J, P Wijemanne, EM Berberov SD. Kachman, DJ Oestmann, DH. Francis, RA. Moxley. 2012. Inverse relationship between heat stable enterotoxin-b induced fluid accumulation and adherence of F4ac-positive enterotoxigenic Escherichia coli in ligated jejunal loops of F4ab/ac fimbria receptor-positive swine. Vet Microbiol 161:315-324. 55. Kim Y, Lovell S, Tiew K, Mandadapu SR, Alliston KR, Battaile KP, Groutas WC, Chang K. Novel Broad-Spectrum Antivirals against 3C or 3C-like proteases of Picornaviruses, Noroviruses and Coronaviruses. Journal of Virology, 2012, 86(21):11754-62. 56. Kim Y, George D, Prior AM, Prasain K, Hao S, Le DD, Hua DH, Chang K. Novel triacsin C analogs as potential antivirals against rotavirus infections. Eur J Med Chem, 2012, 50:311-8. 57. Esseili MA, Wang Q, Zhang Z, Saif LJ. Internalization of sapovirus, a surrogate for norovirus, in romaine lettuce and the effect of lettuce latex on virus infectivity. Appl Environ Microbiol. 2012. 78:6271-9 58. Esseili MA, Wang Q, Saif LJ. 2012. Binding of human GII.4 norovirus virus-like particles to carbohydrates of romaine lettuce leaf cell wall materials. Appl Environ Microbiol. 78:786-94. 59. Jung, K., Q. Wang, Y. Kim, K. Scheuer, Z. Zhang, Q. Shen, K-O. Chang, and L. J. Saif. 2012. The effects of simvastatin or interferon-± on infectivity of human norovirus using a gnotobiotic pig model for the study of antivirals. PLoS One 7:e41619. 60. Wang, Q., K. Scheuer, Z. Ahang, W. A. Gebreyes, B. Z. Molla, A. E. Hoet, and L. J. Saif. 2011. Characterization and prevalence of a new porcine Calicivirus in Swine, United States. Emerging infectious diseases 17:1103-1106. 61. Wang Q., Z. Zhang and L.J. Saif. 2012. A cultivable porcine sapovirus surrogate for human caliciviruses: stability and attachment to lettuce. Appl Environ Microbiol. 78:3932-40. 62. Harada S., Oka T., Tokuoka E., Kiyota N., Nishimura K., Shimada Y., Ueno T., Ikezawa S., Wakita T., Wang Q. Saif L. J., Katayama K. 2012. A confirmation of sapovirus re-infection gastroenteritis caseswith different genogroups and genetic shifts in the evolving sapovirus genotypes, 2002-2011. Arch Virol 157:19992003. 63. Amimo, J.O., Vlasova, A.N., Saif, L.J. 2013. Prevalence, genetic heterogeneity and identification of a potential new VP4 genotype of porcine group c rotaviruses in nursing and weaned piglets in Ohio, USA. Submitted to Veterinary Microbiology. 64. Khaitsa, M.L. and Dawn Doetkott. 2012 (Accepted in August 2011). Antimicrobial Drug Resistance and Molecular Characterization of Salmonella Isolated from Domestic Animals, Humans and Meat Products, In: Salmonella - A Dangerous Foodborne Pathogen, Barakat S. M. Mahmoud (Ed.), ISBN: 978-953-307-782-6. 65. Michael Muleme, Robert Barigye, Margaret L. Khaitsa, Eugene Berry, Anthony W. Wamono, Chrisostom Ayebazibwe. 2012. Effectiveness of vaccines and vaccination programs for the control of foot-and-mouth disease in Uganda, 20012010. Trop. Ani. Health and Prod. September 2012. 66. Stella Opendi Sasanya, Susan Olet, Robert Littlefield, Charles L. Stoltenow and Margaret L. Khaitsa. 2012. Spatial-temporal Distribution of the 2007 Melamine Associated Nephrotoxic Renal failure among Pets and Factors Associated with Pet Survival. Journal of Food Protection Trends. In Press. 67. George S, K Circle, S Lindblom, S Vilain, AJM Rosa, D Francis, V Brözel1, and RS Kaushik. 2012. Assessment of Toll-like Receptors in the Ileum of Weanling Pigs- Responses to Feed Antibiotic Chlortetracycline and Gnotobiotic Conditions. Clin Cellular Immunol. Epub ahead of print. 68. Wang, Z., X. Zeng, Y. Mo, K. Smith, J. Lin. 2012. Identification and characterization of a bile salt hydrolase for developing novel alternatives to antibiotic growth promoters. Applied and Environmental Microbiology 78:8795-8802 69. Lin, J., A.A. Hunkapillar, A.C. Layton, Y. Chang, K.R. Robbins. 2012. Response of intestinal microbiota to antibiotic growth promoters in chickens. Foodborne Pathogens and Diseases (In Press) 70. Hoang, K., Y. Wang, and J. Lin. 2012. Identification of genetic loci required for Campylobacter resistance to fowlicidin-1, a chicken host defense peptide. Frontiers in Cellular and Infection Microbiology 2:32. Doi:10.3389/fcimb.2012.00032 71. Zhou, W., Y. Wang, and J. Lin. 2012. Functional cloning and characterization of antibiotic resistance genes from chicken gut microbiome. Applied and Environmental Microbiology. 78:3028-3032 72. Cummings KJ*, Warnick LD, Davis MA, Eckmann K, Gröhn YT, Hoelzer K, MacDonald JK, Root TP, Siler JD, McGuire SM, Wiedmann M, Wright EM, Zansky SM, Besser TE. Farm animal contact is a risk factor for salmonellosis caused by bovine-associated subtypes. (Accepted, Emerg Inf Dis). 73. Morales CA, Guard J, Sanchez-Ingunza R, Shah DH, Harrison M. Virulence and metabolic characteristics of Salmonella enterica serovar enteritidis strains with different sefD variants in hens. Appl Environ Microbiol. 2012 Sep;78(18):6405-12. doi: 10.1128/AEM.00852-12. Epub 2012 Jun 22. PubMed PMID: 22729535; PubMed Central PMCID: PMC3426706. 74. Neal-McKinney JM, Lu X, Duong T, Larson CL, Call DR, Shah DH, Konkel ME. Production of organic acids by probiotic lactobacilli can be used to reduce pathogen load in poultry. PLoS One. 2012;7(9):e43928. doi: 10.1371/journal.pone.0043928. Epub 2012 Sep 4. PMID: 22962594; PMCID: PMC3433458. 75. Shah DH, Casavant C, Hawley Q, Addwebi T, Call DR, Guard J. Salmonella Enteritidis strains from poultry exhibit differential responses to acid stress, oxidative stress, and survival in the egg albumen. Foodborne Pathog Dis. 2012 Mar;9(3):258-64. PMID: 22304629; PMCID: PMC3326446. 76. Shah DH, Zhou X, Kim HY, Call DR, Guard J. Transposon Mutagenesis of Salmonella enterica Serovar Enteritidis Identifies Genes That Contribute to Invasiveness in Human and Chicken Cells and Survival in Egg Albumen. Infect Immun. 2012 Dec;80(12):4203-15. PMID: 22988017; PMCID: PMC3497420. 77. Subbiah M, Shah DH, Besser TE, Ullman JL, Call DR. Urine from Treated Cattle Drives Selection for Cephalosporin Resistant Escherichia coli in Soil. PLoS One. 2012;7(11):e48919. doi: 10.1371/journal.pone.0048919. PMID: 23145021; PMCID: PMC3492140. 78. Morrison CM, Dial SM, Day WA Jr, Joens LA. Investigations of Salmonella enterica serovar newport infections of oysters by using immunohistochemistry and knockout mutagenesis. Appl Environ Microbiol. 2012 Apr;78(8):2867-73. Epub 2012 Feb 3. 79. Li M, Bauer LL, Chen X, Wang M, Kuhlenschmidt TB, Kuhlenschmidt MS, Fahey GC, Jr, and Donovan SM (2012) Microbial Composition and In Vitro Oligosaccharides and Prebiotics Differ between Formula-Fed and Sow-Reared Piglets. J. Nutrition 142:1-10. 80. Fuller KL, Kuhlenschmidt TB, Kuhlenschmidt MS, Jimenez-Flores R, and Donovan SM. (2012) Milk Fat Globule Membrane Isolated from Buttermilk or Cheese Whey and their Lipid Component Inhibit Infectivity of Rotavirus In Vitro. J. Dairy Science (in press). 81. Li, M, Monaco MH, Wang, M, Comstock SS, Kuhlenschmidt TB, Kuhlenschmidt MS, Donovan SM, (2013) Human milk oligosaccharides and prebiotics reduce the duration of rotavirus-induced diarrhea in piglets. JPGN (in press). 82. Lin, J., K. S. Mateo, M. Zhao, A. K. Erickson, N. Garcia, D. He, R. A. Moxley, and D. H. Francis. 2012. Protection of piglets against enteric colibacillosis by intranasal immunization with K88ac (F4ac) fimbriae and heat labile enterotoxin of Escherichia coli. Vet. Microbiol. (in press, VETMIC-5932) 83. Shippy, D. C. and A. A. Fadl. 2012. Immunological characterization of a gidA mutant strain of Salmonella for potential use in a live-attenuated vaccine. BMC Microbiology (Accepted for publication).