Brief Summary of Minutes of Annual Meeting

1. Linda Mansfield, Chair of NC-1041, gave an update on the proposal renewal. The proposal has been finished and uploaded onto the web site for NC-1041. Anybody interested in the proposal can print out a copy directly from the web page. 2. Appendix E uploading. Members of NC-1041 are required to submit the Appendix E form. This must be done by Jan. 15, 2012 via the NIMSS system. Members dont have to be state representatives. Investigators from medical schools are allowed and encouraged to become member of NC-1041. 3. Qijing Zhang and Lynn Joens will become the Chair and Chair-Elect, respectively, at the 2012 annual meeting. The term will be for 2 years. 4. New members were introduced. 5. Student awards. NC-1041 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 suggested. 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-1041 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. 6. Proposed title change for the Gastroenteric Disease Section. There was a motion to change the section name from Gastroenteric Disease to Gastroenteric Disease, Zoonosis and Food Safety. The rationale for the change is that many members of NC-1041 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. 7. Annual report. Annual state reports are due to Linda Mansfield 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.

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 Emerging Campylobacter Species. The best-studied species, C. jejuni and C. coli, exhibit high genomic diversity. However, after sequencing more than forty strains from different hosts, Stanhope and colleagues showed that there are barriers to interspecies recombination in the core genomes of the two species. Additionally, phenotypic differences in disease produced can be discerned among strains of C. jejuni. We delineated five pathotypes of C. jejuni in C57BL/6 IL-10-/- mice: lack of colonization, colonization without disease, colonization with watery or hemorrhagic enteritis, and colonization with neurological signs with or without enteritis. These pathotypes correspond to the spectrum of human disease; however, gene content did not predict pathotype: each strain possessed a unique set of present, absent or divergent virulence factors. Tennessee C. jejun is the leading foodbome human pathogen in the United States and many other industrialized countries. Increasing evidence also indicates that antibiotic use in poultry selects for resistant C. jejuni, posing a significant threat to public health. We have made significant progress to understand the molecular mechanisms of pathogenesis and antibiotic resistance in Campylobacter during the reporting period. The findings have filled a significant gap in antimicrobial resistance development in Campylobacter and provided important information for the development of effective vaccine to control Campylobacter. Iowa A highly virulent, tetracycline-resistant C. jejuni clone (clone SA) has recently emerged in ruminant reservoirs and has become the predominant cause of sheep abortion in the United States. To determine whether clone SA is associated with human disease, we compared the clinical isolates of clone SA from sheep abortions with the human isolates of the PulseNet National Campylobacter databases at the CDC and the FDA using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and serotyping. The results provide strong molecular and epidemiological evidence for zoonotic transmission of this emergent clone from ruminants to humans and indicate that C. jejuni clone SA is an important threat to public health. Additionally, we develop a LAMP assay specific for detection of clone SA of Campylobacter jejuni. In another study, we determined the direct role of luxS in the virulence of C. jejuni in two different animal hosts. The IA3902 strain, a highly virulent sheep abortion strain recently described by our laboratory, along with its isogenic luxS mutant and luxS complement strains, was inoculated by the oral route into both a pregnant guinea pig virulence model and a chicken colonization model. In both cases, the IA3902 luxS mutant demonstrated a complete loss of ability to colonize the intestinal tract. In the pregnant model, the mutant also failed to induce abortion, while the wild-type strain was highly abortifacient. Genetic complementation of the luxS gene fully restored the virulent phenotype in both models. Interestingly, when the organism was inoculated into guinea pigs by the intraperitoneal route, no difference in virulence (abortion induction) was observed between the luxS mutant and the wild-type strain, suggesting that the defect in virulence following oral inoculation is likely associated with a defect in colonization and/or translocation of the organism out of the intestine. These studies provide the first direct evidence that LuxS plays an important role in the virulence of C. jejuni using an in vivo model of natural disease. B. Non-O157 Shiga toxin-producing E. coli (STEC) Nebraska Validation of Culture Methods for Non-O157 STEC. Non-O157 STEC are enzootic in cattle and constitute an emerging zoonotic threat. A current research goal of the USDA-NIFA is the development of diagnostic tests for STEC O26, O45, O103, O111, O121, O145, and O157:H7. Preliminary studies were conducted to test the validity of a published culture protocol (Possé et al. 2008. FEMS Microbiol. Lett. 282:124-131) for detection of STEC O26, O103, O111, O145, and O157:H7, and to determine whether it can be used for detection of O45 and O121. The results suggest that the differential culture medium described by Possé et al. (FEMS Microbiol. Lett. 2008;282:124-131) can be used to concurrently screen for all seven serogroups (O26, O45, O103, O111, O121, O145, and O157:H7). More research on the development and validation of detection methods for non-O157 STEC is needed. North Dakota A study was conducted to investigate prevalence of Shigatoxin-producing Escherichia coli (STEC) in beef cattle in North Dakota at different production stages from post-weaning to slaughter & testing for presence of Shiga toxin 1 (stx1), Shiga toxin 2 (stx2), Intimin (eaeA) and Enterohemolysin (ehlyA) genes using PCR. The preliminary data indicate presence of several serotypes of Shigatoxin-producing Escherichia coli (STEC) (O157, 0145, 0111, 0103, 026) in feces of beef cattle (both calves and adult cattle) in ND. Prevalence of Shiga toxin-producing Escherichia coli serogroup O26 in feces of feedlot cattle. Our preliminary findings are that O26 are common, but with a variable prevalence, in cohorts of feedlot cattle. Furthermore, fecal prevalence of O26 and presence of virulence genes may be less than what was observed for serogroup O157. However, the fecal prevalence of O26 may have a similar temporal distribution in feedlot cattle as observed for O157. C. Caliciviruses Ohio The binding mechanisms of HuNoV to leafy greens. Our results indicate that NoV VLPs bind to lettuce CWM specifically by utilizing multiple carbohydrate moieties. This binding may enhance virus persistence on the leaf surface and thus prevent effective removal by simple washing and disinfection by routine decontamination procedures. These results should help to develop more effective decontamination procedures to control foodborne HuNoV outbreaks. Persistence and mode of transmission of sapovirus, a surrogate for norovirus, in leafy greens. Our results indicate that (i) SaV can potentially be transferred from roots to leaves and (ii) that the virus can persist for a prolonged period on the leaf surface once introduced. The persistence of the virus in/on lettuce and the possibility of contamination through roots suggest the need for more rigorous pre-harvest measures to reduce calicivirus contamination. Potential interspecies transmission of NoVs between pigs and humans. Although no HuNoVs were detected from swine farms, our data is limited to only one state, North Carolina. Additional studies of porcine NoVs from other states are needed to assess their geographic and age prevalence in pigs and to confirm their relatedness to human strains. Detection and characterization of the first feline NoV. Our findings document the discovery of a novel feline calicivirus, different from vesiviruses, and extend the spectrum of NoV host range. Epidemiological studies using feline NoV-specific diagnostic tools and experimental infection of cats are required to understand whether NoVs have a pathogenic role in this species or may be transmitted to humans. Regulations may be needed to control the spread of such new viruses. Kansas Characterization and inhibition of norovirus proteases of genogroups I and II using a fluorescence resonance energy transfer assay. Noroviruses are the major cause of food- or water-borne gastroenteritis outbreaks in humans. The norovirus protease that cleaves a large viral polyprotein to nonstructural proteins is essential for virus replication and an attractive target for antiviral drug development. Noroviruses show high genetic diversity with at least five genogroups, GI-GV, of which GI and GII are responsible for the majority of norovirus infections in humans. We cloned and expressed proteases of Norwalk virus (GI) and MD145 virus (GII) and characterized the enzymatic activities with fluorescence resonance energy transfer (Fret) substrates. We have established a high throughput assay for screening potential protease inhibitors using the Fret assay. We also demonstrated that the GI and GII proteases cleaved the substrates derived from the naturally occurring cleavage site in the open reading frame (ORF) 1 of G1 norovirus with similar efficiency, and that enzymatic activity of both proteases were inhibited by commercial protease inhibitors including chymostatin. The interaction of chymostatin to Norwalk virus protease was validated by nuclear magnetic resonance (NMR) spectroscopy. The established high throughput assay would be highly useful for screening potential antivirals and study of enzyme-substrate interactions. D. Rotavirus Illinois Our findings demonstrate a dramatic reduction in virus shedding and absence of rotavirus disease in SLPE-fed piglets, thus providing proof of concept that a nutriceutical approach of using natural or synthetic receptor mimetics, such as SLPE, hold substantial promise for the prevention of rotavirus disease. This approach may be applicable not only to piglets in a natural field setting, but also as a potential dietary supplement, for example in infant formulae, for the prevention of rotavirus or other gastrointestinal infectious diseases in human infants. E. Enterococci Kansas Selection of tcrB gene mediated copper resistant fecal enterococci in pigs fed diets supplemented with copper. The higher prevalence of tcrB-positive enterococci in piglets fed elevated copper compared to normal copper suggests that supplementation of copper in swine diets selected for resistance. The existence of a metalassociated co-selection mechanism could potentially be a major issue relative to public health and further studies are needed to determine the magnitude of such an association. Characterization of antibiotic resistant and virulent enterococci from animal feed. Our data revealed that feed samples collected from feed mills and swine farms carried antibiotic resistant and potentially virulent enterococci. We have shown that these isolates have the potential to spread the antibiotic resistance by horizontal gene transfer. Insects in confined swine operations carry a large antibiotic resistant and potentially virulent enterococcal community. This study shows that house flies and German cockroaches in the confined swine production environment likely serve as vectors and/or reservoirs of antibiotic resistant and potentially virulent enterococci and consequently may play an important role in animal and public health. F. E. coli O157 Kansas 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. Our data, although based on a limited number of samples, suggest that the sensitivities of the mPCR and culture-based methods in detecting the seven serogroups of STEC in feces differed between O-groups. An obvious limitation of our mPCR is that the concurrent detection of virulence genes and the serogroups in a sample does not necessarily associate the virulence genes with the prevalent serogroups in the same sample. The major application of our11-gene mPCR assay may be in identifying putative colonies of STEC obtained by culture-based methods. Escherichia coli O157:H7 genetic diversity in bovine fecal samples. A total of 254 E. coli O157:H7 isolates from 27 samples were included in PFGE analysis. Fifteen PFGE subtypes (< 100% Dice similarity) were detected among the 254 isolates and there were no common subtypes between the three locations. Seven of 27 (26%) fecal samples had E. coli O157:H7 isolates with different PFGE subtypes (mean = 2.1) within the same sample. The virulence gene profiles of different isolates from the same sample were always identical, regardless of the number of PFGE types. The results of this study suggest that determining the PFGE pattern of a single isolate from a bovine sample may not be sufficient when comparing to other isolates from feces, hides or carcasses as multiple PFGE subtypes are present. G. Lawsonia intracellularis Minnesota Subclinical porcine proliferative enteropathy: diagnostic and performance parameters in a multi-dose mucosal homogenate challenge model. The objective of this study was to further define subclinical PE by measuring the impact of varying doses of Lawsonia on clinical signs, Lawsonia shedding and seropositivity, performance, and gross and histopathological intestinal changes in weaned pigs. The results showed that subclinical infection can have a detrimental economical disease impact on swine herds. This study confirms that it is possible to reproduce a spectrum of PE that includes mild and subclinical disease using a mucosal homogenate challenge model which mimics naturally occurring disease in the field. Loop mediated isothermal amplification method for detection of L. intracellularis. We have developed a new diagnostic method, based on loop mediated isothermal amplification (LAMP), for the detection of L. intracellularis DNA in fecal samples. Using this method we were able to detect about 10,000 in 1 ml of L. intracellularis. No amplification was obtained with DNA extracts from other swine pathogens, indicating high specificity of this new method. Furthermore, this method was used for detection of L. intracellularis in fecal samples without any extensive DNA extraction process or the use of thermocyclers. 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 Using a mouse model of campylobacteriosis we have shown that C. jejuni NCTC11168 re-isolated from infected mice (mouse-adapted) is more virulent than the ancestral inoculum. Mouse-adapted C. jejuni also had altered ability to autoagglutinate and interact with epithelial cells. After extensive genetic analysis including Illumina genome re-sequencing, we showed that the only genetic difference between mouse-adapted and ancestral C. jejuni was in homopolymeric guanine tracts of thirteen (out of twenty-nine) contingency loci. This suggests that contingency loci play a role in pathogenesis in the mouse model. We then tested the effect of bottlenecking contingency loci mutational diversity on C. jejuni virulence phenotypes. A variant subjected to a single colony bottleneck could not colonize mice. Campylobacter jejuni NCTC 11168 induces a mixed Type1 and Type17 cytokine and cellular response in IL-10 deficient murine host. The inflammatory process was characterized by analyzing for inflammatory cytokines and cell types in the colon and mesenteric lymph nodes, and by C. jejuni reactive antibody induction and cytokine response in serum. Colon homogenate ELISA revealed significant up regulation of IFN-³, IL-17A, IL-22, IL-6 and IL-1² but not IL-4. Mesenteric lymph node analysis revealed increases in IFN-³ positive cells in the CD4+, CD8+ and ³´ T cell and NK cell compartments. IL-17 positive cells were increased in CD4+ T cell and Thy1hiCD3- innate lymphocyte compartments. IL-22 positive cells were increased from CD4+ T cell and NK cell compartments. Time-dependent up regulation in C. jejuni specific IgG2b (Type 17 associated), IgG2c and IgG3 (Type1 associated), but not IgG1 (Type 2 associated) or IgM were also observed in the serum of infected mice. Addition of a Thelper-2 (Th2) immunomodulator to antibiotic treatment improves survival and decreases clinical signs of inflammatory bowel disease induced by Campylobacter jejuni infection in a murine model. We have also developed a mouse model of autoimmune diseases arising after C. jejuni infection including Inflammatory Bowel Disease and Guillain Barré Syndrome. Ohio Campylobacter in Beef Cattle. Our studies on cattle associated Campylobacter highlight the importance of cattle as a potential reservoir for clinically important Campylobacter. A better understanding of prevalence, distribution, and molecular epidemiology of C. jejuni strains from beef cattle in the US will constitute a significant preharvest preparedness effort. The TAT system and Tat-dependent proteins. Screening for small molecule chemical inhibitors that specifically inhibit TAT translocation would lead to novel antibacterials for control of Campylobacter. Such approaches will be more practical for use in chickens because of the risk associated with the emergence of resistant bacteria with the use of antibiotics. Identification of proteins that are translocated through the TAT system will help elucidate the basis for the TAT system contribution to C. jejuni physiology and virulence. Furthermore, a comprehensive analysis of TAT-substrates may reveal other virulence or host adaptation proteins, permitting development of novel antimicrobial strategies. Polyphosphate kinase. No homologs of ppk1 and ppk2 have been identified in genomes of mammals and avian species, and thus both PPK1 and PPK2 can be used as potential anti-C. jejuni targets to reduce the prevalence of C. jejuni in poultry and other livestock species as well as humans. Understanding the role of C. jejuni PPKs on the expression of other genes involved in colonization and persistence is highly valuable to understand many aspects of host-microbe interactions in food producing animals and humans. Further, this work should greatly extend our fundamental knowledge on PPK regulation on various virulence genes in prokaryotes. Iowa To facilitate the control of macrolide-resistant Campylobacter, it is necessary to understand if macrolide resistance affects the fitness and transmission of Campylobacter in its natural host. In this study we conducted pairwise competitions and comingling experiments in chickens using clonally related and isogenic C. jejuni strains, which are either susceptible or resistant to erythromycin (Ery). The findings clearly indicate that acquisition of macrolide resistance impairs the fitness and transmission of Campylobacter in chickens, suggesting that the prevalence of macrolide-resistant C. jejuni will likely decrease in the absence of antibiotic selection pressure. Arizona Identification of C. jejuni proteins involved in host colonization. A major thrust of our laboratory is examining C. jejuni proteins for their role in the colonization of chicks. Genes expressing proteins that are secreted and have a surface orientation, or are up-regulated, or express known colonization factors are being mutated and examined for their role in gut colonization. The results indicate that the CjLAJ3 protein maybe an excellent vaccine candidate to reduce the numbers of C. jejuni in chickens. Vaccination of chicks with the attenuated Salmonella vector expressing the CjLAJ2 protein. The Salmonella strain used as the vector was developed by Curtiss (2005) and has three different mutations which results in its attenuation. Gene CjLAJ2, encoding a putative hemolysin protein, was inserted into an expression plasmid, cloned and amplified in a E. coli shuttle vector, extracted, and electroporated into the Salmonella vector. The trials demonstrated a significant reduction of C. jejuni in cecal contents of chicks vaccinated with the vector expressing the CjLAJ2 protein and challenged with the homologous strain. In addition, a 1-log reduction was present in chicks challenged with a heterologous strain. B. E. coli South Dakota South Dakota has developed a live vaccine for ETEC that prevents diarrhea in pigs within 24h of the time of vaccination. The vaccine seems to offer both immediate competitive exclusion from the pathogen and longer term protection by immunity. Nebraska Effect of culture media on secretion of heat-labile enterotoxin by porcine-origin enterotoxigenic Escherichia coli strains. This study was done to determine whether porcine WT ETEC isolates and recombinant LT constructs from a porcine isolate secrete LT and if so, to compare their LT secretion levels when grown in different culture media supportive of LT production and secretion. This study demonstrated that porcine WT strains secrete LT, albeit to a lesser level than the high LT-producing prototype human WT strain H10407. The level of LT secretion of porcine WT strains may vary depending on the growth media used. Porcine WT strains and a recombinant LT+ derivative of a porcine WT strain secreted higher levels of LT than did an LT+ clone in DH5±. The apparent reason for the lower level of LT secreted by DH5± is that it has a less efficient type II secretion system, which was been shown in a previous study. C. Cryptosporidium parvum Illinois The goal of our research is this area is to define the early mechanisms of Apicomplexa-host interactions and to identify new drug candidates that can block these interactions. We have developed a battery of complementary in vitro and in vivo assays that allow us to quantify Cryptosporidium, Toxoplasma, and Plasmodium microbial adhesion, microneme secretion, gliding motility, in vitro and in vivo infectivity, and to determine the mechanism by which the infectivity of these parasites is inhibited by L-PUFFA. Our results thus far provide hope that small molecule natural products, such as L-PUFFA or CDPK inhibitors will be valuable for the development of new drugs that show broad efficacy for treatment of apicomplexan parasitic diseases. Accordingly, L-PUFFA and CDPK inhibitors represent a unique opportunity to exploit a common mechanism used by the Apicomplexa to invade host cells (microneme secretion) and thus a potential "Achilles heel" target for the development of new anti-parasitic drugs. D. E. coli O157 Kansas Effects of a vaccine and a direct-fed microbial on fecal shedding of E. coli O157:H7 in pens of commercial feedlot cattle fed a diet supplemented with distillers grains. Our objective was to determine the efficacy of a siderophore receptor and porin (SRP®) proteins-based vaccine (VAC) and a direct-fed microbial (DFM; Bovamine®) for controlling fecal shedding of E. coli O157:H7 in pens of feedlot cattle fed a corn-based diet with 25% distillers grains. We conclude that the SRP vaccine can reduce fecal shedding of E. coli O157:H7 in potentially high risk populations of feedlot cattle. Fecal shedding of Escherichia coli O26 in feedlot cattle from a field trial evaluating an Escherichia coli O157:H7 vaccine and a direct-fed microbial. Preliminary analysis demonstrated no significant effects of treatment, or treatment by sampling week interaction, on O26 prevalence. However, a significant (P < 0.01) sampling week effect was observed with higher O26 prevalence in the two weeks prior to harvest compared to the previous two weeks. Preliminary results suggest the treatments had no significant effect on O26 shedding; however, these results are based on pooled colonies and ongoing analysis of individual isolates should provide further detail. Modeling the effect of bacterial transfer rates and interventions on the prevalence and concentration of Escherichia coli O157 on beef carcasses. We constructed a risk assessment model using @Risk in Microsoft Excel. Parameter distributions were developed from the existing scientific literature and incorporated into a Monte-Carlo framework. Our results indicate that the percentage of bacteria transferred between hides and carcasses is influential to carcass prevalence and concentration; however, there is little data to inform the distribution of this transfer. As expected, the sensitivity analysis showed successful carcass intervention has the highest impact on final carcass prevalence and concentration. Nebraska Single nucleotide polymorphism analysis for characterization of clinical versus bovine-biased genotypes. Two hundred thirty-one E. coli O157:H7 bovine isolates from a vaccine efficacy study (Moxley et al. 2009. Foodborne Pathog. Dis. 6:879-84) were submitted to the Washington station for single nucleotide polymorphism analysis to determine the proportions that may be classified as clinical versus bovine-biased genotypes. This collaboration is part of a USDA-AFRI-NIFA grant that was awarded to Washington State University and the University of Nebraska-Lincoln, with WSU as the lead institution and Dr. Tom Besser as Principal Investigator Efficacy testing of a proprietary compound for protection of gnotobiotic piglets against brain lesions caused by Shiga toxin-producing E. coli O157:H7 infection. Histopathological examinations on sections of formalin-fixed tissues from 20 of 45 gnotobiotic piglets were conducted with results provided to the South Dakota station. Piglets were randomly divided into principals and controls; principals received a proprietary compound for designed to protect against Shiga toxin-induced vascular damage. Controls received a placebo treatment. Dr. Moxley, station representative at Nebraska, served as pathologist, and was blinded to animal assignment to treatment and control groups. This collaboration is part of a grant awarded by the Teijin Corporation to South Dakota State University as lead institution with Dr. David Francis as Principal Investigator. E. Non-O157 STEC Nebraska Intimin type characterization of non-O157 STEC isolates. Non-O157 STEC are enzootic in cattle and constitute an emerging zoonotic threat. A current research goal of the USDA-NIFA is the development of pre-harvest interventions for STEC O26, O45, O103, O111, O121, O145, and O157:H7. Preliminary studies were conducted to characterize the STEC isolates from each of these seven serogroups for intimin type since this may influence the site and extent of intestinal colonization, and thereby constitute a suitable target for vaccines or other intervention strategies. F. Norovirus Ohio The effects of simvastatin or interferon-± on infectivity of human norovirus in the gnotobiotic pig model for the study of antivirals. Our study demonstrated that use of simvastatin enhanced HuNoV infectivity in the Gn pig model. Thus, its use may also support growth of HuNoV in cell culture. We also verified that the increased infectivity of HuNoV might be associated with the inhibitory effect of statins on innate immunity (IFN-±). This observation might explain the exacerbated HuNoV disease and the related higher mortality in statin-treated humans. In addition, we showed that oral treatment with rhIFN-± can curtail early HuNoV fecal shedding in the Gn pig model. This approach could be applicable to control multiple genogroups and genotypes of HuNoVs, including the GII.4 variants that have emerged each year. Collectively, HuNoV infectivity in Gn pigs can be enhanced by simvastatin treatment or reduced by oral treatment with rhIFN-±. These observations indicate that Gn pigs are a useful model to test the efficacy of antivirals against HuNoV. H. L. intracellularis Minnesota Infection of sparrows (Passer domesticus) and different mice strains with L. intracellularis. The susceptibility of sparrows (Passer domesticus) and four strains of mice (Swiss, BALB/c, C-57 and DB-A) to L. intracellularis infection were studied. We found that sparrows do not seem to be relevant in the epidemiology of L. intracellularis. The results showed variations in the establishment of lesions among the four different mice strains used. Transcriptional profiling of a pathogenic and an attenuated homologous L.intracellularis isolate during in vitro infection. The current study used high-throughput sequencing technology to characterize the transcriptional profiling of a pathogenic and an attenuated isolate during in vitro infection. We identified distinct genes and pathways between a pathogenic and an attenuated L. intracellularis isolate. This information supports our hypothesis and opens a new research field for studying target genes involved in the ecology, pathogenesis and physiology of L. intracellularis. Evidence of host adaptation in L. intracellularis infections. The objective of this study was to evaluate the susceptibilities of pigs and horses to L. intracellularis infection using porcine and equine isolates. In the pig trial, all animals infected with the porcine isolate demonstrated watery diarrhea and seroconversion from 21 days PI. No clinical signs or seroconversion were observed in equine isolate-infected pigs. Two pigs infected with porcine isolate were euthanized on day 21 PI and showed intestinal proliferative lesions associated with the presence of Lawsonia-specific antigen in the intestinal epithelium identified by IHC. No macroscopic or histologic lesions, including IHC staining, were observed in two equine isolate-infected pigs which were also euthanized 21 days PI. Marked clinical signs, longer periods of bacterial shedding and stronger serologic immune responses were observed in animals infected with species-specific isolates. Intracellularis increases Salmonella enterica levels in the intestines of pigs. To determine if there was an association between S. enterica carriage promoted by infection with L. intracellularis, we measured S. enterica loads in 4 intestinal sites of pigs challenged with L. intracellularis (at 5 weeks of age), S. enterica serovar Typhimurium (at 6 weeks of age), or both microbes. Preliminary data demonstrated multiple changes in the intestines of pigs challenged with one or both pathogens compared to non-challenged controls. As well, there were differences in the microbiome compositions between the challenge groups. I. Rotavirus Kansas Inhibitory effects of bile acids and synthetic Farnesoid X receptor agonists on rotavirus replication. Rotaviruses (group A rotaviruses) are the most important cause of severe gastroenteritis in infants and children worldwide. We examined the effects of bile acids and synthetic FXR agonists on rotavirus replication in association with cellular lipid levels. In a mouse model of rotavirus infection, effects of oral administration of CDCA on fecal rotavirus shedding were investigated. We conclude that bile acids and FXR agonists play important roles in the suppression of rotavirus replication, which may serve therapeutic option for rotavirus infection in animals and humans. Ohio Probiotics: Effect on neonatal gut immunity to RV. Prior trials have shown satisfactory protection rates for live oral RV vaccines in developed countries. However, the cost is high and the effectiveness is low for RV vaccines tested in underdeveloped countries where most needed. Thus, alternative low cost treatments to moderate HRV diarrhea or improve vaccine efficacy are urgently needed. An improved understanding of the influence of probiotic bacteria on HRV vaccines and disease in neonatal Gn pigs will permit an extrapolation to interventions for infants to prevent HRV diarrhea. Rotavirus Vaccine Efficacy in Neonates: Impact of Vitamin A. Our results suggest that maternal and childhood VAD may decrease HRV vaccine efficacy and exacerbate HRV diarrheal disease. Vitamin A deficiency in children from developing countries is high (11-40%) and may result in reduced efficacy of HRV vaccines and increased severity of HRV disease. A similar scenario is possible in farm animals, especially multiparous animals, but has not been addressed. The association between RV related diarrheal disease and VAD in swine and cattle also should be assessed and vitamin A supplementation may be implemented as a vaccine adjuvant and an intervention strategy. Objective 3. Focus on disseminating knowledge  Provide training or continuing education opportunities and dissemination of information to students, producers, veterinarians, and diagnostic laboratories Michigan Several students were mentored for study of enteric diseases of food animals. Jamie Jennifer Kopper (DVM candidate) successfully defended her PhD thesis and will go on to finish two more years of veterinary school. Four other students are pursuing PhD degrees in food safety related to these enteric pathogens including John Paul Jerome, Jessica St. Charles, Ankit Malik and Barbie Gadsden, DVM. Three of these students have passed their preliminary exams, two during 2011. One is preparing for this exam. Four undergraduate students are working in the lab on these projects. One undergraduate was accepted to and started medical school during 2011. Dr. Mansfield organized a Spring seminar in Food and Waterborne Diseases for the faculty and students of Michigan State University. People attending have come from the Agricultural, Veterinary Medicine, Human Medicine, Microbiology and Food Science, and Human Nutrition departments. The speakers included Christopher Waters, PhD (Biofilms in foodborne bacteria) and John Paul Jerome (Contingency genes in Campylobacter jejuni). Also, Dr. Mansfield attended and presented at a scientific conference held by the National Institutes of Health in Baltimore, Maryland on Enteric Diseases. Dr. Mansfield helped to organize and attended the USDA Rushmore Enteric Diseases Meeting NC1041 in Chicago, Illinois on December 3rd and 4th. Her graduate student John Paul Jerome gave a talk about his work entitled Contingency loci dynamics during Campylobacter jejuni infection. Dr. Mansfield gave a talk for the Department of Large Animal Clinical Sciences at Michigan State University entitled Investigating the role of Campylobacter jejuni in acute and chronic disease: the genetic basis of pathotypes on 02/01/2011. North Dakota The course International Animal Production, Disease Surveillance and Public Health was offered for the fifth time in summer 2011 with 9 US students. The joint Master of Science (M.S.) degree as well as a graduate certificate in International Infectious Disease Management and Biosecurity (2008-2011) program was approved by the ND State Board of Higher Education, The first group of students (8) enrolled in fall, 2011. 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. 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 Conference of Research Workers in Animal Disease, 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. They have disseminated new information, reagents, and procedures to producers, industries, veterinary diagnostic laboratories and veterinarians (both swine and equine).

Wilson DL, Rathinam VK, Qi W, Wick LM, Landgraf J, Bell JA, Plovanich-Jones A, Parrish J, Finley RL, Mansfield LS and Linz JE. 2010. Genetic diversity in Campylobacter jejuni is associated with differential colonization of broiler chickens and C57BL/6J IL-10 deficient mice. Microbiology. 2010 Jul;156(Pt 7):2046-57. Jerome, J.P., J.A. Bell, A.E. Plovanich-Jones, J.E. Barrick, C.T. Brown, L.S. Mansfield. 2010. Standing genetic variation in contingency genes drives the adaptation of Campylobacter jejuni to a novel host. PLoS ONE 6(1): e16399. doi:10.1371/journal.pone.0016399. Bell, J.A., J. R. Gettings, J. P. Jerome, J. J. Kopper, A. Plovanich-Jones, V. A. K. Rathinam, J. L. St. Charles, E.J. Smith, A. G. Staunton, and Mansfield, L.S. Outcome of Campylobacter jejuni infection of C57BL/6 IL-10-/- mice varies with strain. In review, Infection and Immunity. Dodd C, Sanderson MW, Jacob M, Renter DG. Modeling preharvest and harvest interventions for Escherichia coli O157 contamination on beef cattle carcasses. J Food Prot. 2011; 74(9):1422-1433. Nickell JS, White BJ, Larson RL, Renter DG, Sanderson MS, Peck C. Bovine Viral Diarrhea Virus (BVDV) Status in cow-calf herds and associations with biosecurity and production practices among Montana beef producers. Bov Pract. 2011; 45(1): 14-22. Dodd C, Renter DG, Shi X, Alam MJ, Nagaraja TG, Sanderson MW. Prevalence and persistence of Salmonella in cohorts of feedlot cattle. Foodborne Pathog Dis. 2011; 8(7): 781-789. Nickell JS, White BJ, Larson RL, Renter DG, Sanderson MW. A simulation model to quantify the value of implementing whole-herd Bovine Viral Diarrhea Virus testing strategies in beef cowcalf herds. J Vet Diagn Invest. 2011; 23(2): 194-205. Dodd CC, Renter DG, et al. Evaluation of the effects of a commercially available Salmonella Newport siderophore receptor and porin protein vaccine on fecal shedding of Salmonella bacteria and health and performance of feedlot cattle. Am J Vet Res., 2011; 72(2): 239-247. Taylor E, Renter DG, Nagaraja TG. Genetic variations in shiga toxin-producing abilities of bovine and human Escherichia coli O157:H7. Zoonoses Public Health, 2011; 58(3): 185-91. Sargeant JM, OConnor A, Renter DG, Kelton D, Snedeker K, Wisener L, Leonard E, Guthrie A, Faires M. Reporting of methodological features in observational studies of pre-harvest food safety. Prev Vet Med., 2011; 98(2-3): 88-98. Paddock, Z. D., C. E. Walker, J. S. Drouillard, Nagaraja T.G.. Dietary monensin level, supplemental urea, and ractopamine on fecal shedding of Escherichia coli O157:H7 in feedlot cattle. J. Anim. Sci., 2011; 89:2829-2835. Amachawadi, R. G., N. W. Shelton, X. Shi, J. Vinasco, S. S. Dritz, M. D. Tokach, J. L. Nelssen, H. M. Scott, T. G. Nagaraja. Selection of tcrB gene mediated copper resistant fecal enterococci in pigs fed diets supplemented with copper. Appl. Environ. Microbiol., 2011; 77:5597-5603. Jacob, M. E., K. M. Almes, X. Shi, J. M. Sargeant, and T. G. Nagaraja. Escherichia coli O157 diversity in bovine fecal samples. J. Food Prot., 2011; 74:1186-1188. Yunjeong Kim and Kyeong-Ok Chang. Inhibition of Rotavirus replication by Bile Acids through Farnesoid X Recepter. J Virol., 2011; 85(23):12570-7. Aqeel Ahmad, Anuradha Ghosh, Coby Schal, and Ludek Zurek. Insects in confined swine operations carry a large antibiotic resistant and potentially virulent enterococcal community. BMC Microbiol., 2011; 11(1):23. Daisuke Takahashi, Yunjeong Kim, Kyeong-Ok Chang, Asokan Anbanandam, Om Prakash. 2011. 1H, 15N, and 13C resonance assignments of Norwalk virus protease. Biomol NMR Assign. Jun 8. [Epub ahead of print] Jacob, M. E., X. Shi, B. An, T. G. Nagaraja, and J. Bai. Evaluation of a multiplex real-time PCR for the quantification of Escherichia coli O157 in cattle feces. Foodborne Path. Dis. In Press. Paddock, Z., X. Shi, J. Bai, and T.G. Nagaraja. Applicability of a multiplex PCR to detect O26, O45, O103, O111, O121, O145, and O157 serogroups of Escherichia coli in cattle feces. Vet. Microbiol. In Press. Kyeong-Ok Chang, Daisuke Takahashi, Om Prakash, and Yunjeong Kim. Characterization of Proteases from Norovirus Genogroup I and II with the Fluorescence Resonance Energy Transfer Assay. Virology. In Press. Vannucci FA, Wattanaphansak S, Gebhart CJ. 2012. An alternative method for cultivation of Lawsonia intracellularis. J. Clin. Microbiol. (accepted). Paradis M, Gebhart CJ, Toole D, Vessie G, Winkelman NL, Bauer SA, Wilson JF, McClure CA. 2011. Subclinical Ileitis: Diagnostic and performance parameters in a multi-dose mucosal homogenate challenge model. (accepted). Pusterla N, Mapes S, Gebhart C. 2011. Lawsonia intracellularis-specific interferon c gene expression by peripheral blood mononuclear cells in vaccinated and naturally infected foals. Vet. J. (in press). Nogradi N, Slovis NM, Gebhart CJ, Wolfsdorf KE, McCrcken JL, Scoggin CF, Kass PH, Mapes SM, Toth B, Lundquist ML, Pusterla N. 2011. Evaluation of the field efficacy of an avirulent live Lawsonia intracellularis vaccine in foals. Vet. J. (in press). Pusterla N, Vannucci FA, Mapes SM, Nogradi N, Collier J, Hill J, DiFrancesco M, White A, Akana N, Simonek G, Gebhart C. 2011. Evaluation of an avirulent live vaccine of Lawsonia intracellularis in the prevention of equine proliferative enteropathy in experimentally infected weanling foals. A. J. Vet. Res. (in press). Page AE, Stillst HF, Chander Y, Gebhart CJ, Horohov DW. 2011. Adaptation and validation of a bacteria-specific enzyme-linked immunosorbent assay for determination of farm-specific Lawsonia intracellularis seroprevalence in central Kentucky Thoroughbreds. Equine Vet. J. 43 Suppl 40: 25-31. McOrist S, Blunt R, Gebhart CJ. 2011. Pig-associated Lawsonia intracellularis in various on-farm dipterous fly stages. J. Swine Health Prod. 19:277-283. Page AE, Loynachan AT, Bryant U, Stills HF Jr, Adams AA, Gebhart CJ, Pusterla N, Horohov DW. 2011. Characterization of the interferon gamma response to Lawsonia intracellularis using an equine proliferative enteropathy challenge (EPE) model. Vet. Immunol. Immunopathol. 143:55-65. Page AE, Slovis NM, Gebhart CJ, Wolfsdorf K, Mapes SM, Pusterla N. 2011. Serial use of serologic assays and fecal PCR assays to aid in identification of subclinical Lawsonia intracellularis infection for targeted treatment of Thoroughbred foals and weanlings. J. Am. Vet. Med. Assoc. 238:1482-9. Esseili, M. A., Q. Wang, and L. J. Saif. 2011. Human GII.4 norovirus-like-particles (VLPs) bind to carbohydrates of Romaine lettuce leaf cell wall materials. Appl Environ Microbiol. (Accepted) 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. Zhang, W., Azevedo, M.S., Wen, K., Gonzalez, A., Saif, L.J., Li, G., Yousef, A.E., Yuan, L. 2008a. Probiotic Lactobacillus acidophilus enhances the immunogenicity of an oral rotavirus vaccine in gnotobiotic pigs. Vaccine. 2008 Jul 4;26(29-30):3655-61. Zhang, W., Azevedo, M.S., Gonzalez, A.M., Saif, L.J., Van Nguyen, T., Wen, K., Yousef, A.E., Yuan, L. 2008b. Influence of probiotic Lactobacilli colonization on neonatal B cell responses in a gnotobiotic pig model of human rotavirus infection and disease. Vet Immunol Immunopathol. Mar 15;122(1-2):175-81. Zhang, W., Wen, K., Azevedo, MS., Gonzalez, A., Saif, L.J., Li, G., Yousef, A.E., Yuan, L. 2008c. Lactic acid bacterial colonization and human rotavirus infection influence distribution and frequencies of monocytes/macrophages and dendritic cells in neonatal gnotobiotic pigs. Vet Immunol Immunopathol. 2008 Feb 15;121(3-4):222-31. Yasser M Sanad, ssmat I Kassem, Melanie Abley, Wondwossen Gebreyes, Jeffrey T. LeJeune, Gireesh Rajashekara. 2011. Genotypic and Phenotypic Properties of Beef Cattle-Associated Campylobacter and their Implications to Public Health in the USA. PLoS One: 6(10):e25778. Epub 2011 Oct 19. Mary Drozd, Dharanesh Gangaiah, Zhe Liu, and Gireesh Rajashekara. 2011. Contribution of PhoX to Twin Arginine Translocation mediated Campylobacter jejuni function and resilience to Environmental Stresses. PLoS One;6(10):e26336. Epub 2011 Oct 20. Yasser Sanad, Issmat I. Kassem, Jun Lin, Jeffrey T. LeJeune, Gireesh Rajashekara. 2011. Occurrence of the invasion associated marker (iam) in Campylobacter jejuni isolated from cattle. BMC Res Notes (In Press). Issmat I. Kassem, Yasser Sanad, Dharanesh Gangaiah, Michael Lilburn, Jeffery Lejeune, Gireesh Rajashekara. 2010. Use of bioluminescence imaging to monitor Campylobacter survival in chicken litter. J Appl Microbiol. 109(6):1988-97. Butler, JE, K Santiago-Mateo, X-Z Sun, N Wertz, M Sinkora, and DH Francis. 2011. Antibody repertoire development in fetal and neonatal piglets. XX. B cell lymphogenesis is absent in the ileal Peyers Patches, their repertoire development is antigen dependent, and they are not required for B cell maintenance. J Immunol 187:5141-5149. Butler JE, X Sun, N Wertz, KM Lager, K Chaloner, J Urban Jr, DH Francis, PL Nara, GJ Tobin. 2011. Antibody repertoire development in fetal and neonatal piglets XXI. Usage of most VH genes remains constant during fetal and postnatal Development. Molecular Immunology 49: 483-494. Mateo KS, JH Ayres, M Zhao, JE Butler and DH Francis. 2011. Intestinal resection and anastomosis in neonatal gnotobitic piglets. J Am Assoc Lab An Sci. 50: 361-354. Cooper, K.K., Cooper, M.A., Zuccolo, A., Law, B., L.A. Joens. Complete Genome Sequence of Campylobacter jejuni Strain S3. J Bacteriol. 193(6):1491-2. 2011. Brillhart, C.D., L.A. Joens. Prevalence and characterization of Salmonella serovars isolated from oysters served raw in restaurants. J Food Prot. 74(6):1025-9. 2011. Morrison, C.M., Armstrong, A.E., Evans, S., Mild, R.M., Langdon, C.J., L.A. Joens. Survival of Salmonella Newport in oysters. Int J Food Microbiol. Epub May 13. 2011. Mild, R.M., L.A. Joens, Friedman, M., Olsen, C.W., McHugh, T.H., Law, B., Ravishankar S. Antimicrobial edible apple films inactivate antibiotic resistant and susceptible Campylobacter jejuni strains on chicken breast. J Food Sci. 76(3):M163-8. 2011. Theoret J.R., Cooper K.K., Glock RD, L.A. Joens. A Campylobacter jejuni Dps Homolog Has a Role in Intracellular Survival and in the Development of Campylobacterosis in Neonate Piglets. Foodborne Pathog Dis. 8(12): 1263-1268. 2011. Liu Y, Kuhlenschmidt, MS, Kuhlenschmidt, TB, and Nguyen, TH. (2011) Characterization of Cryptosporidium parvum Oocyst Wall Macromolecules and Adhesion Kinetics of Oocysts on Quartz Surface. Biomacromolecules. 11:2109-15. Bergner DW, Kuhlenschmidt TB, Firkins LD, and Kuhlenschmidt MS, (2011) Synthesis and characterization of a neoglycolipid that blocks porcine group A rotavirus infectivity Nutrients 3:228-244. Kowalewski MM, Salzer JS, Deutsch JC, M Raño,. Kuhlenschmidt MS, and Gillespie, TR. (2011) Black and gold howler monkeys (Alouatta caraya) as sentinels of ecosystem health: patterns of zoonotic protozoa infection relative to degree of humanprimate contact. Am. J. Primatology 73:75-83. Fuller KL, Kuhlenschmidt TB, Kuhlenschmidt MS, Jimenez-Flores R, and Donovan SM. (2011) Milk Fat Globule Membrane Isolated from Buttermilk or Cheese Whey and their Lipid Component Inhibit Infectivity of Rotavirus In Vitro. Nutrients (in press). Perez VG, Jacobs CM, Barnes J, Jenkins, Kuhlenschmidt MS, Fahey GC, and Pettigrew J (2011) Effect of corn distillers dried grains with solubles MC and Eimeria acervulina infection on growth performance and the intestinal microbiota of young chicks. J. Poultry Sci. 9:959-964. Bhattrai R, Kalita P, Trask J, Kuhlenschmidt MS (2011) Development of a physically-based model for transport of Cryptosporidium parvum in overland flow. Environ Modell SoftW 26:1289-1287. Erume, J., P. Wijemanne, E. M. Berberov, S. D. Kachman, D. J. Oestmann, D. H. Francis, and R. A. Moxley. 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. (submitted) Moxley, R. A. Chapter 7, Family Enterobacteriaceae, Veterinary Microbiology, 3rd Edition, Wiley-Blackwell Publishing, John Wiley & Sons, Hoboken, NJ. D. S. McVey, M. Kennedy, and M. M. Chengappa (Eds). (submitted) Moxley, R. A. Chapter 8, Enterobacteriaceae: Escherichia, Veterinary Microbiology, 3rd Edition, Wiley-Blackwell Publishing, John Wiley & Sons, Hoboken, NJ. D. S. McVey, M. Kennedy, and M. M. Chengappa (Eds). (submitted) Moxley, R. A., Chapter 9, Enterobacteriaceae: Salmonella, 3rd Edition, Wiley-Blackwell Publishing, John Wiley & Sons, Hoboken, NJ. D. S. McVey, M. Kennedy, and M. M. Chengappa (Eds). (submitted) Moxley, R. A., Chapter 10, Enterobacteriaceae: Yersinia, 3rd Edition, Wiley-Blackwell Publishing, John Wiley & Sons, Hoboken, NJ. D. S. McVey, M. Kennedy, and M. M. Chengappa (Eds). (submitted) Moxley, R. A., Chapter 11, Enterobacteriaceae: Shigella, 3rd Edition, Wiley-Blackwell Publishing, John Wiley & Sons, Hoboken, NJ. D. S. McVey, M. Kennedy, and M. M. Chengappa (Eds). (submitted) Hoang, K.V., N. J. Stern, and J. Lin. 2011. Development and stability of bacteriocin resistance in Campylobacter. Journal of Applied Microbiology.111:1544-1550. Lin, J. 2011. Effect of antibiotic growth promoters on intestinal microbiota in food animals: a novel model for studying the relationship between gut microbiota and human obesity? Frontiers in Cellular and Infection Microbiology. Vol. 2 Article 53. Hoang, K.V., N. J. Stern, A. M. Saxton, F. Xu, X. Zeng, and J. Lin. 2011. Prevalence, development, and molecular mechanisms of bacteriocin resistance in Campylobacter. Applied and Environmental Microbiology 77:2309-2316. Burrough, E.R., K. DiVerde, O. Sahin, P. J. Plummer, Q. Zhang, and M. J. Yaeger, DVM. 2011. Expression of Toll-like receptors 2 and 4 in subplacental trophoblasts from guinea pigs aborting following infection with Campylobacter jejuni. Vet. Pathol. 48(2):381-8 Jeon, Byeonghwa, Yang Wang, Haihong Hao, Yi-Wen Barton, Qijing Zhang. 2011. Contribution of CmeG to antibiotic and oxidative stress resistance in Campylobacter jejuni. J. Antimicrob. Chemother. 66 (1):79-85. Burrough, Eric R., Orhan Sahin, Paul J. Plummer, Kevin DiVerde, Qijing Zhang, Michael J. Yaeger. 2011. Comparison of two commercial ovine Campylobacter vaccines and an experimental bacterin in a guinea pig model. Am. J. Vet. Res. 72: 799-805. Plummer, Paul, Jinge Zhu, Masato Akiba, Dehua Pei, and Q. Zhang. 2011. Identification of a Key Amino Acid of LuxS Involved in AI-2 Production in Campylobacter jejuni. PLoS ONE 6 (1): e15876. doi:10.1371/journal.pone.0015876. Muraoka Wayne T. and Qijing Zhang. 2011. Phenotypic and genotypic evidence for L-fucose utilization by Campylobacter jejuni. J. Bacteriol. 193(5):1065-75. Qin, Shang-Shang, Cong-Ming Wua, Yang Wang, Byeonghwa Jeon, Zhang-Qi Shen, Yu Wang, Qijing Zhang, Jian-Zhong Shen. 2011. Antimicrobial resistance in Campylobacter coli isolated from pigs in two northern provinces of China. International Journal of Food Microbiology. 146: 94-98. Hsiang-Ting Lei, Zhangqi Shen, Priyanka Surana, Mathew D. Routh, Chih-Chia Su, Qijing Zhang and Edward W. Yu. 2011. Crystal structures of CmeR-bile acid complexes from Campylobacter jejuni. Protein Science 20: 712-23. Oakland M, B. Jeon, O. Sahin, Z. Shen, and Q. Zhang. 2011. Functional Characterization of a Lipoprotein-Encoding Operon in Campylobacter jejuni. PLoS One. 6(5): e20084. doi:10.1371/journal.pone.0020084. Shen, Zhangqi, Xiaoying Pu, and Qijing Zhang. 2011. Salicylate functions as an efflux pump inducer and promotes the emergence of fluoroquinolone-resistant mutants in Campylobacter jejuni. Appl. Environ. Microbiol. 77:7128-7133. Kassem, Issmat I, Qijing Zhang & Gireesh Rajashekara. 2011. The twin-arginine translocation system: contributions to the pathobiology of Campylobacter jejuni. Future Microbiology 11: 1315-1327. Burrough, Eric R, Zuowei Wu, Orhan Sahin, Qijing Zhang, and Michael J. Yaeger. 2011. 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 Campylobacter jejuni. Vet Pathol 2011; published November 11 as doi:10.1177/0300985811424755.