Source: OHIO STATE UNIVERSITY submitted to
EMERGING AND RE-EMERGING ZOONOTIC DISEASES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
ANIMAL HEALTH
Reporting Frequency
Annual
Accession No.
0196668
Grant No.
(N/A)
Project No.
OHO00003-AH
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2008
Project End Date
Sep 30, 2013
Grant Year
(N/A)
Project Director
Rajashekara, GI.
Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691
Performing Department
Food Animal Health Research Program
Non Technical Summary
Diseases transmitted through food borne/waterborne routes, constitute a significant burden for public health. Over two-thirds of all infectious agents affecting humans have animal reservoir. Importantly, treatment of diseases caused food/waterborne microbes is complicated either by the emergence of resistant bacteria or lack of effective treatments available for viruses. These public health and safety concerns underscore the critical need for integrated research efforts to better understand the ecology, epidemiology and pathogenesis of disease that are communicable to humans through food animals and water, so that effective strategies can be developed to prevent the dissemination of these organisms in the environment and limit the spread of these agents to humans. The over all goal of this project is to understand the epidemiology, ecology and pathogenesis of medically important infectious agents in animal reservoirs and the environment including; investigation of the occurrence, dissemination and ecological fitness of antibiotic resistant pathogens; and design of efficient methods/strategies and models for the detection and prevention of pathogens that are communicable to humans.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7124030110120%
7124030116020%
7124030117020%
7223599110010%
7223399110010%
7223299110110%
7223299110010%
Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins; 722 - Zoonotic Diseases and Parasites Affecting Humans;

Subject Of Investigation
3299 - Poultry, general/other; 4030 - Viruses; 3599 - Swine, general/other; 3399 - Beef cattle, general/other;

Field Of Science
1101 - Virology; 1160 - Pathology; 1100 - Bacteriology; 1170 - Epidemiology;
Goals / Objectives
The primary goal of this research is to exploit the research expertise of the Food Animal Health Research Program to enhance the understanding of the microbiology, epidemiology and pathogenesis and control of infectious agents of livestock that have potential for zoonotic transmission, including several pathogens that may be used as agents of agribioterrorism. To achieve this goal we propose to investigate the following aspects of zoonotic agents harbored by food-producing animals: 1. Mechanisms of microbial pathogenesis of zoonotic agents of food safety concerns including the production of bacterial toxins 2. Immune response using animal models of infection and immunity 3. Epidemiology and natural history of infection and interspecies transmission of zoonotic pathogens 4. Development of vaccines, therapeutics, adjuvants, and diagnostics for food safety and agribiodefense.
Project Methods
Clearly, due to the wide variety of zoonotic pathogens that may emerge as potential public health threats, the specific methods will vary from individual project to project. However, our underlying strategy is to use a hypothesis-driven research approach to provide sound scientific data concerning the research problems investigated. Research may include development and/or utilization of animal models of infection to characterize mechanisms of pathogenesis and pathogen/host interactions. These include studies of pathogenesis and comparative pathogenesis, interspecies transmission of pathogens, understanding the molecular basis of pathogenesis, studies of pathogen and host gene expression and characterization of host genetic risk factors for disease susceptibility. We will exploit current advances in the detection of gene and protein expression patterns to assess disease progression, and to seek unique host cellular signatures indicative of infection and/or protective immune responses. Studies of the natural rates of horizontal gene transfer, especially of plasmid-encoded toxins and resistance factors, in the microbial ecosystems likely to be most exposed to agents of agribioterrorism For the studies of natural history of microbial pathogens and epidemiology of infections in natural non-human and human hosts, we will use extensive in-field sampling strategies coupled with epidemiological practices targeted at identifying factors that influence the distribution, acquisition, course and severity of human disease. This will also include studies on pathogens in the environment and non-food-producing vectors, when those studies may contribute to the understanding of factors bearing on emergence, transmission, persistence, development of virulence, host switching, and/or epidemic/epizootiologic thresholds as they directly pertain to human disease.

Progress 10/01/08 to 09/30/13

Outputs
Target Audience: Livestock Producers and processors, Scientists, Veterinarians, Veterinary Diagnosticians, bacterialogists, virologists, Immunologists, Exrtension officers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Graduate and undergraduate students and postdoctoral fellows acquired research skills in the area of epidemiology, diagnostics, vaccine development, pre- and post harvest control strategies, as well as in basic research relating to mechanisms of disease pathogenesis. These trainings resulted in the completion of several Masters and Graduate students dissertation and endured them with skills to conduct independent research. Similarly researchers were trained in extension and education such that they can effectively communicate with different stakeholders. Several invention disclosures were filed during this period and some were awarded with patent for the novelty and its significance either to livestock industry or public health. y, zoonotic diseases, bacterial pathogenesis, genomics, and emerging infectious diseases. Several investigators contributed to the editorship and/or Ad Hoc reviews of peer reviewed journal articles, white papers, and fact sheets Products: Number of novel findings was reported that addresses both the basic as well as applied needs of zoonotic and eme How have the results been disseminated to communities of interest? Several meetings were conducted with the stakeholders including livestock and poultry producers as well as veterinarians. Written and electronic media was distributed to stakeholders and research findings were published in peer reviewed journals. The knowledge gained and communicated to different stakeholders is expected to enhance food safety, reduce risk of zoonoses, aid in the development of vaccines and therapeutic against human infections and preparedness against emerging and re-emerging infectious disease of livestock as well as public health significance. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We employed interdisciplinary research approach to address food safety concerns, particularly: 1) developed and validated a toolbox of assays that permit the rapid detection, quantification and characterization of pathogens in food and water, 2) used a multifaceted approach to tackle the problem of antibiotic resistance; for example, (a) use of a high throughput method to screen thousands of naturally small molecules for their effectiveness to control important foodborne pathogens such Salmonella and Campylobacter, and (b) determined the role of wildlife in transferring antibiotic resistant bacteria to cattle and pigs to develop methods to prevent spread of drug-resistant bacterial pathogens in food animals, 3) used piglet model to provide insight on the important role that probiotic bacteria play in the protection from infectious diseases, 4) understand the extremely complex interactions between the microorganisms present in the gut, the genetic make-up of the host, and the host resistance or susceptibility to infectious diseases, 5) interdisciplinary approach involving social and biological sciences to enhance adoption of produce safety behaviors from farm to table, 6) developed a biodegradable biocompatible nanoparticle-based killed PRRSV (NanoPRRS) vaccine, 7) standardized and validated a PRRSV neutralizing antibody detection assay in pen-based pig oral fluid samples, which could be applied to monitor PRRS herd immune status in swine herds including in litters, 8) generated M2e-based P particle (M2e-P) vaccines of different M2e consensus sequence and tested in animal models for a broad protection against a wide range of avian, swine and human flu viruses, 9) Defined the pathogenesis of GIII.2 bovine norovirus, CV186-OH/00/US strain using gnotobiotic calves, 10) Developed methods for detection of porcine epidemic diarrhea virus (PEDV) in US swine, and developed gnotobiotic pig model to understand its pathogenesis and 11) Studies molecular epidemiology of Campylobacter from various food producing animals such as beef and dairy cattle, chickens and turkeys.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Kassem II, Chandrashekhar K, Rajashekara G. Of energy and survival incognito: a relationship between viable but non-culturable cells formation and inorganic polyphosphate and formate metabolism in Campylobacter jejuni. Front Microbiol. 2013 Jul 9;4:183.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Chattha KS, Vlasova AN, Kandasamy S, Rajashekara G, Saif LJ. 2013. Divergent immunomodulating effects of probiotics on T cell responses to oral attenuated human rotavirus vaccine and virulent human rotavirus infection in a neonatal gnotobiotic piglet disease model. J Immunol. 2013 Sep 1;191(5):2446-56.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Chattha KS, Vlasova AN, Kandasamy S, Esseili MA, Siegismund C, Rajashekara G, Saif LJ.. 2013. Probiotics and colostrum/milk differentially affect neonatal B cell responses to oral rotavirus vaccine. Vaccine. 2013 Apr 8;31(15):1916-23.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Petersen E, Rajashekara G, Sanakkayala N, Eskra L, Harms J, Splitter G.. 2013. Erythritol triggers expression of virulence traits in Brucella melitensis. Microbes and Infection. Jun;15(6-7):440-9.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Annamalai T, Pina-Mimbela R, Kumar A, Binjawadagi B, Liu Z, Renukaradhya GJ, Rajashekara G. 2013. Evaluation of nanoparticle encapsulated OMPs for the control of Campylobacter jejuni colonization in chickens. Poult Sci. Aug;92 (8):2201-11.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Homan HJ, LeJeune JT, Pearl DL, Seamans TW, Slowik AA, Morasch MR, Linz GM. 2013. Use of dairies by postreproductive flocks of European starlings. J Dairy Sci. ;96(7):4487-93.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Oka T, Saif LJ, Wang Q. 2013. First Complete Genome Sequence of a Genogroup II Genotype 18 Porcine Norovirus, Strain QW125. Genome Announc. 2013 Jun 13;1(3).
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Scheuer KA, Oka T, Hoet AE, Gebreyes WA, Molla BZ, Saif LJ, Wang Q. 2013. Prevalence of porcine noroviruses, molecular characterization of emerging porcine sapoviruses from finisher swine in the United States, and unified classification scheme for sapoviruses. J Clin Microbiol.;51(7):2344-53.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Vlasova AN, Chattha KS, Kandasamy S, Siegismund CS, Saif LJ. 2013. Prenatally acquired vitamin A deficiency alters innate immune responses to human rotavirus in a gnotobiotic pig model. J Immunol. ;190(9):4742-53.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Prevalence and molecular characterization of porcine enteric caliciviruses and first detection of porcine kobuviruses in US swine. Arch Virol. 2013 Jul;158(7):1583-8.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: 19. Amimo JO, Vlasova AN, Saif LJ. 2013. Prevalence and genetic heterogeneity of porcine group C rotaviruses in nursing and weaned piglets in Ohio, USA and identification of a potential new VP4 genotype. Vet Microbiol. 2013 May 31;164(1-2):27-38.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Williams ML, Pearl DL, Bishop KE, Lejeune JT. 2013. Use of Multiple-Locus Variable-Number Tandem Repeat Analysis to Evaluate Escherichia coli O157 Subtype Distribution and Transmission Dynamics Following Natural Exposure on a Closed Beef Feedlot Facility. Foodborne Pathog Dis. Jul 16. 2013
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Won G, Schlegel PJ, Schrock JM, LeJeune JT. 2013. Absence of direct association between coliforms and Escherichia coli in irrigation water and on produce. J Food Prot: 76(6):959-66.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: 24. Ali A, Ibrahim M, Eladl AE, Saif YM, Lee CW. 2013. Enhanced replication of swine influenza viruses in dexamethasone-treated juvenile and layer turkeys. Vet Microbiol. 2013 Mar 23;162(2-4):353-9.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Manickam C, V. Dwivedi, J. Miller, T. Papenfuss and G.J. Renukaradhya (2013) Mycobacterium tuberculosis whole cell lysate enhances proliferation of CD8 positive lymphocytes and nitric oxide secretion in the lungs of live porcine respiratory and reproductive syndrome virus vaccinated pigs. Viral Immunol., 26(1):102-108.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Renukaradhya G.J, M.A. Khan, R.M. Gallo, D. Shaji, J. Liu, R.R. Brutkiewicz (2013). Forming a complex with MHC class I molecules interferes with mouse CD1d functional expression. PLoS One. 8(8):e72867.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Olsen C, L. Karriker, C. Wang, G.J. Renukaradhya, B. Binjawadagi, A. Kittawornrat, S. Lizano, J. Coetzee, R. Main, A. Meiszberg, Y. Panyasing, J. Zimmerman (2013). Effect of collection material on pig oral fluid testing results. Veterinary Journal, 198(1):158-163.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Drozd M, Merrick NN, Sanad YM, Dick LK, Dick WA, Rajashekara G. Evaluating the occurrence of host-specific , general fecal indicators, and bacterial pathogens in a mixed-use watershed. J Environ Qual. 2013 May-Jun;42(3):713-25.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Sanad YM, Closs G Jr, Kumar A, LeJeune JT, Rajashekara G. Molecular epidemiology and public health relevance of Campylobacter isolated from dairy cattle and European starlings in Ohio, USA. Foodborne Pathog Dis. 2013 Mar;10(3):229-36.


Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: Activities: A large number of hypothesis-driven experiments that are both basic as well applied in nature were conducted during this reporting period. As a result, we published several peer reviewed journal articles as well as findings were presented at the local, regional, national and international meetings. During this period a large number of Postdoctoral fellows, Graduate students, Undergraduate students and High school students were mentored in various projects in areas of emerging and re-emerging zoonotic diseases. Events: Presentation of research findings to various stakeholders (livestock producers, food processors and policy makers) and scientific communities both locally and nationally. Several students graduated in the current reporting period with either Masters or doctoral degrees. Services: Several investigators served on local, national and international committees and expert panels in the areas of food safety, zoonotic diseases, bacterial pathogenesis, genomics, and emerging infectious diseases. Several investigators contributed to the editorship and/or Ad Hoc reviews of peer reviewed journal articles, white papers, and fact sheets Products: Number of novel findings was reported that addresses both the basic as well as applied needs of zoonotic and emerging infectious disease problems. Basic research has facilitated the development of control and preventive measures such as vaccines, therapeutics, and alternatives to antibiotics. Likewise, applied research has resulted in the development of improved and effective diagnostic and epidemiological surveillance strategies for the control of these diseases. During this period, we have isolated and stored various field and outbreak isolates of zoonotic pathogens such as influenza, and foodborne pathogens such as norovirus, Campylobacter, Shiga toxin producing E. coli, and Salmonella. Our efforts also established/expanded of newer collaborations among and across the other members of the regional committees nationally as well as with international peers. Several extension materials were generated containing information relevant to good agricultural practices, effective control and diagnostic methods. Dissemination: Several meetings were conducted with the stakeholders including livestock and poultry producers as well as veterinarians. Written and electronic media was distributed to stakeholders and research findings were published in peer reviewed journals. PARTICIPANTS: Veterinary Researchers Veterinarian Livestock producers Policy makers TARGET AUDIENCES: Livestock Producers and processors Scientists PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Change in Knowledge: Graduate and undergraduate students and postdoctoral fellows acquired research skills in the area of epidemiology, diagnostics, vaccine development, pre- and post harvest control strategies, as well as in basic research relating to mechanisms of disease pathogenesis. These trainings resulted in the completion of several Masters and Graduate students dissertation and endured them with skills to conduct independent research. Similarly researchers were trained in extension and education such that they can effectively communicate with different stakeholders. Several invention disclosures were filed during this period and some were awarded with patent for the novelty and its significance either to livestock industry or public health. Change in condition: The knowledge gained and communicated to different stakeholders is expected to enhance food safety, reduce risk of zoonoses, aid in the development of vaccines and therapeutic against human infections and preparedness against emerging and re-emerging infectious disease of livestock as well as public health significance. Patents issued are: Renukaradhya, GJ, Binjawadagi, B, Kang, O, Zimmerman, JJ,. "Development and Validation of Pig Oral Fluid Based Diagnostic Tool to Determine the Herd Immunity." filed 2012, (patent pending), and Application No. 61/730543. 2. Renukaradhya G J, Dwivedi V, Binjawadagi B, and Torrelles J B. Inventor. "Compositions and methods for treating and preventing Porcine Reproductive and Respiratory Syndrome." The Ohio State University. filed 2012, (patent pending), and Application No. 61637547. Marcus, PI, J.M. Ngunjiri, M.J. Sekellick, C.W.Lee. Influenza virus populations, methods of use and methods of making thereof. Filed 2012 (patent pending) and application no. UCT0184US2. Miller, S., G. Rajashekara, X. Xu. Clavibacter Michiganensis Subsp. Michiganensis Bioluminescent mutants and application. Patent no. US8227186B1, 2012.

Publications

  • 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 Pathog Dis. 2012. Vol. 10, (Dec 2012): PMID: 23259503.
  • Kassem II, Khatri M, Esseili MA, Sanad YM, Saif YM, Olson JW, Rajashekara G. 2012 "Respiratory proteins contribute differentially to Campylobacter jejuni's survival and in vitro interaction with hosts' intestinal cells." BMC Microbiol. Vol. 12, no. 1. (Nov 2012): PMID: 23148765.
  • 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." Poultry Science. Vol. 4, (Apr 2012): 1032-7.
  • Ali, A., Ibrahim, M., Eladl, A. H., Saif, Y.M. and Lee, C.W. 2012 Enhanced Replication of Swine Influenza Viruses in Dexamethasone-Treated Juvenile and Layer Turkeys. Vet Microbiol. 2012 S0378-1135(12)00546-9.
  • Ali, A., Khatri, M., Wang, L., Saif, Y.M. and Lee, C.W. 2012 Identification of Swine H1N2/Pandemic H1N1 Reassortant Influenza Viruses in Pigs, United States. Vet Microbiol. 2012. 158(1-2):60-8.
  • Ngunjiri JM, Lee CW, Ali A, Marcus PI 2012 Influenza virus interferon-inducing particle efficiency is reversed in avian and mammalian cells, and enhanced in cells co-infected with defective-interfering particles. 2012. 32(6):280-5.
  • Dwivedi V, C. Manickam, B. Binjawadagi, R. Patterson, D. Linhares, M.P. Murtaugh, and G.J. Renukaradhya (2012). Evaluation of immune responses to porcine reproductive and respiratory syndrome virus in pigs during early stage of infection. Virology Journal. 9:45. PMID: 22340040
  • Renukaradhya, G.J., V. Dwivedi, C. Manickam, B. Binjawadagi, D. Benfield (2012). Mucosal vaccines to control porcine reproductive and respiratory syndrome: a new perspective. Animal Health Research Reviews, 13(1):21-37. Cambridge University Press. PMID: 22717576
  • Broxmeyer H.E, Christopherson K, Hangoc G, Cooper S, Mantel C, Renukaradhya G. J, and Brutkiewicz R.R (2012). CD1d Expression on and regulation of murine hematopoietic stem and progenitor cells. Blood, 119(24): 5731-41. PMID: 22535665
  • Manickam C, V. Dwivedi, R. Patterson, K. Dodson and G.J. Renukaradhya (2013). Differential regulation of mucosal immune responses at various mucosal tissues in pigs infected with VR2332 strain of PRRSV. Vet. Microbiol., 162:68-77.
  • Manickam C, V. Dwivedi, J. Miller, T. Papenfuss and G.J. Renukaradhya (2012) Mycobacterium tuberculosis whole cell lysate enhances proliferation of CD8 positive lymphocytes and nitric oxide secretion in the lungs of live porcine respiratory and reproductive syndrome virus vaccinated pigs. Viral Immunol., 26(1),
  • Dwivedi V, C. Manickam, B. Binjawadagi, Joyappa, D, and G.J. Renukaradhya (2012). Biodegradable nanoparticle-entrapped vaccine induces cross-protective immune response against a virulent heterologous respiratory viral infection in pigs. PLOS One. 7(12):e51794.
  • Jackwood DJ 2012 Molecular epidemiologic evidence of homologous recombination in infectious bursal disease viruses. Avian Dis. Sep;56(3):574-7. PMID: 23050476
  • Jackwood DJ, Crossley BM, Stoute ST, Sommer-Wagner S, Woolcock PR, Charlton BR 2012 Diversity of genome segment B from infectious bursal disease viruses in the United States. Avian Dis. 2012 Mar;56(1):165-72. PMID: 22545543
  • Gelb J Jr, Jackwood DJ, Mundt E, Pope CR, Hein R, Slacum G, Harris JM, Ladman BS, Lynch P, Bautista DA, Ruano JM, Troeber MM 2012 Characterization of infectious bursal disease viruses isolated in 2007 from Delmarva commercial broiler chickens. Avian Dis. 2012 Mar;56(1):82-9. PMID: 22545532
  • Jackwood DJ 2011 Viral competition and maternal immunity influence the clinical disease caused by very virulent infectious bursal disease virus. Avian Dis. 2011 Sep;55(3):398-406. PMID: 22017037
  • Chang, K.O., L. J. Saif and Y. Kim. 2012. Reoviruses (Rotaviruses and Reoviruses). In: Diseases of Swine. 10th Ed (J.J. Zimmerman, et al, Eds.), Wiley-Blackwell, Ames. Iowa. pp 621-634.
  • Esseili MA, Wang Q, Zhang Z, Saif LJ. 2012 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
  • 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. 2012. 78:786-94.
  • 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-α on infectivity of human norovirus using a gnotobiotic pig model for the study of antivirals. PLoS One 7:e41619.
  • 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.
  • 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.
  • 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 157:1999-2003.
  • Rauf, A., M. Khatri, M.V. Murgia and Y.M. Saif 2012 Fas/FasL and perforin systems as important mechanisms of T cell-mediated cytotoxicity in infectious bursal disease virus infected chickens. Results in Immunology 2 :112-119.
  • Murgia, M.V., A. Rauf, Y. Tang, E. Gingerich, C.W. Lee and Y.M. Saif 2012: Prevalence of parvoviruses in commercial turkey flocks. Avian Dis. 56 :744-749.
  • Harris LJ, Bender J, Bihn EA, Blessington T, Danyluk MD, Delaquis P, Goodridge L, Ibekwe AM, Ilic S, Kniel K, Lejeune JT, Schaffner DW, Stoeckel D, Suslow TV 2012 A Framework for Developing Research Protocols for Evaluation of Microbial Hazards and Controls during Production That Pertain to the Quality of Agricultural Water Contacting Fresh Produce That May Be Consumed Raw. J Food Prot. 2012 Dec;75(12):2251-73. PMID: 23212026
  • Cernicchiaro N, Pearl DL, McEwen SA, Harpster L, Homan HJ, Linz GM, Lejeune JT 2012 Association of wild bird density and farm management factors with the prevalence of E. coli O157 in dairy herds in Ohio (2007-2009). Zoonoses Public Health. 2012 Aug;59(5):320-9. PMID: 22333436
  • Saif, L.J., M.P. Pensaert, K. Sestak, S.G. Yeo, K. Jung 2012 Coronaviruses. In Diseases of Swine (J.J. Zimmerman, et al. Eds.), 10th Edition, Wiley-Blackwell, Ames, Iowa. pp 501-524.


Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: Activities: In the current year we conducted a large number of hypothesis-driven experiments that are both basic as well applied. Findings from our research were presented in several National as well as international scientific meetings and published in peer reviewed journals. In addition, we also mentored large number of Graduate, Undergraduate, High School students and postdoctoral fellows in various aspects of Emerging and Re-emerging zoonotic diseases. Events: Results obtained from our research were presented in several local, national as well as international scientific meetings. Presentations were also targeted for different stakeholders including livestock producers, food processors and policy makers. Services: Several PIs served on local, national and international committees and expert panels in the areas of food safety, zoonotic diseases and emerging infectious diseases. Products: Large body of research findings were identified through basic research that would aid in the development of effective control strategies. Several diagnostic assays were developed, particularly to detect different subtypes of influenza viruses. Further, newer vaccines were developed and evaluated under laboratory experimental conditions, which will be evaluated in the field to make available for field applications. Besides we increased our repository of zoonotic pathogens. Large number research conducted during this period resulted in newer collaborations locally, nationally and internationally. Extension materials were prepared and communicated to different stakeholders. Dissemination: Several meetings were conducted with the stakeholders including livestock and poultry producers as well as veterinarians. Written and electronic media was distributed to stakeholders and research findings were published in peer reviewed journals. PARTICIPANTS: Veterinary Researchers Veterinarian Live stock producers Policy makers TARGET AUDIENCES: Livestock Producers and processors Scientists PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Change in Knowledge: Graduate and undergraduate students and postdoctoral fellows acquired research skills in the area of epidemiology, diagnostics, vaccine development, pre- and post harvest control strategies, as well as in basic research. These trainings resulted in the abilities of student to conduct research independently. Similarly extension educators have sufficient knowledge to effectively communicate with different stakeholders. Some of the research conducted resulted in patent applications and drawn attention from public for unique work done by our researchers. Change in condition: The knowledge gained and communicated to different stakeholders is expected to enhance food safety, reduce risk of zoonoses, aid in the development of vaccines and therapeutic against human infections and preparedness against emerging and re-emerging infectious disease of livestock as well as public health significance.

Publications

  • Rauf, A., M. Khatri, M.V. Murgia and Y.M. Saif. 2011 Expression of perforin-granzyme pathway genes in the bursa of infectious bursal disease virus-infected chickens. Dev Comp Immunol 35:620-627
  • Eladl, A.E., K.I. El-Azm, A.E. Ismail, A. Ali, Y.M. Saif and C.W. Lee. 2011 Genetic characterization of highly pathogenic H5N1 avian influenza viruses isolated from poultry farms in Egypt. Virus Genes. 43:272-80
  • Rauf, A., M. Khatri, M.V. Murgia, K. Jung, and Y.M. Saif. 2011 Differential modulation of cytokine, chemokine and Toll like receptor expression in chickens infected with classical and variant infectious bursal disease virus. Vet Res 42:85
  • Takanashi S, Wang Q, Chen N, Shen Q, Jung K, Zhang Z, Yokoyama M, Lindesmith LC, Baric RS, Saif LJ. 2011 Characterization of emerging GII.g/GII.12 noroviruses from a gastroenteritis outbreak in the United States in 2010. J Clin Microbiol. Sep;49(9):3234-44
  • Wang Q, Scheuer K, Ahang Z, Gebreyes WA, Molla BZ, Hoet AE, Saif LJ. 2011 Characterization and prevalence of a new porcine Calicivirus in Swine, United States. Emerg Infect Dis. Jun;17(6):1103-6
  • Vega C, Bok M, Chacana P, Saif L, Fernandez F, Parreno V. 2011 Egg yolk IgY: protection against rotavirus induced diarrhea and modulatory effect on the systemic and mucosal antibody responses in newborn calves. Vet Immunol Immunopathol. Aug 15;142(3-4):156-69
  • Matthijnssens J, Ciarlet M, McDonald SM, Attoui H, Banyai K, Brister JR, Buesa J, Esona MD, Estes MK, Gentsch JR, Iturriza-Gomara M, Johne R, Kirkwood CD, Martella V, Mertens PP, Nakagomi O, Parreno V, Rahman M, Ruggeri FM, Saif LJ, Santos N, Steyer A, Taniguchi K, Patton JT, Desselberger U, Van Ranst M. 2011 Uniformity of rotavirus strain nomenclature proposed by the Rotavirus Classification Working Group (RCWG).Arch Virol. Aug;156(8):1397-413
  • Park SI, Matthijnssens J, Saif LJ, Kim HJ, Park JG, Alfajaro MM, Kim DS, Son KY, Yang DK, Hyun BH, Kang MI, Cho KO. 2011 Reassortment among bovine, porcine and human rotavirus strains results in G8P[7] and G6P[7] strains isolated from cattle in South Korea. Vet Microbiol. Aug 26;152(1-2):55-66
  • Wen K, Li G, Zhang W, Azevedo MS, Saif LJ, Liu F, Bui T, Yousef A, Yuan L. 2011 Development of gamma/delta T cell subset responses in gnotobiotic pigs infected with human rotaviruses and colonized with probiotic lactobacilli. Vet Immunol Immunopathol. Jun 15;141(3-4):267-75
  • Vlasova AN, Halpin R, Wang S, Ghedin E, Spiro DJ, Saif LJ. 2011 Molecular characterization of a new species in the genus Alphacoronavirus associated with mink epizootic catarrhal gastroenteritis. J Gen Virol. Jun;92(Pt 6):1369-79
  • Jackwood DJ, Sommer-Wagner SE. 2011 Amino acids contributing to antigenic drift in the infectious bursal disease Birnavirus (IBDV). Virology. Jan 5;409(1):33-7
  • Jackwood DJ, Sommer-Wagner SE, Crossley BM, Stoute ST, Woolcock PR, Charlton BR. 2011 Identification and pathogenicity of a natural reassortant between a very virulent serotype 1 infectious bursal disease virus (IBDV) and a serotype 2 IBDV. Virology. Nov 25;420(2):98-105
  • Jackwood DJ. 2011 Viral competition and maternal immunity influence the clinical disease caused by very virulent infectious bursal disease virus. Avian Dis. Sep;55(3):398-406.
  • 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 Research Notes. Dec 30 ;4(1):570.
  • 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.
  • 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.
  • Gourapura J. Renukaradhya, Cordelia Manickam, Mahesh Khatri, Xiangming Li, Moriya Tsuji, Gireesh Rajashekara, and Varun Dwivedi. 2011 Functional invariant NKT cells in pig lungs regulate the airway hyperreactivity: a potential animal model. Journal of Clinical Immunology . 31(2):228-39
  • Williams M, Pearl D, and LeJeune J . 2011 Multiple-locus variable-nucleotide tandem repeat subtype analysis implicates European starlings as biological vectors for Escherichia coli O157:H7 in Ohio, USA J Appl Microbiol 77.1365-2672
  • Rodriguez-Palacios A, Loerch. 2011 Transient fecal shedding and limited animal-to-animal transmission of Clostridium difficile in naturally infected feedlot cattle. Appl Environ Microbiol 77: 3391-3397
  • Rodriguez-Palacios A, LeJeune, JT. 2011 Moist heat resistance, spore aging, and superdormancy in Clostridium difficile. Appl Environ Microbiol. DOI:10.1128 AEM.01589-10
  • Meng J, LeJeune J, Zhao T. 2011 Enterohemorrhagic Escherichia coli In Fundamentals of Food Microbiology 4th Edition
  • Rodriguez-Palacios A, Koohmaraie M, and LeJeune J. 2011 Prevalence, Enumeration, and Antimicrobial Agent Resistance in Clostridium difficile in Cattle at Harvest in the United States. JFP-11-141
  • Westphal A, Williams M, Baysal-Gurel F, LeJeune JT, and McSpadden-Gardener B. 2011 General Suppression of Escherichia coli O157:H7 in sand-based dairy livestock bedding. Appl Environ Microbiol. 77:2114-2121
  • Casagrande G, LeJeune J, Belury MA, Medeiros LC. 2011 Registered dietitian's personal beliefs and characteristics predict their teaching or intention to teach fresh vegetable food safety. Appetite 56:469-475
  • Yassine HM, Lee CW, Saif YM. 2011 Interspecies Transmission of Influenza A Viruses Between Swine and Poultry. Curr Top Microbiol Immunol. 2011. Dec 14
  • Ali A, Daniels JB, Zhang Y, Rodriguez-Palacios A, Hayes-Ozello K, Mathes L, Lee CW. 2011 Pandemic and Seasonal Human Influenza Virus Infections in Domestic Cats: Prevalence, Association with Respiratory Disease, and Seasonality Pattern. J Clin Microbiol 49(12):4101-5
  • Wang L, Qin Z, Pantin-Jackwood M, Faulkner O, Suarez DL, Garcia M, Lupiani B, Reddy SM, Saif YM, Lee CW. 2011 Development of DIVA (differentiation of infected from vaccinated animals) vaccines utilizing heterologous NA and NS1 protein strategies for the control of triple reassortant H3N2 influenza in turkeys. Vaccine. 29(45):7966-74
  • Eladl AE, El-Azm KI, Ismail AE, Ali A, Saif YM, Lee CW. 2011 Genetic characterization of highly pathogenic H5N1 avian influenza viruses isolated from poultry farms in Egypt. Virus Genes. 43(2):272-80
  • Issmat I. Kassem, Qijing Zhang, Gireesh Rajashekara. 2011 The twin arginine translocation (Tat) system : Contributions to the pathobiology of Campylobacter jejuni. Future Microbiol. Nov;6(11):1315-27
  • Issmat I. Kassem, and Gireesh Rajashekara. 2011 An ancient molecule in a recalcitrant pathogen: the contributions of Inorganic polyphosphate (Poly-P) to the pathogenesis and stress responses of Campylobacter jejuni. Future Microbiol. Oct;6:1117-20
  • Qin Z, Clements T, Wang L, Khatri M, Pillai SP, Zhang Y, Lejeune JT, Lee CW. 2011 Detection of influenza viral gene in European starlings and experimental infection. Influenza Other Respi Viruses. 5(4):268-75
  • Yassine HM, Khatri M, Lee CW, Saif YM. 2011 Potential role of viral surface glycoproteins in the replication of H3N2 triple reassortant influenza A viruses in swine and turkeys. Vet Microbiol. 148(2-4):175-82
  • Khandelwal, A., G. J. Renukaradhya, M. Rajasekhar, G. Lakshmi Sita, and M. S. Shaila. 2011 Immune responses to Hemagglutinin-Neuraminidase protein of peste des petits ruminants virus expressed in transgenic peanut plants in sheep. Vet. Immunol. Immunopathol. 140(3-4):291-296
  • Dwivedi V, C. Manickam, R. Patterson, K. Dodson, and G.J. Renukaradhya. 2011 Intranasal delivery of whole cell lysate of Mycobacterium tuberculosis induces protective immune responses to a modified live porcine reproductive and respiratory syndrome virus vaccine in pigs. Vaccine 29(23):4067-76
  • Dwivedi V, C. Manickam, R. Patterson, K. Dodson, M. Murtaugh, J.B. Torrelles, L.S. Schlesinger, and G.J. Renukaradhya. 2011 Cross-protective immunity to porcine reproductive and respiratory syndrome virus by intranasal delivery of a live virus vaccine with a potent adjuvant. Vaccine 29(23):4058-66
  • Binjawadagi B, V. Dwivedi, C. Manickam, J.B. Torrelles, and G.J. Renukaradhya. 2011 Intranasal delivery of an adjuvanted modified live porcine reproductive and respiratory syndrome virus vaccine reduces the ROS production. Viral Immunol. 24(6):1-8


Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: Activities: In the current year a large number of hypothesis-driven experiments were conducted. Research findings presented in scientific meetings and published in peer reviewed journals. Besides, we also mentored large number of Graduate, Undergraduate and High School students as well as postdoctoral fellows in various aspects of Emerging and Re-emerging zoonotic diseases. Events: Our research findings were presented in several local, national as well as international scientific meetings. Presentations were also targeted for different stakeholders including livestock producers, food processors and policy makers. Services: Several PIs served on local, national and international committees and expert panels in the areas of food safety, zoonotic diseases and emerging infectious diseases. Products: Large body of research findings were identified through basic research that would in the development of effective control strategies. Several diagnostic assays were developed, particularly to detect different subtypes of influenza viruses. Further, newer vaccines were developed and evaluated under laboratory experimental conditions, which will be evaluated in the field to make available for field applications. Besides we increased our repository of zoonotic pathogens. Large number studies conducted during this period resulted in newer collaborations locally, nationally and internationally. Extension materials were prepared and communicated to different stakeholders. Dissemination: Several meetings were conducted with the stakeholders including livestock and poultry producers as well as veterinarians. Written and electronic media was distributed to stakeholders and research findings were published in peer reviewed journals. PARTICIPANTS: Veterinarian Veterinary researchers Live stock producers TARGET AUDIENCES: Livestock producers and processors PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Change in Knowledge: Graduate and undergraduate students and postdoctoral fellows acquired research skills in the area, epidemiology, diagnostics, vaccine development, pre- and post harvest control strategies, as well as in the areas basic research. These trainings resulted in the abilities of student conduct research independently. Similarly extension educators have sufficient knowledge to effectively communicate with different stakeholders. Some of the research conducted resulted in patent applications and drawn attention from public for unique work done by our researchers. Change in condition: The knowledge gained and communicated to different stakeholders is expected to enhance food safety, reduce risk of zoonoses and preparedness against emerging and re-emerging infectious disease of livestock as well as public health significance.

Publications

  • Marcus PI, Ngunjiri JM, Sekellick MJ, Wang L, Lee CW. In Vitro Analysis of Virus Particle Subpopulations in Candidate Live-Attenuated Influenza Vaccines Distinguishes Effective from Ineffective Vaccines. Journal of Virology. 84(21):10974-81. 2010.
  • Yassine HM, Khatri M, Lee CW, Saif YM. Potential role of viral surface glycoproteins in the replication of H3N2 triple reassortant influenza A viruses in swine and turkeys. Vet Microbiol. In Press, 2010.
  • Yassine HM, Khatri M, Lee CW, Saif YM. Characterization of an H3N2 triple reassortant influenza virus with a mutation at the receptor binding domain (D190A) that occurred upon virus transmission from turkeys to pigs. Virology Journal. 7(1):258. 2010.
  • Pillai SPS, Pantin-Jackwood M, Suarez DL, Saif YM , Lee CW. Pathobiological characterization of low pathogenicity H5 avian influenza viruses of diverse origins in chickens, ducks and turkeys. Arch Virol. 155(9): 1439-51. 2010.
  • W Cha, Y Ma, YM Saif, Lee CW. Development of microsphere-based multiplex branched DNA assay for the detection and differentiation of avian influenza virus. J Clin Microbiol. Vol. 7, no. 48: 2575-2577. 2010.
  • Pillai SPS, Saif YM, Lee CW. Detection of influenza A viruses in eggs laid by infected turkeys. Avian Dis. 54(2):830-3. 2010.
  • Pillai SPS & Lee CW. Species and age related differences in the type and distribution of influenza virus receptors in different tissues of chickens, ducks and turkeys. Virol J. 7:5. 2010.
  • Wang L, Yassine HM, Saif YM, Lee CW. Developing Live Attenuated Avian Influenza Virus In Ovo Vaccines for Poultry. Avian Dis. 54:297-301, 2010.
  • Pillai SPS, Pantin-Jackwood M, Yassine HM, Saif YM, Lee CW. The high susceptibility of turkeys to Influenza viruses of different origins implies their importance as potential intermediate hosts. Avian Dis. 54:522-526, 2010.
  • Avellaneda G, Mundt E, Lee CW, Jadhao S, Suarez DL. Differentiation of Infected and Vaccinated Animals (DIVA) Using the NS1 Protein of Avian Influenza Virus. Avian Dis. 54:278-286. 2010.
  • Avellaneda G, Lee CW, Suarez DL. A Heterologous Neuraminidase Subtype Strategy for the Differentiation of Infected and Vaccinated Animals (DIVA) for Avian Influenza Virus Using an Alternative Neuraminidase Inhibition Test. Avian Dis. 54:272-277. 2010.
  • Dharanesh Gangaiah, Zhe Liu, Jesus Arcos, Issmat I. Kassem, Yasser Sanad, Jordi B. Torrelles and Gireesh Rajashekara. 2010. Polyphosphate Kinase 2: A Novel Determinant of Stress Responses and Pathogenesis in Campylobacter jejuni. PLoS One, Aug 17;5(8). pii: e12142.
  • 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. 2010 Jul 18. [Epub ahead of print}.
  • Gourapura J. Renukaradhya, Cordelia Manickam, Mahesh Khatri, Xiangming Li, Moriya Tsuji, Gireesh Rajashekara, and Varun Dwivedi. 2010. Functional invariant NKT cells in pig lungs regulate the airway hyperreactivity: a potential animal model. Journal of Clinical Immunology (In Press).
  • Drozd M, Kassem II, Gebreyes W, Rajashekara G. 2010. A quantitative polymerase chain reaction assay for detection and quantification of Lawsonia intracellularis. J Vet Diagn Invest. Mar;22(2):265.


Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: Output: Activities: During the current year we conducted a large number of hypothesis-driven experiments, data were analyzed and presented in scientific meetings and published in peer reviewed journals. Graduate, Undergraduate and High School students as well as postdoctoral fellows were mentored. Events: Presentation on original research and on issues related to food safety and zoonotic diseases were presented locally, nationally, and internationally. These presentations were directed towards different stakeholders including livestock producers and food processors. Besides scholarly research findings were presented in research oriented scientific meetings. Services: Several PIs served on local, national and international committees and expert panels in the areas of food safety and zoonotic diseases. Products: Several diagnostic and are therapeutics including vaccines were developed and evaluated in the laboratory which will need further study for field applications. Besides we increased our repository of zoonotic pathogens. Collaborations locally, nationally and internationally were established. Extension materials were prepared and communicated to different stakeholders. Dissemination: Several meetings were conducted with the stakeholders and veterinarians. Written and electronic media was distributed to stakeholders and research findings were published in peer reviewed journals. PARTICIPANTS: Veterinarians, veterinary researchers, animal producers TARGET AUDIENCES: Animal producers and processers PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Outcomes/Impact Change in Knowledge: Graduate and undergraduate students and postdoctoral fellows acquired research skills and are capable of conducting research independently. Similarly extension educators have sufficient knowledge to effectively communicate with different stakeholders. Some of the research conducted resulted in patent applications and drawn attention from public for unique work done by our researchers. Change in condition: The knowledge gained and communicated to different stakeholders is expected to enhance food safety and zoonoses reduce the number of illnesses associated with food producing animals and their products.

Publications

  • Publication: Dharanesh Gangaiah, Issmat I. Kassem, Zhe Liu, and Gireesh Rajashekara. 2009. Importance of polyphosphate kinase 1 for Campylobacter jejuni viable-but-nonculturable cell formation, natural transformation, and antimicrobial resistance. Appl Environ Microbiol. 75:7838-49. Rajashekara, G, Drozd, M., Gangaiah, D., Jeon, B., Liu, Z., and Zhang, Q., 2009, Functional Characterization of The Twin-arginine Translocation System in Campylobacter jejuni. Foodborne pathogens and Dis. 6: 935-945. Dharanesh Gangaiah , Issmat I. Kassem, Byeonghwa Jeon, Zhe Liu, Qijing Zhang, and Gireesh Rajashekara. 2009. "Polyphosphate Kinase 1 is Important for VBNC Formation, Natural Transformation and Antimicrobial Resistance in Campylobacter jejuni." In: 15th International workshop on Campylobacter, Helicobacter and Related Organisms. September 2Toki Messe, Niigata, Japan. Dharanesh Gangaiah, B. Adhikari, B. Jeon, Q. Zhang , Z. Liu, Y. Sanad, M. Drozd, and G. Rajashekara. 2009. "Deciphering the role of Polyphosphate kinases in Campylobacter jejuni colonization/ pathogenesis." In: 146th Annual AVMA Convention. July 11Seattle, Washington, USA. Rajashekara, Gireesh. 2009. "Campylobacter jejuni stress responses and in vivo colonization: Relevance to food safety." In: International conference on Food, Environment, and Health. June 20, Xian, China. Lenz J, Joffe D, Kauffman M, Zhang Y, LeJeune J. Perceptions, practices, and consequences associated with foodborne pathogens and the feeding of raw meat to dogs. Can Vet J. 2009 Jun;50(6):637-43.PMID: 19721784 Cernicchiaro N, Pearl DL, Ghimire S, Gyles CL, Johnson RP, LeJeune JT, Ziebell K, McEwen SA. Risk factors associated with Escherichia coli O157:H7 in Ontario beef cow-calf operations. Prev Vet Med. 2009 Nov 1;92(1-2):106-15. Kersting AL, Medeiros LC, LeJeune JT. Zoonoses and the physicians' role in educating farming patients. J Agromedicine. 2009;14(3):306-11. Ha, T.P., H.J. Kim, L.J. Saif, Y.J. Jeong, H.H. Kim, H.J. Kwon, S.J. Park, and K.O. Cho. 2009. Sequence analysis of unusual P[7]G5 bovine rotavirus strains reveals evidence of interspecies transmission. J Clin Microbiol. 47:3329-32. Park. SI, D.H. Park, L.J. Saif, Y.J. Jeong, D.J. Shin, Y.H. Chun, S.J. Park, H.J. Kim, M. Hosmillo, H.J. Kwon, M.I. Kang and K.O. Cho. 2009. Development of SYBR green real-time RT-PCR for rapid detection, antitation and diagnosis of unclassified bovine enteric calicivirus. J. Virol. Methods 159:64-68.


Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: Activities: A large number of hypothesis-driven experiments were conducted and data analyzed. Graduate students, undergraduate students were mentored. Events: Presentations on original research, and on over issues related to food safety and zoonotic disease were presented locally, nationally, and internationally. Many presentations targeted food producers while others were scientifically-orientated and directed at other researchers. Services: Several of the PIs served on national and international committees and Expert panels. Products: Isolate banks of zoonotic pathogens were expanded. Networks and collaborations were further developed. Information Extension booklets for vegetable producers was expanded to include food safety information. A manual for prudent antibiotic use in dairy productions was produced. A website www.oardc.osu.edu/getsmart was developed. Dissemination: Meetings were held for vegetable producers and other for veterinarians. Written and electronic media was disseminated to program participants and send via direct mailing to targeted audiences. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Change in knowledge: Extension personnel have a better understanding of knowledge gaps and preferred information dissemination routes of farmers and producers. They now have improved skills to communicate agricultural risk to stakeholders. Producers have better tools for decision -making. Change in actions: Baseline levels of knowledge, skill and behavoir were measured. One group of growers self-reported a change in management behavoir (use of fresh and incompletely composted manure on vegetables). Change in conditions: The change in vegetable fertilization practices is expected to increase food safety and reduce the number of illnesses associated with microbiologically contaminated produce.

Publications

  • Aruscavage, D., Miller, S., Lewis-Ivey, M., Lee, K., and LeJeune, J. (2008) Survival and dissemination of Escherichia coli O157:H7 on physically and biologically damaged lettuce plants. J Food Protect. 71:2384-2388.
  • Ilic, S., Odomura, J., and LeJeune, J. (2008) Coliforms and prevalence of generic Escherichia coli and foodborne pathogens on minimally processed spinach in two packing plants. J Food Protect. 71:2398-2403.
  • Wilson, R, Tucker, M, Hooker, N, LeJeune, J. and Doohan, D. (2008) Perceptions and beliefs about weed management: Perspectives of Ohio grain and produce farmers. Weed Technology 22:339-350.
  • LeJeune, J. T., Homan, J., Lenz, G., and Pearl, D. (2008). Role of European Starlings in the Transmission of E. coli O157 on Dairy Farms. Proc 23rd Vetebr Pest Conf. (P31-35).


Progress 01/01/07 to 12/31/07

Outputs
OUTPUTS: Multi-disciplinary (Food Science, Veterinary Medicine, Bacterial Ecology, Plant Pathology, Horticulture and Crop Science, Human Behavoir, etc) teams were formed to address complex issues continually emerging in food safety. Activities included not only laboratory and field studies to achieve the stated goals, but also undergraduate, graduate, and professional teaching, graduate student mentoring and outreach events including workshops, symposia, and conference presentations. PARTICIPANTS: Individuals: LeJeune, J. T.; Saif, Y. M.; Saif, L.; Jackwood, D.; Theil, K. Kauffman, MD, Partner Organizations: Ohio Depts of Health and Ag Training or professional development:Graduate students: Dan Aruscavage, Sanja Ilic, Alex Rodriguez, Ann Kersting. TARGET AUDIENCES: Veterinarians. Public Health Officals and Agencies. Food Producers: livestoch and vegetable and fruit growers. Research Scientists: Health, Evironment. Public. PROJECT MODIFICATIONS: None

Impacts
Pathogens that contaminate the food supply have been better characterized. Specifically, these studies provide insight in to the emergence and evolution of pathogens in the food chain. For example, we have identified that antibiotic resistance genes may be transferred by bacteriophage among strains of Salmonella, that not Escherichia coli O157 are of equal pathogenic potential, and that management factors (antibiotic use, stocking density) on farms may impact the prevalence of Salmonella in pigs. In addition the distribution of pathogenic organisms in wildlife was reported

Publications

  • Nagy J.O, Zhang, Y., Yi, Liu, X., Motari, E., Jing Song, C., Lejeune, JT and Wang,G. (2007) Glycopolydiacetylene nanoparticles as a chromatic biosensor to detect Shiga-like toxin producing Escherichia coli O157:H7, Bioorg. Med. Chem. Lett. 18(2):700-3.
  • Zhang, Y and LeJeune, J (2007) Transduction of bla(CMY-2), tet(A), and tet(B) from Salmonella enterica subspecies enterica serovar Heidelberg to S. Typhimurium. Vet Microbiol. 2007 Dec 4 [Epub ahead of print]
  • Baker, D., Moxley, R, Steele, M., LeJeune J., Chistopher-Hennings, J., Chen., Hardwidge, P., and Francis, D.(2007) Virulence variation among Escherichia coli O157:H7 strains isolated from human disease outbreaks and healthy cattle Appl Environ Microbiol 73:7338-46
  • Malone, A, A. Yousef, and J. LeJeune (2007) Association of Prophage Antiterminator Q Alleles with in vitro processing treatments. J Food Protect .70:2617-9.
  • Wetzel, A and J. T. LeJeune (2007) Isolation of Escherichia coli O157:H7 strains that do not produce Shiga toxin from bovine, avian and environmental sources. Lett Appl. Micro 45:504-507
  • LeJeune, J (2007) The Scapegoat for Vegetable Safety. Food Prot Trends 9:728.
  • Funk,J, Wittum, T, LeJeune, J., Rajala-Schultz, P., and Mack, A. (2007). Evaluation of stocking density and subtherapeutic chlortetracycline on Salmonella enterica subsp. enterica shedding in growing swine. Vet Microbiol 124:202-208.
  • Jijon, S, Wetzel, A, LeJeune, J. (2007) Salmonella enterica isolated from Wildlife at two Ohio Rehabilitation Centers. J of Zoo Wildlife Medicine 38:409-413.
  • Bowman, A, Glendening, C, Wittum, T, LeJeune, J, Stich, R, and Funk. 2007. Prevalence of Yersinia enterocolitica in different phases of production on swine farms. J Food Prot. 70(1):11-6.
  • Wetzel, A and LeJeune J. (2007) Preharvest control of Escherichia coli O157 in cattle J Animal Sci. 85(13 Suppl):E73-80


Progress 01/01/06 to 12/31/06

Outputs
Objectives 1) Mechanisms of microbial pathogenesis of zoonotic agents, including the production of bacterial toxins 2) Immune Response using animal models of infection and immunity; 3) Epidemiology and natural history of infection; 4) Development of vaccines, therapeutics, adjuvants, and diagnostics for food safety and Biodefense. Both field studies and laboratory experiments have been conducted and are ongoing to achieve these aims. One study considered gene location within bacteria as a function of genetic element mobility. Our emphasis is on prophage encoding of bacterial virulence factors (VFs). At least four mechanisms potentially contribute to phage encoding of bacterial VFs: (i) Enhanced gene mobility; (ii) Epistatic interactions; (iii) Phage could amplify VFs during disease progression; and finally (iv) phage-encoded VFs could enhance phage Darwinian fitness, particularly by acting as ecosystem-modifying agents. Pathogens on edible plants present a significant potential source of human illness. From 1991 to 2002, 21% of Escherichia coli O157:H7 outbreaks were from produce-related sources. E. coliO157and other enteric bacteria can contaminate the surface of edible plants both pre- and postharvest. Some pathogens do not survive on the leaf surface or are removed by washing, but a significant portion of these enteric pathogens can persist on the surface and proliferate. Proliferation of these dangerous pathogens can increase the likelihood of foodborne disease associated with fresh or minimally processed produce. Several intrinsic and extrinsic factors determine the ability of enteric pathogens to attach and proliferate in the phyllosphere of plants. These include motility of the pathogen, leaching of nutrients by the plant, and interaction with epiphytic organisms. The interaction of enteric pathogens with the environment can lead to internalization into tissue, incorporation into biofilms, and genetic transfer. Current produce sanitation practices can reduce the microbial load from1 log10 to 3 log10, so there are many new treatments possible. Understanding the ecology of enteric pathogens on plants is important to the development of sanitation methods and biocontrol agents. This knowledge can also assist the farmer in preventing contamination. With increasing consumption and importation of produce, its safety is a high priority for processors and U.S. consumers. Food safety may be markedly improved with proper attention to pathogens on edible plants.

Impacts
Understanding the underlying microbiological and ecological factors driving the emergence and persistence of foodborne pathogens has provided targeted direction for applied research projects that address these principles.

Publications

  • Abedon, S.T., LeJeune, J.T. (2005). Why Bacteriophage encode exotoxins and other virulence factors. Evolutionary Bioinformatics Online 1:97-110. (http://www.la-press.com/evolbio.htm).
  • Funk, J, Wittum, T, Rajala-Schultz, P,. LeJeune, J (2006) The effect of subtherapeutic chlortetracycline on antimicrobial resistance in the fecal flora of swine. Microbial Drug Resistance 12:210-8.
  • Aruscavage, D Lee, K, Miller, S and LeJeune, JT. (2006) Food Safety Implications of Escherichia coli O157 and Salmonella spp. Present on Edible Plants. Journal of Food Safety 71:R89-R99.


Progress 01/01/05 to 12/31/05

Outputs
Controlled studies have elucidated the primary factors influencing the dissemination of Shiga-toxin encoding phages from E. coli O157. We have begun to test several commonly used therapeutic antibiotics (Micotil, Nuflor, Baytril, Naxcel, and Tetracycline) on their ability to induce phage in vitro. The positive control treatment (not shown in figure) with mitomycin C resulted in approximately 20x increase. Similar results were observed regardless of whether or not bile salts were included in the test medium. Again these results are still preliminary and statistical analysis has not been conducted to determine significance of differences between treatments, however, results suggest (see figure) that at minimum inhibitory concentration of each antibiotic, these agents induce phage release only marginally, if at all. In order to more precisely predict food safety risks, the fecal presence of foodborne pathogens among animals at slaughter must be correctly determined. Quantification of E. coli O157 is also desirable. In two separate experiments, detection and enumeration of a nalidixic acid resistant strain of E. coli O157 in bovine feces was assessed by culture on MacConkey agar supplemented with nalidixic acid (MACnal) and compared to overnight broth enrichment followed by immunomagnetic separation (IMS) and to direct plating (DP) of dilutions of bovine feces onto Sorbitol MacConkey agar containing cefixime and tellurite (SMACct). The sensitivity of detection of E. coli O157 by both DP and IMS was highly dependent upon the initial concentration of the target organism in the sample. Sensitivity of detection by IMS was poor below 100 CFU/g, but was better, and not affected by initial E. coli O157 numbers, above this concentration. Sensitivity of detection of E. coli O157 in bovine feces at low initial concentrations is very poor for both direct plating and IMS. Direct plating of dilutions of bovine feces on SMACct can be used to determine the magnitude of fecal E. coli excretion among cattle excreting greater than 100 CFU/g. Among positive samples identified by direct plating on SMACct, the direct counts of E. coli O157:H7 were highly correlated with the estimates obtained on the MACnal plates (r = 0.88, P<0.001). Because the majority of cattle excrete less than 102 CFU E. coli O157/g feces, most studies, including those using IMS methods, probably grossly underestimate the prevalence of E. coli O157 in cattle.

Impacts
The cause of the emergence of E. coli O157 as an important human pathogen remains enigmatic. The data presented in our study suggest that the use of therapeutic antibiotics in cattle contributes minimally to the release of stx-encoding phages and therefore has little impact on the emergence of novel Shiga toxin-producing E. coli strains. Our comparison of diagnostic assays is important because it demonstrates that direct plating for E. coli O157 can give reliable information about the prevalence of the organism in bovine feces. This direct plating assay, when used in pre-harvest food safety experiments will allow for the inexpensive, yet qualitative, assessment of E. coli in bovine feces, information that is critically needed to fill gaps in quantitative risk assessment models related to food and environmental safety.

Publications

  • LeJeune, J.T. and K. Dodson. 2005. Prevalence of Salmonella spp., Campylobacter jejuni and E. coli O157 in cull dairy cattle marketed in Northeast Ohio, USA. J Food Protect 68:927-931.


Progress 01/01/04 to 12/31/04

Outputs
Progress: Objectives 1) Mechanisms of microbial pathogenesis of zoonotic agents, including the production of bacterial toxins 2) Immune Response using animal models of infection and immunity; 3) Epidemiology and natural history of infection; 4) Development of vaccines, therapeutics, adjuvants, and diagnostics for food safety and Biodefense. Both field studies and laboratory experiments have been conducted and are ongoing to achieve these aims. Controlled studies have elucidated the primary factors influencing the magnitude of Shiga-toxin production from E. coli O157. The immune response to enteric viruses and virus-like particles continues to be evaluated in gnotobiotic piglet model of human disease. Initial information is now available on the global distribution of pathogenic strains of E. coli O157, as well as farm-to-farm variability of Mycobacterium paratuberculosis strains, including isolates obtained from human cases of Crohn's Disease.

Impacts
Better diagnostic tools are now available for the characterization of E. coli O157:H7 and Mycobacterium avaium subsp. paratuberculosis. These test will be available to all researchers to expedite further research and understanding of these important bacterial agents

Publications

  • LeJeune JT, Abedon ST, Takemura K, Christie NP, Sreevatsan S. Human Escherichia coli O157:H7 genetic marker in isolates of bovine origin. Emerg Infect Dis [serial on the Internet]. 2004 Aug [date cited]. Available from: http://www.cdc.gov/ncidod/EID/vol10no8/03-0784.htm
  • LeJeune JT and Christie P (2004) Microbiological Quality of Ground Beef from Conventionally-Reared Cattle and ''Raised without Antibiotics'' Label Claims. J Food Prot. 67:1433-1473.
  • A. H. Ghadiali, M. Strother, N. E. Theus, R. W. Stich, B. Byrum, W. P. Shulaw, and S. Sreevatsan. Rapid detection and strain typing of Mycobacterium avium subsp. paratuberculosis from broth enriched cultures. J. Clin. Microbiol. 2004.
  • K. Takemura, M. Khadre, D. Joseph, A. Yousef, and S. Sreevatsan. An overview of ransmissible spongiform encephalopathies. Animal Health Research Reviews. In Press. 2004.
  • A. H. Ghadiali, S. A. Naser, E. J. B. Manning, and S. Sreevatsan. Short sequence repeats analysis of Mycobacterium avium subspecies paratuberculosis isolates from Crohn's disease patients exhibit genetic similarity to extant clones of bovine or caprine origin. J. Clin. Microbiol. 42(11):5345-5348. 2004.
  • N. E. Ramirez, L. A. Ward, and S. Sreevatsan. A review of the biology and epidemiology of cryptosporidiosis in humans and animals. Microbes and Infection. 6: 773-785. 2004.
  • A. S. Motiwala, A. Amonsin, M. Strother, E. J. B. Manning, V. Kapur, and S. Sreevatsan. Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis Recovered from Wild Life Species. J. Clin. Microbiol. 42(4):1703-1712. 2004.
  • A. Amonsin, L. L. Li, Q. Zhang, A. S. Motiwala, S. Sreevatsan, and V. Kapur. A Multi-Locus Short Sequence Repeat Sequencing Approach for Mycobacterium avium subsp. paratuberculosis strain differentiation. J. Clin. Microbiol. 42(4): 1698-1702. 2004.


Progress 01/01/03 to 12/31/03

Outputs
Objectives 1. Mechanisms of microbial pathogenesis of zoonotic agents, including the production of bacterial toxins 2. Immune Response using animal models of infection and immunity 3. Epidemiology and natural history of infection 4. Development of vaccines, therapeutics, adjuvants, and diagnostics for food safety and biodefense Research has progressed in all objectives. Many projects encompass all objectives. For example the identification of a novel target for the identification of pathogenic Shiga toxin-producing E. coli has led to a greater understanding of the mechanisms of pathogenesis and the epidemiology of this pathogen in human and animal populations. Likewise, given the uncertainty of zoonotic potential of Mycobacterium avaium subsp. paratuberculosis, work has also been conducted on this organism. The potential for the cross-species transmission of M. avium paratuberculois and the relation ship between M. avium paratuberculois can now be determined. This information is critical in understanding the natural history of Johnes disease and the possible relationship of this agent in the etiologic of Chrones disease. Expected impact Better diagnostic tools are now available for the characterization of E. coli O157:H7 and Mycobacterium avaium subsp. paratuberculosis. These test will be available to all researchers to expedite further research and understanding of these important bacterial agents.

Impacts
Better diagnostic tools are now available for the characterization of E. coli O157:H7 and Mycobacterium avaium subsp. paratuberculosis. These test will be available to all researchers to expedite further research and understanding of these important bacterial agents

Publications

  • A. S. Motiwala, A. Amonsin, M. Strother, E. J. B. Manning, V. Kapur, and S. Sreevatsan. Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis Recovered from Wild Life Species. J. Clin. Microbiol. (In press). 2003.
  • A. Amonsin, L. L. Li, Q. Zhang, A. S. Motiwala, S. Sreevatsan, and V. Kapur. A Multi-Locus Short Sequence Repeat Sequencing Approach for Mycobacterium avium subsp. paratuberculosis strain differentiation. J. Clin.Microbiol. (In Press). 2003.
  • A. Ozbek, F. C. Michel, Jr., M. Strother, A. S. Motiwala, B. Byrum, W. Shulaw, C. G. Thornton, and S. Sreevatsan. Evaluation of Two Recovery Methods for detection of Mycobacterium avium subsp. paratuberculosis by PCR: Direct Dilution-Centrifugation and C18-Carboxypropylbetaine Processing. FEMS Microbiol. Letters 229:145-151. 2003.