Source: IOWA STATE UNIVERSITY submitted to
NOVEL AND PRACTICAL APPROACHES FOR MITIGATION OF CAMPYLOBACTER IN POULTRY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
0228913
Grant No.
2012-68003-19679
Cumulative Award Amt.
$2,499,558.00
Proposal No.
2011-06515
Multistate No.
(N/A)
Project Start Date
Jul 15, 2012
Project End Date
Jul 14, 2018
Grant Year
2015
Program Code
[A4161]- Food Safety: Prevention and Control of Salmonella and Campylobacter in Poultry Flocks and Poultry Products, including Eggs
Project Director
Zhang, Q.
Recipient Organization
IOWA STATE UNIVERSITY
S. AND 16TH ELWOOD
AMES,IA 50011
Performing Department
Veterinary Medicine
Non Technical Summary
Campylobacter is a leading cause of foodborne illness in the U.S. and contaminated poultry meat is the leading source of human campylobacteriosis. Therefore, control of Campylobacter in poultry will have a significant impact on ensuring food safety. Despite prior research efforts, there are still no effective and practical interventions that can be applied by poultry industries to control Campylobacter. To address this important food safety problem, we have built an integrated project to evaluate novel and practical intervention strategies and to disseminate science-based knowledge to producers, processors and consumers, facilitating control of Campylobacter at both the production and consumption stages. Specifically, we will develop and evaluate preharvest interventions that reduce Campylobacter colonization in poultry; identify critical control points and effective treatments in poultry processing plants for reducing Campylobacter contamination of poultry carcasses; facilitate poultry industry (producers and processors) to implement effective intervention strategies and educate consumers for proper handling and cooking of poultry meat. The proposed work is developed with significant input from stakeholders and directly addresses the food safety priority areas set by USDA-NIFA. The planned activities utilize integrated and systems-based approaches and are expected to generate practical and effective intervention strategies to improve poultry meat safety and reduce the public health burden. The activities will be conducted by a team of highly experienced investigators and is expected to yield important outcomes that will significantly improve food safety and benefit U.S. poultry industry.
Animal Health Component
80%
Research Effort Categories
Basic
20%
Applied
80%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7123220117050%
7123260110050%
Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
3220 - Meat-type chicken, live animal; 3260 - Poultry meat;

Field Of Science
1100 - Bacteriology; 1170 - Epidemiology;
Goals / Objectives
The main goal of this application is to develop and evaluate effective and practical intervention strategies to control Campylobacter contamination of poultry meat and its transmission to consumers. The specific objectives are 1) to develop and evaluate pre-harvest interventions that reduce Campylobacter colonization in poultry flocks, 2) to identify the critical control points and effective treatments in poultry processing plants for reducing Campylobacter contamination of poultry carcasses, and 3) to facilitate poultry industries to adopt effective intervention strategies and educate consumers to avoid infection by Campylobacter. The project is developed with significant input from stakeholders and takes advantage of collaboration with three poultry companies. This is a 5-year-long project. Specific objective 1 and 3 will be conducted during the entire project period, while specific objective 2 will be carried out during the last three years. Once completed, we expect that 1) on-farm management practices will be successfully identified and implemented to reduce Campylobacter prevalence on broiler farms, 2) protective vaccines against Campylobacter colonization in chickens will be developed, 3) effective litter treatment methods for reducing Campylobacter will be identified, 4) the points of contamination of Campylobacter in processing plants will be identified and interventions at the highest risk points will be evaluated, 5) the gained information will be disseminated to industries to design effective in-plant Campylobacter reduction schemes, 6) the targeted stakeholders and consumers will gain awareness and knowledge regarding prevention and control of Campylobacter, and 7) positive impacts on improving food safety and public health will be achieved by reducing the transmission of Campylobacter via poultry meat.
Project Methods
Various microbiological, epidemiological, and molecular approaches will be used in this study. 1) A large-scale 2-year longitudinal study will be conducted to determine Campylobacter prevalence and the associated on-farm risk factors on poultry flocks. Cecal samples will be obtained from different broiler farms in multiple states throughout the 2-year period. Campylobacter presence will be determined via standard culture methods. Flock level and farm level-specific standards and procedures, health status, environmental conditions and management practices will be recorded via observation and questionnaires. Logistic regression analyses will be performed for relating Campylobacter colonization status to farm specific information. 2) Three vaccine regimens that have potential for mass vaccination on poultry farms will be evaluated: in ovo DNA vaccination, oral live Salmonella-vectored vaccines, and intranasal immunization with chitosan encapsulated subunit vaccines. 3) The effectiveness of aluminum sulfate and sodium bisulfate treatments of litter on Campylobacter survival in the litter will be evaluated by conducting laboratory experiments, controlled pen studies and on-farm treatments. 4) A process-mapping at commercial broiler processing plants will be carried out to document the points of introduction, reduction and amplification of Campylobacter in poultry during processing. The most promising interventions associated with Campylobacter reduction in carcasses will be further investigated in a pilot processing plant. 5) Research findings will be disseminated to targeted stakeholders including poultry producers and processors using multiple delivery methods such as internet, written communications, short training courses, symposiums, regulatory agency webinars, and presentations at meetings and conferences. 6) Consumers will be educated for proper cooking and handling of poultry meat by delivering educational messages, which is aimed to improve awareness and knowledge of Campylobacter and to control the pathogen by improved cleaning, sanitizing, and cooking behaviors.

Progress 07/15/12 to 07/14/18

Outputs
Target Audience:Poultry producers and processors, consumers, veterinarians, scientists, students. Changes/Problems:The only major change during the entire project was the location of the meat safety training short course. The short course was originally planned to be held at Iowa State University. To increase the impact of the workshop, the course was offered at the University of Georgia. The course was organized by Dr. Manpreet Singh at Georgia in collaboration with Dr. Jim Dickson at ISU. Dr. Singh is a professor at the Department of Poultry Science and Extension, and is a well-known expert on food safety, poultry processing, and Campylobacter. He also has extensive connections with the poultry industry. What opportunities for training and professional development has the project provided?Nine undergraduate, six graduate students, and two postdocs were trained under this project. How have the results been disseminated to communities of interest?This project had a dedicated component on extension and educational programs. The information generated from this project and other relevant information from various sources were aggressively disseminated to the stakeholders as outlined previously under the heading "Extension and educational outreach programs" above. In addition, findings from this project were presented at national and international conferences and published in peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?The project has been completed and this is the final report.

Impacts
What was accomplished under these goals? 1. Longitudinal prevalence and on-farm risk factor study. A longitudinal study was conducted to identify the on-farm risk factors associated with variation in Campylobacter prevalence in the U.S. commercial broiler production. Cecal contents and boot swabs were collected from 5-6 week-old birds from 406 conventional broiler flocks reared 15 farms for up to 8 consecutive production cycles. Prevalence rates of Campylobacter at the farm, house, and flock levels are found to be 93%, 79%, and 47%, respectively. There was considerable variation in prevalence among farms, houses and flocks. Importantly, Campylobacter prevalence varied remarkably among different farms, houses and flocks, with some houses/farms testing consistently negative while others being positive all the time over the entire sampling period. A risk factor analysis indicated that Campylobacter positivity/high prevalence was associated with increased feed conversion, increased adjusted prime cost, and decreased paw quality/increased percent burnt. 2. Vaccine development. By targeting CfrA and CmeC, two promising vaccine candidates, live attenuated Salmonella-vectored vaccines were developed and evaluated. However, oral vaccination of chickens failed to trigger significant systemic and intestinal mucosal immune responses and consequently did not confer protection against C. jejuni colonization chickens. By targeting CfrA and CmeC, we also explored and evaluated a new vaccination strategy, which combined the in ovo vaccination route with a DNA vaccine. However, single in ovo injection of the DNA vaccine at 18th day of embryonation failed to trigger significant IgG and IgA immune responses and did not confer protection against C. jejuni colonization in the intestine of chickens. To develop subunit vaccines for intranasal or oral vaccination, the purified rCfrA and rCmeC were encapsulated into chitosan microsphere. The encapsulated pCAGGS-CmeC and pCAGGS-CfrA DNA vaccines were also produced. However, none of the intranasal subunit vaccines induced immune response and protection against Campylobacter in chickens. 3. Litter treatments. Chemical formulations amendment regimes were tested to reduce broiler litter's moisture, pH and Campylobacter jejuni loads. The litter management experiments examined sodium bisulfate (SOB), ammonium sulfate (AS) and magnesium sulfate (MS), individually and in combination. In trial I, orally challenged broilers (with C. jejuni) were placed in rooms with different litter treatments. In trial 2, uninfected birds were placed into rooms containing C. jejuni-contaminated litter that was treated with different chemicals. In trial 3, broiler chickens (including uninfected and C. jejuni-inoculated seeder birds) were placed into rooms containing litter treated with different chemicals. The best treatment was found to be the inclusion of all 3 chemicals together. Furthermore, the combination of the 3 chemicals was effective in significantly reducing the colonization of chickens and litter with Campylobacter in trial II. Treatment of the litter did not significantly reduce the load of C. jejuni in experimentally inoculated chickens or in litter (trial I); however, it significantly protected chickens from being colonized with C. jejuni, and the litter remained free of C. jejuni (trial II), which was especially evident when using the AS + SOB + MS treatment. Finally, even though treatment of the litter did not prevent transmission of C. jejuni from orally inoculated seeder chickens to un-inoculated chickens, the litter was Campylobacter-free under the condition (trial III). I4. Processing plant study for postharvest control of Campylobacter We examined the contamination and distribution of Campylobacter on broiler processing lines for a series of poultry processing plants. A total of nine different processing plants were included in the analysis. Samples were collected from the plants on three continuous production days each week for a period of 6 weeks. Carcass sampling occurred at sites that were determined by the processing plant and reflected points that were part of their routine analysis. Of the total 2,030 carcass rinse samples obtained from 9 different sites along the process line at 9 plants, 580 (28.6%) were positive for Campylobacter spp., with the majority (90.5%) yielding C. jejuni. The majority of samples that were positive for Campylobacter occurred early in the process line (i.e. pre and post scald, and the rehang stage), and detection beyond the final dip was minimal at best. These data suggest that interventions at the later stages of processing have a significant impact in reducing the overall final carcass contamination by Campylobacter. 5. Extension and Educational Outreach Programs a. Internet website. A dedicated website (http://www.campypoultry.org/) provided a simple, yet efficient and comprehensive platform for stakeholders and the public to access information on Campylobacter and food safety. b. Educational and outreach programs for poultry industry. A variety of educational tools were developed to promote the message of the risks and intervention measures needed to reduce Campylobacter on the farm and at the processing plant. All of the materials are available for free access and download on the http://www.campypoultry.org/ and Center for Food Security and Public Health poultry diseases and resources website http://www.cfsph.iastate.edu/Species/poultry.php. We worked closely with the poultry producers and processors for dissemination of knowledge gained from this project and relevant literature into ready-to-use information that will be relevant for live production (preharvest) and processing (postharvest). On-site workshops were conducted by poultry extension specialists to generate awareness on the topic and provide a platform for thought-provoking discussions to improve intervention technologies and strategies that are already implemented in these operations. c. Meat safety training workshop. A 2.5-day workshop was conducted as part of the outreach and extension effort of the project. This workshop "Campylobacter workshop: Managing the performance standards for poultry" consisted of discussions about the USDA-FSIS performance standards for Campylobacter in poultry and strategies to manage the control of this pathogen during the pre- and post-harvest stage of production. d. Consumer education. The objectives were to improve awareness and knowledge of Campylobacter, to advance food safety behaviors in domestic kitchens by improving attitudes, normative believes and perceived behavioral control, and to enhance control of Campylobacter in domestic kitchens by cleaning, sanitizing and cooking behaviors. Two delivery methods were used to target two groups of parents of young children over two consecutive summers. The result demonstrated that a) Campylobacter risk awareness was increased in both social media and traditional education groups; b) increase in understanding of Campylobacter control strategies was observed: cooking of ground meat and poultry, cross contamination from raw poultry to fresh produce, and preparation of turkey hot dogs; c) behavior changes were indicated: perceived behavioral control decreased in social media outreach group while the concern over the disease increased; and d) consumer trust in science or the government was increased after intervention. e. National symposium on Campylobacter intervention. A national symposium on intervention of Campylobacter in poultry was held on December 3, 2017 in Chicago in conjunction with the 98th annual Conference of Research Workers in Animal Diseases (http://crwad.org/). The symposium provided the newest information on innovative strategies to control Campylobacter in the food chain of poultry production.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Jones, L., X. Zeng, and J. Lin. 2013. Development of promising vaccination strategies to reduce Campylobacter colonization in chickens. 113rd General Meeting of American Society for Microbiology, May 18-21, Denver.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Jones, L., X. Zeng, and J. Lin. 2012. Development of novel vaccines for mitigation of Campylobacter in poultry. 93rd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 2-4, Chicago.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Issmat I. Kassem, M. Wolboldt, G. Rajashekara. Chemical treatments to reduce Campylobacter jejuni in chicken litter microcosms. The Annual Meeting of the Ohio Branch of American Society for Microbiology (OBASM), Ashland, OH. April 12-13, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Lin, J., L. Jones, X. Liu, and X. Zeng. Construction of novel DNA vaccine for mitigation of Campylobacter in chickens. P-2423. In Abstracts of the 114th General Meeting of American Society for Microbiology, Boston, MA. May 17-21, 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Sahin O, N. Pavlovic, A. O'Connor, D. Trampel, C. Logue, and Zhang, Q. A Longitudinal Study on Campylobacter Prevalence in Conventionally?Raised Commercial Broiler Flocks in the United States. P-1453. In Abstracts of 114th General Meeting of American Society for Microbiology, Boston, MA, May 17-21, 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Rajashekara G., Update on current food safety issues. Ohio Poultry Team and Tristate Poultry Veterinarian Meeting, Fort Recovery, OH, April 2014.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Sahin O, N. Pavlovic, A. O'Connor, C. Logue, and Zhang, Q. A Longitudinal Study on Campylobacter Prevalence in Conventionally?Raised Commercial Broiler Flocks in the United States. USDA National Institute of Food and Agriculture (NIFA) and Institute of Food Safety and Nutrition (IFSN) Project Director Meeting, July 24, 2015, Oregon, Portland.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Liu, X., X. Zeng, B. Gillespie, and J. Lin. Immune response and protective efficacy of intranasal vaccination of subunit vaccines for Campylobacter control in broilers. Annual Conference of Research Workers in Animal Disease. December 6-8, 2015, Chicago, IL.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: I.I. Kassem, O. O. Kehinde, A. Kumar, K. Chandrashekhar, R. Pina-Mimbela, G. Rajashekara. An evaluation of the impact of litter chemical amendments on reducing Campylobacter jejuni in broilers. Conference of research workers in animal disease (CRWAD), Chicago, IL. December 6- 8, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: I.I. Kassem, O. O. Kehinde, A. Kumar, K. Chandrashekhar, R. Pina-Mimbela, H. Huang, G. Rajashekara. Litter Chemical Amendments that Reduce pH and Moisture Contribute to the Control of Campylobacter jejuni in Broilers. The 115th General Meeting of the American Society for Microbiology, New Orleans, LA. May 30-June 2, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Logue CM, Barbieri NL, Cavender T, Baker A, O'Connor AM, and Zhang, Q. Campylobacter Associated with Poultry Processing  A View of Line Contamination. USDA National Institute of Food and Agriculture (NIFA) and Institute of Food Safety and Nutrition (IFSN) Project Director Meeting, July 30, 2016, St. Louis, Missouri.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Sahin O, Pavlovic N, OConnor A, Logue C, and Zhang Q. A longitudinal study on Campylobacter in commercial broiler flocks in the United States: Prevalence, genetic diversity, and associated risk factors. USDA National Institute of Food and Agriculture (NIFA) Project Directors Meeting, July 8, 2017, Tampa, FL. Oral presentation.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Sahin O, Looft T, Shen Z, Singh K, Zhang Q. Effect of the Intestinal Microbiota on Campylobacter jejuni Colonization in Broiler Chickens. Poster Presentation at ASM Microbe conference meeting, June 1-5, 2017, New Orleans, LA.
  • Type: Book Chapters Status: Published Year Published: 2016 Citation: I. I. Kassem, O. Kehinde, Y. A. Helmy, R. Pina-Mimbela, A. Kumar, K. Chandrashekhar, G. Rajashekara. 2016. Campylobacter in poultry: the conundrums of highly adaptable and ubiquitous foodborne pathogens. In: J. Mei Soon, L. J. Manning, C. A. Wallace (eds.), Foodborne Diseases: Case Studies of Outbreaks in the Agri-Food Industries. CRC Press, Taylor & Francis Group, USA. pp 79-112. ISBN: 978-1-4822-0827-6.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Sahin O, Yuan C, OConnor A, Logue C, Zhang Q. 2017. On-farm risk factors associated with Campylobacter prevalence in conventional broiler flocks in the United States. Poster presentation at CHRO Conference Meeting, September 10-14, 2017, Nantes, France.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Orhan Sahin and Qijing Zhang. Epidemiology and risk factors of Campylobacter in commercial broiler production. Oral presentation at National Symposium on Campylobacter Intervention*, December 3, 2017, held in conjunction with Conference of Research Workers in Animal Diseases Meeting, December 3-5, 2017, Chicago, IL.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Orhan Sahin and Qijing Zhang. Preharvest control strategies for Campylobacter in poultry. 2018. Campylobacter workshop: Managing the performance standards for poultry. Department of Poultry Science, University of Georgia, June 5-7, 2018, Athens, Georgia.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sahin O, I.I. Kassem, Z. Shen, J. Lin, G. Rajashekara, and Q. Zhang. 2015. Campylobacter in Poultry: Ecology and Potential Interventions. Avian Diseases, 59:185-200.
  • Type: Book Chapters Status: Published Year Published: 2016 Citation: Liu, X., I. Hanning, S. Diaz-Sanchez, and J. Lin. 2016. Food safety control on poultry farms: effective control of Campylobacter. Chapter 13. In Steven Ricke (ed), Achieving Sustainable Production of Poultry Meat. Burleigh Dodds Science Publishing, Cambridge, UK.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: I. Kassem, O. Kehinde, A. Kumar, and G. Rajashekara. 2017. Antimicrobial-resistant Campylobacter in organically and conventionally raised layer chickens. Foodborne Pathogens and Disease. 14(1):29-34.
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2018 Citation: Zhang Q, and Sahin O. 2017. Campylobacteriosis. Disease of Poultry, 14th Edition. Edited by David E. Swayne, Martine Boulianne, Larry R. McDougald, Venugopal Nair, Catherine M. Logue, and David L. Suarez. (This is an invited chapter and the manuscript is currently In Press).
  • Type: Websites Status: Published Year Published: 2017 Citation: Sahin O. 2017. Campylobacter in Poultry: An Elusive Pathogen. The Poultry Site: http://www.thepoultrysite.com/poultrynews/38164/campylobacter-in-poultry-an-elusive-pathogen/.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Kassem II, Helmy YA, Kathayat D, Candelero-Rueda RA, Kumar A, Deblais L, Huang HC, Sahin O, Zhang Q, Rajashekara G. 2017. Nonculturability Might Underestimate the Occurrence of Campylobacter in Broiler Litter. Foodborne Pathog Dis. 2017 Aug; 14(8):472-477. doi: 10.1089/fpd.2017.2279.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Kassem II, Candelero-Rueda RA, Esseili KA, Rajashekara G. 2017. Formate simultaneously reduces oxidase activity and enhances respiration in Campylobacter jejuni. Sci Rep. 2017 Jan 16; 7:40117. doi: 10.1038/srep40117.
  • Type: Journal Articles Status: Submitted Year Published: 2018 Citation: Liu, X., L. Jones, X. Zeng, J. Lin. 2018. Evaluation of in ovo vaccination of DNA vaccines for Campylobacter control in broiler chickens. Vaccine (Submitted).
  • Type: Journal Articles Status: Submitted Year Published: 2018 Citation: Adams, L.J., X. Zeng, J. Lin. 2018. Development and evaluation of two live Salmonella-vectored vaccines for Campylobacter control in broiler chickens. Foodborne Pathogens and Diseases (Submitted).
  • Type: Journal Articles Status: Other Year Published: 2018 Citation: Adams, L.J., X. Liu, L. Jones, Q. Zhong., X. Zeng, and J. Lin 2018. Evaluation of intranasal immunization with chitosan encapsulated subunit vaccines for Campylobacter control in broiler chickens. (In Preparation).
  • Type: Journal Articles Status: Other Year Published: 2018 Citation: Orhan Sahin, Nada Pavlovic, Yizhi Tang, Melda M. Ocal, Chaohui Yuan, Annette M. OConnor, Catherine M. Logue, Qijing Zhang. A longitudinal study on Campylobacter in commercial broiler flocks in the United States: Prevalence, genetic diversity, and associated risk factors (In Preparation).


Progress 07/15/16 to 07/14/17

Outputs
Target Audience:Poultry producers and processors, consumers, veterinarians, scientists, students. Changes/Problems:The meat safety training short course was originally planned to be held at Iowa State University. To increase the impact of the workshop, the course will now be taken place at the University of Georgia. The course will be organized by Dr. Manpreet Singh at Georgia in collaboration with Dr. Jim Dickson at ISU. Dr. Singh is a professor at the Department of Poultry Science and Extension, and is a well-known expert on food safety, poultry processing, and Campylobacter. He also has extensive connections with the poultry industry. What opportunities for training and professional development has the project provided?One undergraduate and one graduate students were trained under this project. How have the results been disseminated to communities of interest?Findings from this project have been presented at national conferences and published in peer-reviewed journals. The dedicated webpage on Campylobacter in poultry continues to help disseminate most relevant and current information on preharvest and postharvest food safety and control measures to poultry producers and processors, and consumers. Abstracts of 55 publications related to Campylobacter in poultry have been edited, added to the website, and linked to the original papers. What do you plan to do during the next reporting period to accomplish the goals?Based on the findings from the epidemiological/statistical analysis, major factors that are associated with on-farm Campylobacter prevalence (such as paw quality and percent burnt) will be evaluated using intervention studies. Data from the processing plant study will be analyzed to identify critical control points. All the research findings will be presented at relevant conferences and published in peer-reviewed journals. We will continue to update the website dedicated to Campylobacter in poultry to disseminate the relevant information to stakeholders. A meat safety training workshop has been planned for early Spring of 2018. Additionally, a National Campylobacter Symposium will be held on December 3, 2017 in Chicago, in conjunction with annual Conference of Research Workers in Animal Diseases. The symposium will also include a panel discussion on practical intervention of Campylobacter in poultry.

Impacts
What was accomplished under these goals? The on-farm risk factor assessment study to determine the factors associated with low prevalence/absence or high prevalence/presence of Campylobacter in the U.S. broiler flocks was completed. The hypothesis-generating analysis was done by univariate models using the Campylobacter prevalence data from 434 flocks raised in 53 different houses on 15 different farms (i.e., a longitudinal study covering 6-10 consecutive production cycles over a 2-year period) and 46 explanatory variables related to farm/house/flock-level specific standards, procedures, environmental conditions, and management practices. The analysis found that Campylobacter positivity/high prevalence was associated with increased feed conversion, increased adjusted prime cost, and decreased paw quality/increased percent hock burnt. Conversely, the variables of increasing average weight, increasing average daily gain, increasing feed conversion adjustment factor values, practice of litter treatment with different chemicals, increasing house temperature, increasing air litter temperature, the condition of air ammonia level (being less than 25 ppm) and litter amendment being used at proper rate were all associated with Campylobacter negativity/low prevalence. These findings identified several variables that may significantly influence the prevalence of Campylobacter on U.S. commercial broiler farms. Evaluation of these specific variables (e.g., paw quality/percent burnt), alone or in combination, under laboratory and/or commercial settings, is warranted in order to prove their impact and to develop and implement effective interventions for controlling Campylobacter on poultry farms Species identification and genetic diversity of the Campylobacter isolates from the cecal contents is completed. PCR identified most of the isolates as C. jejuni (88.6%; 2364 of 2666) and almost all of the rest as C. coli (11%; 294 of 2666). The remaining 8 isolates were Campylobacter species other than C. jejuni or C. coli. The vast majority of C. coli isolates were from only a few farms while some farms did not have this species at all during the entire sampling period. Genotyping of C. jejuni isolates (included 346 representative isolates) via pulsed-field gel electrophoresis (PFGE) showed the presence of 35 clusters at a 90% similarity level. Multilocus sequence typing (MLST) of representative isolates (1-2 isolates/PFGE type) found 8 known sequence types and 15 novel STs. The most common ST types are ST939, ST467, ST353, and ST222. The result indicated that the overall genetic diversity within a farm was limited, mostly only a couple of genotypes, and that some genotypes persisted through all production cycles. In addition, the genotypes were mostly farm-specific even though some of them were observed on multiple farms. These findings indicate either survival of certain Campylobacter genotypes in the poultry environment for a long period or/and presence of a common source(s) in the farm environment for flock infection. For the extension component, the dedicated Campylobacter website continues to disseminate the new and relevant information to the stakeholders. The website is continuously being updated with the abstracts of the most recent peer-reviewed journal publications with links to the full articles. Additionally, the web site is updated with information on an upcoming national symposium on Campylobacter intervention in poultry.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Sahin O, Pavlovic N, OConnor A, Logue C, and Zhang Q. A longitudinal study on Campylobacter in commercial broiler flocks in the United States: Prevalence, genetic diversity, and associated risk factors. USDA National Institute of Food and Agriculture (NIFA) Project Directors Meeting, July 8, 2017, Tampa, FL. Oral presentation
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Sahin O, Looft T, Shen Z, Singh K, Zhang Q. Effect of the Intestinal Microbiota on Campylobacter jejuni Colonization in Broiler Chickens. Poster Presentation at ASM Microbe conference meeting, June 1-5, 2017, New Orleans, LA.
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2017 Citation: Zhang Q, and Sahin O. 2017. Campylobacteriosis. Disease of Poultry, 14th Edition. Edited by David E. Swayne, Martine Boulianne, Larry R. McDougald, Venugopal Nair, Catherine M. Logue, and David L. Suarez.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Kassem II, Helmy YA, Kathayat D, Candelero-Rueda RA, Kumar A, Deblais L, Huang HC, Sahin O, Zhang Q, Rajashekara G. 2017. Nonculturability Might Underestimate the Occurrence of Campylobacter in Broiler Litter. Foodborne Pathog Dis. 2017 Aug; 14(8):472-477. doi: 10.1089/fpd.2017.2279.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Kassem II, Candelero-Rueda RA, Esseili KA, Rajashekara G. 2017. Formate simultaneously reduces oxidase activity and enhances respiration in Campylobacter jejuni. Sci Rep. 2017 Jan 16; 7:40117. doi: 10.1038/srep40117.


Progress 07/15/15 to 07/14/16

Outputs
Target Audience:Poultry producers and processors, consumers, veterinarians, students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One undergraduate and one graduate students were trained under this project. How have the results been disseminated to communities of interest?Findings from this project have been presented at national conferences and published in peer-reviewed journals. The dedicated webpage on Campylobacter in poultry continues to help disseminate most relevant and current information on preharvest and postharvest food safety and control measures to poultry producers and processors, and consumers. Abstracts of 50 scientific publications have been edited, added to the website, and linked to the complete papers so far. Social media sites including a website, Facebook page, Twitter Account, YouTube and Pinterest have been constructed for consumer education on safe handling of chicken and turkey meat. Developed curriculums were distributed to the participants (parenting adults of children) as six lessons via email and text messages with embedded links to the OSU website and outside sites such as FDA, USDA and CDC. Live links to SurveyMonkey online surveys were prepared and embedded in emails for each evaluation instrument.These emails were delivered pre/post and at the end of each weekly lesson. What do you plan to do during the next reporting period to accomplish the goals?Based on the findings from the epidemiological/statistical analysis, major factors that are associated with on-farm Campylobacter prevalence will be evaluated using intervention studies. Data from the processing plant study will be analyzed to identify critical control points; the most important variables from this analysis will be tested for their effect on Campylobacter carriage on carcasses using a pilot plant study. All the research findings will be continued to be presented at relevant conferences and published in peer-reviewed journals. We will continue to update the website dedicated to Campylobacter in poultry to disseminate the relevant information to stakeholders. We will hold the meat safety training short course at the Meat Laboratory at ISU, and National Campylobacter Symposium. For consumer education component, data analysis will commence after all data are collected. Additionally, EFNEP data is being currently analyzed for the study comparison group. Data will be used to prepare manuscripts for review and publication.

Impacts
What was accomplished under these goals? To determine the factors contributing to low or high prevalence of Campylobacter on poultry farms, detailed information on the production and management characteristics of the study broiler farms was collected. The data covered flock-, house-and farm-level parameters and included information from 434 flocks raised in 53 different houses on 15 different farms (i.e., a longitudinal study covering 6-10 consecutive production cycles over a 2-year period). Analysis of the data revealed a number of factors that were associated with high/presence- or low/absence level of Campylobacter prevalence in the broiler flocks/houses/farms. Among the variables that were found to be associated with Campylobacter positivity/high prevalence are increased feed conversion, increased adjusted prime cost, and decreased paw quality/increased percent burnt (all flock-level data). Among the variables that were found to be associated with Campylobacter negativity/low prevalence are increasing average weight, increasing average daily gain, increasing feed conversion adjustment factor values, practice of litter treatment with different chemicals, increasing house temperature, increasing air litter temperature, the condition of air ammonia level being less than 25 ppm and litter amendment being used at proper rate, and perfect placement variable water being 15-25 ml/min. Species identification of the Campylobacter isolates from the cecal contents is completed. PCR identified most of the isolates as C. jejuni (88.6%; 2364 of 2666) and almost all of the rest as C. coli (11%; 294 of 2666). The remaining 8 isolates were Campylobacter species other than C. jejuni or C. coli. Genotyping of C. jejuni isolates via pulsed-field gel electrophoresis (PFGE) is finished (included 346 isolates representing the isolations from all positive-farms/houses/flocks on every production cycle). Data indicates the existence of limited number of genotypes and persistence of a few clones over multiple cycles on the same farm. A representative set of isolates (1 isolate/PFGE type) are further analyzed using MLST. Processing plant study has been finished. Carcass-wash samples (n= 2030) were collected from 9 different sites along the process line at 9 commercial slaughterhouses. Of these samples, 580 (28.6%) were positive for Campylobacter spp., with the majority (525; 90.5%) yielding C. jejuni The majority of samples that were positive for Campylobacter occurred early in the process line, and detection at the later stages was extremely low. Different plants examined had different prevalence levels and could be categorized into high-medium and low prevalence plants. These data suggest that interventions at the latter stages of processing have a significant impact in reducing the overall final carcass contamination. Litter treatment study has now been completed. In order to investigate the efficacy of on-farm practices to control litter as a source of Campylobacter infection within and between flocks, experiments were carried out. We selected 4 broiler farms (2 with low-prevalence of Campylobacter and 2 with high prevalence of Campylobacter). All these farms used hot air treatment of litter before placing new birds. We sampled litter (n = 384) and ceca (n = 192) from 6 houses on each farm for two growth cycles. The samples were collected during downtime after treatment and at the end of the growth cycle. The samples were screened for Campylobacter, and pH and moisture of the litter were also monitored throughout the experiment. The hot air treatment of the litter increased the alkalinity of the litter (although it was not always significant and varied based on farm), and reduced moisture of the litter. Nine litters samples that were collected during growth cycle from different farms were positive for Campylobacter. The hot air treatment did not result in reducing the Campylobacter in the flocks in the high prevalence farms. In one of the low prevalence farms, cecal and litter samples remained Campylobacter negative pre- and post- hot air treatment, while in the other low prevalence farm cecal samples became Campylobacter positive post-treatment. These findings indicate that the hot air treatment did not show consistent impact on Campylobacter colonization of the flocks. For the extension component, the dedicated Campylobacter website continues to disseminate the new and relevant information to the stakeholders. The website is continuously being updated with the abstracts of the most recent peer-reviewed journal publications with links to the full articles. For consumer education, Phase 2 of data collection was completed. Six Ohio County Educators volunteered to recruit participants in Phase 2. As of this writing, 213 participants were recruited to the Social Media Online Education (SMOE) group and 128 to the Traditional Online Education (TOE) group for a total of 341 for the study. All data will be downloaded for final statistical analysis.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Logue CM, Barbieri NL, Cavender T, Baker A, O'Connor AM, and Zhang, Q. Campylobacter Associated with Poultry Processing  A View of Line Contamination. USDA National Institute of Food and Agriculture (NIFA) and Institute of Food Safety and Nutrition (IFSN) Project Director Meeting, July 30, 2016, St. Louis, Missouri.
  • Type: Book Chapters Status: Accepted Year Published: 2016 Citation: Liu, X., I. Hanning, S. Diaz-Sanchez, and J. Lin. 2016. Food safety control on poultry farms: effective control of Campylobacter. Chapter 11. In Steven Ricke (ed), Achieving Sustainable Production of Poultry Meat. Burleigh Dodds Science Publishing, Cambridge, UK (In Press)
  • Type: Book Chapters Status: Published Year Published: 2016 Citation: I. I. Kassem, O. Kehinde, Y. A. Helmy, R. Pina-Mimbela, A. Kumar, K. Chandrashekhar, G. Rajashekara. 2016. Campylobacter in poultry: the conundrums of highly adaptable and ubiquitous foodborne pathogens. In: J. Mei Soon, L. J. Manning, C. A. Wallace (eds.), Foodborne Diseases: Case Studies of Outbreaks in the Agri-Food Industries. CRC Press, Taylor & Francis Group, USA. pp 79-112. ISBN: 978-1-4822-0827-6.
  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: I. Kassem, O. Kehinde, A. Kumar, and G. Rajashekara. 2016. Antimicrobial-resistant Campylobacter in organically and conventionally raised layer chickens. Foodborne Pathogens and Disease. In press.


Progress 07/15/14 to 07/14/15

Outputs
Target Audience:Poultry producers and processors, consumers, veterinarians, students Changes/Problems:None. What opportunities for training and professional development has the project provided?One undergraduate and one graduate students were trained under this project. How have the results been disseminated to communities of interest?Findings from this project have been presented at national conferences and published in peer-reviewed journals. The dedicated webpage on Campylobacter in poultry continues to help disseminate most relevant and current information on preharvest and postharvest food safety and control measures to poultry producers and processors, and consumers. Abstracts of 40 scientific publications have been edited, added to the website, and linked to the complete papers so far. Social media sites including a website, Facebook page, Twitter Account, YouTube and Pinterest have been constructed for consumer education on safe handling of chicken and turkey meat. Developed curriculums were distributed to the participants (parenting adults of children) as six lessons via email and text messages with embedded links to the OSU website and outside sites such as FDA, USDA and CDC. Live links to SurveyMonkey online surveys were prepared and embedded in emails for each evaluation instrument.These emails were delivered pre/post and at the end of each weekly lesson. What do you plan to do during the next reporting period to accomplish the goals?The low- and high-prevalence status of the study farms will be continuously monitored by Campylobacter testing. Major efforts will be made on identifying factors associated with Campylobacter presence/absence at the preharvest level. Intervention studies based on the findings will be initiated. Data from the processing plant study will be analyzed to identify critical control points. We will finish the final phase of the litter treatment study in which the most effective chemical treatment scheme will be used to reduce Campylobacter prevalence in a commercial farm setting. All the research findings will be continued to be presented at relevant conferences and published in peer-reviewed journals. We will continue to update the website dedicated to Campylobacter in poultry to disseminate the relevant information to stakeholders. We will hold the first meat safety training short course at the Meat Laboratory at ISU; the course will emphasize Campylobacter in poultry for industry and government agencies. For consumer education component, recruitment of participants will be finished soon (Phase 1, Data collection), and another recruitment will begin again (Phase 2, Data Collection). The last group of subjects beginning their 6 lesson series will complete their post-test and active data collection will then be completed. Data analysis will commence after Phase 1 Data Collection, and additional EFNEP data will be analyzed for the study comparison group. Data will be used to compile abstracts for national presentations and preliminary peer-review papers will be prepared and submitted for publication.

Impacts
What was accomplished under these goals? The longitudinal study on 15 commercial broiler farms (as part of the same integrated production system) was completed (lasted about 18 months) and identified farms/houses with consistently low- or high- Campylobacter prevalence level. Continuous testing of some of the target farms in has been performed to generate more specific data in line with the project goals. Visits were conducted to the low- and high- prevalence farms to collect detailed farm/house/flock level information, which will soon be analyzed to determine the factors associated with low or high prevalence. Species identification of the Campylobacter isolates from the cecal contents is nearly finished. PCR identified most of the isolates as C. jejuni (88.6%; 2364 of 2666) and almost all of the rest as C. coli (11%; 294 of 2666). Genotyping of C. jejuni isolates via pulsed-field gel electrophoresis (PFGE) is currently undergoing, and the preliminary data indicates the existence of limited number of genotypes and persistence of a few clones over multiple cycles on the same farm. Processing plant study has been initiated and carcass-wash samples (n= 2030) were collected from 9 different sites along the process line at 9 slaughterhouses. Of these samples, 580 (28.6%) were positive for Campylobacter spp., with the majority (525; 90.5%) yielding C. jejuni and the rest (55; 9.4%) having C. coli. The Campylobacter counts ranged from 0.7 to > 6 log10 cfu/ml. The majority of samples that were positive for Campylobacter occurred early in the process line (i.e. pre and post scald, and rehang stage), and detection at the later stages was minimal. Several trails were conducted with the immunization study. In one chicken trial, six groups of broilers (9-14 birds per group) received different formulation of DNA and subunit protein vaccines in ovo with several modifications in order to prevent degradation of DNA in amniotic fluid; however, the results indicated that the new formulation of DNA vaccines as well as the subunit protein vaccines did not trigger a strong immune response and did not protect against C. jejuni colonization. In another chicken trial, we observed that intranasal vaccination with chitosan encapsulated protein subunit vaccines significantly elicited serum IgG and mucosal IgA response although DNA vaccine given in a similar way did not induce any significant immune response. Litter treatment study was continued in the past year. In a small pen trial that was conducted to determine the effect of treating litter with ammonium sulfate (AS) + magnesium sulfate (MS) or AS +MS + sodium bisulfate (SOB), the bacterial communities in the litter and chickens were evaluated. For this purpose, 18 cecal samples and 6 litter samples were strategically selected to conduct metagenomic analysis (16S rRNA gene sequencing). Results indicated that Bacilli decreased from 28% in control litter to 2.8% in treated litter. There was no similar effect on the Bacilli in the cecal samples. Likewise, the Erysipelotrichaceae class decreased from 2.5% in control litter to 0.1% in treated litter. A similar effect was observed in the cecal samples (6% in control vs. 0.8% in treated samples). Also, it was found that the order Campylobacterales decreased from an average of 2% in untreated ceca to 0.2% in treated ceca. For the extension component, the dedicated Campylobacter website continues to disseminate the new and relevant information to the stakeholders. The website is continuously being updated with the abstracts of the most recent peer-reviewed journal publications with links to the full articles. For consumer education, social media sites have been constructed, including a website, Facebook page, Twitter Account, YouTube and Pinterest. Two curriculums were developed based on principles of consumer safe food handling of chicken and turkey during the summers months, and that were aimed at parenting adults of children, ages 1 to 9 (the at-risk age group for campylobacteriosis). One curriculum, titled the Traditional Outreach Education (TOE) group, was distributed to participants as six lessons primarily sent via email, with some text messages. The other curriculum, titled the Social Media Outreach Education (SMOE) group, was distributed to participants as six lessons primarily sent via text messages, with some email messages. Also, evaluation instruments were completed; pre-study survey, post-study survey, and weekly participation inventories to document activities completed during each of the six lessons.Live links to SurveyMonkey online surveys were prepared and embedded in emails for each evaluation instrument.Recruitment materials and consent forms were prepared, and recruitment began the 3rd week of June 2015 with the first consent signed and subject enrolled by the 4the week of June 2015. So far, 171 participants were recruited to the SMOE group and 96 to the TOE group. Twelve county educators volunteered to recruit participants.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2015 Citation: Sahin O, I.I. Kassem, Z. Shen, J. Lin, G. Rajashekara, and Q. Zhang. 2015. Campylobacter in Poultry: Ecology and Potential Interventions. Avian Diseases, 59:185-200.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: Sahin O, N. Pavlovic, A. O'Connor, C. Logue, and Zhang, Q. A Longitudinal Study on Campylobacter Prevalence in Conventionally?Raised Commercial Broiler Flocks in the United States. USDA National Institute of Food and Agriculture (NIFA) and Institute of Food Safety and Nutrition (IFSN) Project Director Meeting, July 24, 2015, Oregon, Portland
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: Liu, X., X. Zeng, B. Gillespie, and J. Lin. Immune response and protective efficacy of intranasal vaccination of subunit vaccines for Campylobacter control in broilers. Annual Conference of Research Workers in Animal Disease. December 6-8, 2015, Chicago, IL
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: I.I. Kassem, O. O. Kehinde, A. Kumar, K. Chandrashekhar, R. Pina-Mimbela, G. Rajashekara. An evaluation of the impact of litter chemical amendments on reducing Campylobacter jejuni in broilers. Conference of research workers in animal disease (CRWAD), Chicago, IL. December 6- 8, 2015
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: I.I. Kassem, O. O. Kehinde, A. Kumar, K. Chandrashekhar, R. Pina-Mimbela, H. Huang, G. Rajashekara. Litter Chemical Amendments that Reduce pH and Moisture Contribute to the Control of Campylobacter jejuni in Broilers. The 115th General Meeting of the American Society for Microbiology, New Orleans, LA. May 30-June 2, 2015
  • Type: Journal Articles Status: Submitted Year Published: 2015 Citation: Liu, X., X. Zeng, L. Jones, J. Lin. 2015. Introduction of the Kozak consensus sequence in DNA vaccine enhanced the expression of Campylobacter genes in mammalian cells. Submitted to Synthetic and Systems Biotechnology


Progress 07/15/13 to 07/14/14

Outputs
Target Audience: Poultry producers and processors, consumers, veterinarians, students Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Four undergraduate students were trained under this project. How have the results been disseminated to communities of interest? The dedicated Campylobacter webpage launched to help disseminate relevant information to poultry producers and processors, and consumers has been continually updated with new information to promote food safety. Currently, 30 abstracts of scientific papers have been edited, added to the website, and linked to the complete papers. The “Consumer” webpage provides food safety information that will help consumers avoid the foodborne disease caused by Campylobacter. Tips for preventing cross contamination of raw food products and elimination of bacterial pathogens by properly cooking poultry meat prior to human consumption are given. The “Consumer” webpage has 3 sections which communicate a) clinical signs associated with human food poisoning caused by Campylobacter, b) potential sources of contamination, and c) methods to prevent food poisoning, including the four food safety steps of USDA's Food Safe Families campaign. Additional information on safe handling of chicken meat is presented as well. The “Producer” webpage describes Campylobacter colonization of clinically healthy poultry and lists potential sources of infection for broiler chickens and turkeys. The most current information available concerning pre-harvest control strategies to reduce the level of Campylobacter colonization in live birds is outlined. The “Processor” webpage includes information concerning Campylobacter contamination of broiler and turkey carcasses and potential sources of Campylobacter carcass contamination. In addition, postharvest methods to reduce numbers of Campylobacter on poultry carcasses and contamination of poultry meat products are presented. What do you plan to do during the next reporting period to accomplish the goals? We will finish collecting samples from the majority of currently tested broiler farms, but focus our efforts on more targeted farms for generation of more specific data in line with the project goals, continue species identification and characterization of the isolates. We will also use more specific and detailed farm/house/flock level information and continue analyzing them in relation to the prevalence data to determine factors that may affect Campylobacter presence/absence in broiler flocks. Processing plant survey will be initiated in the third year of the project. We will first identify the study processing plants in order to determine the processing steps that could be managed for reducing Campylobacter during slaughter. We will determine the prevalence and concentration of Campylobacter before and after six processing-points on the poultry processing line (slaughter, scald, defeathering, evisceration, wash, and chill). We will perform multiple chicken studies to evaluate the efficacy of different vaccines that have been successfully prepared in this period (in ovo DNA vaccines and intranasal subunit vaccines) for prevention of Campylobacter colonization in chickens. We will continue the litter management experiments. The goals are to 1) analyze microbiota of chicken ceca that were negative for Campylobacter and compare that with chickens that were positive for Campylobacter following litter treatment by next generation sequencing, 2) communicate our results to the stakeholder through our extension and outreach activities and further we will disseminate our results to the scientific community through publications in peer reviewed journals, 3) apply the best chemical treatment from the above studies on a large scale farm operation setting, and 4) evaluate new chemicals for C. jejuni control. For example, our preliminary studies have indicated that that at optimal concentration, formic acid and succinic acid (both are weak acids) can inhibit Campylobacter growth and biofilm formation. The nature and impact of these acids makes them ideal for use as water additives. Therefore, these acids will be used in tandem to enhance the impact of the litter treatment studies. We will continue to enrich the website dedicated to Campylobacter in poultry that has been established to disseminate relevant information. More scientific abstracts will be added. Links to webpages for project co-directors, to USDA’s 4 food safety steps, and to other locations will be added on the webpage. In addition, we will begin to develop written materials to disseminate new information generated by research team members that will eventually be submitted to journals whose readership includes consumers of poultry products, broiler and turkey producers, and broiler and turkey processors. With respect to consumer education component, the web manager will complete the web site in the 3rd year and post live by July 2014. Once basic construction and design is completed, all educational materials to be used to implement the research study will be finalized. Key personnel located in Ohio County Extension Offices will be recruited to complete local subject recruiting and research study initiation. All Key Personnel and all educational materials seen by subjects will be submitted for human subjects review and approval. The intervention research study will begin in May 2015.Data from the EFNEP program for the baseline comparison group has been received and will be statistically analyzed this year.

Impacts
What was accomplished under these goals? Prevalence of Campylobacter at the flock (45.2%), house (77.3%), and farm (93.3%) levels were obtained. Within-flock prevalence rates were 94.3% by cecal contents (ranging from 6.6 to 100%) and 79.3% by boot swabs (ranging from 33.3 to 100%) in the positive flocks. The mean level of Campylobacter numbers in positive flocks was 5.3 (ranging from 0.8 to 7.7) log10 CFU/g feces by cecal contents and 3.4 (ranging from 0.7 to 6.6) log10 CFU/g feces by boot swabs. Some farms consistently lacked Campylobacter, while some had always have it. Species identification of Campylobacter isolates is continuously being performed using PCR and other methods, which indicated vast majority as being C. jejuni. Vaccines with various formulations (including DNA and subunit proteins) have been developed successfully and some vaccination regimens have been evaluated for their potency against Campylobacter colonization in chickens. Effect of various litter treatments were evaluated in separate trals. In trial I, orally challenged broilers (with C. jejuni) were placed in rooms with different litter treatments. In trial 2, uninfected birds were placed into rooms containing C. jejuni-contaminated litter that was treated with different chemicals. In trial 3, broiler chickens (including uninfected and C. jejuni-inoculated seeder birds) were placed into rooms containing litter treated with different chemicals. In all three phases of our study, the pH and moisture of the litter significantly decreased when the treatment were applied at the correct dosage at the beginning of the experiment. This effect lasted till the end of 6 week period. The best treatment was found to be the inclusion of all 3 chemicals together. This treatment resulted in the most consistent and stable reduction of litter’s pH and moisture. Furthermore, the combination of the 3 chemicals was effective in significantly reducing the colonization of chickens and litter with Campylobacter in trial II. None of the chemical amendments affected the performance of the chickens in terms of weight gain, feed conversion, and mortality. Treatment of the litter did not significantly reduce the load of C. jejuni in experimentally inoculated chickens or in litter (trial I); however, it significantly protected chickens from being colonized with C. jejuni, and the litter remained free of C. jejuni (trial II), which was especially evident when using the AS + SOB + MS treatment. Finally, even though treatment of the litter did not prevent transmission of C. jejuni from orally inoculated seeder chickens to un-inoculated chickens, the litter was Campylobacter-free under this scenario (trial III). This Campylobacter website disseminates current and relevant information to consumers of poultry products, poultry producers, and poultry processors. Google Analytics has been used to track usage of the web site. Peer-reviewed papers published in refereed scientific journals are the source of all abstracts appearing on the Campylobacter website. Abstracts are added to the “Campylobacter News” section of the website on a regular schedule. As new abstracts are added, older abstracts are removed from the homepage and placed in an archive which is accessible to viewers. Each abstract is linked to a pdf file containing the complete paper in a refereed journal. Abstracts are edited to make them understandable to individuals without a microbiology background. News items are brief descriptions which enable the reader to determine if they wish to view the complete paper, as opposed to complete abstracts of the original paper. Currently, 30 abstracts of scientific papers have been edited, added to the website, and linked to the complete papers. For consumer education, baseline comparison group data has been requested from the Ohio EFNEP Program. In addition, the preliminary pre/post survey has been developed. The survey instruments to be used in the present study is presently being revised to take advantage of lessons learned from the previous studies, the topic of Campylobacter control in consumer kitchens, summer food selection, preparation and storage, and behavioral beliefs and belief evaluations of the parenting target population of this project. Once completed, the project will generate important information that can be utilized to control Campylobacter in poultry, which is a significant source of foodborne illnesses. The outcomes will improve food safety at multiple levels of production and consumption, and will benefit the U.S. poultry industry by helping them meet the new performance standards set by USDA-FSIS.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2014 Citation: Rajashekara G., Update on current food safety issues. Ohio Poultry Team and Tristate Poultry Veterinarian Meeting, Fort Recovery, OH, April 2014.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2014 Citation: Lin, J., L. Jones, X. Liu, and X. Zeng. Construction of novel DNA vaccine for mitigation of Campylobacter in chickens. P-2423. In Abstracts of the 114th General Meeting of American Society for Microbiology, Boston, MA. May 17-21, 2014.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2014 Citation: Sahin O, N. Pavlovic, A. O'Connor, D. Trampel, C. Logue, and Zhang, Q. A Longitudinal Study on Campylobacter Prevalence in Conventionally?Raised Commercial Broiler Flocks in the United States. P-1453. In Abstracts of 114th General Meeting of American Society for Microbiology, Boston, MA, May 17-21, 2014.


Progress 07/15/12 to 07/14/13

Outputs
Target Audience: The target audience includes poultry producers and processors, consumers, veterinarians, researchers, and graduate students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Three undergraduate students were trained under this project. How have the results been disseminated to communities of interest? The Campylobacter website include 1) a unique banner, 2) rotating photographs, 3) current Campylobacter news items, and 4) links to webpages which provide information about this project and information for producers, 6) processors, and 7) consumers. There is a link from each item to the “Campylobacter News” location to the complete paper in a journal. The news items are to be understandable to consumers without a technical background. Peer-reviewed papers published in refereed scientific journals are the source of news items appearing on the webpage. On the Producer Webpage, Campylobacter colonization of clinically normal poultry, potential sources of infection, and pre-harvest control strategies are described. On the Processor Webpage, information includes Campylobacter on poultry carcasses, source of Campylobacter carcass contamination, and postharvest control of Campylobacter on poultry carcasses. Consumer webpage covers food safety information that will help consumers avoid foodborne disease caused by Campylobacter. What do you plan to do during the next reporting period to accomplish the goals? We will continue collecting samples from broiler farms, culturing them for Campylobacter, conduct species identification, and initiate molecular typing of the isolates. We will also start collecting farm/house/flock specific data and analyze them in relation to the culture results to determine factors that affect Campylobacter prevalence. We will perform multiple chicken studies to evaluate the various vaccination regimens (in ovo DNA vaccines, intranasal subunit vaccines, and oral attenuated Salmonella-vectored live vaccines) for prevention of Campylobacter colonization in chickens. We will undertake experiments to finalize the last remaining stages of the litter microcosm studies as proposed. We will test if our chemical amendments reduce litter and chicken contamination with C. jejuni. For this purpose, we will initiate the second part of our proposed studies. Specifically, we will select the chemical amendments that gave the best result in the microcosms trials. These amendments will then be applied to contained pens housing live chickens (experimentally infected or not infected with C. jejuni). During the second year, we will continue to develop the website. Links to other locations will be added. Information for processors will be expanded. We will attempt to provide videos for each of the 4 food safety steps recommended by USDA. In addition, we will provide additional means of information delivery to stakeholders, including exploring the possibility of using social media. With respect to consumer education component, we will hire a web manager to complete web database interface; modify web (foodsafety.osu.edu) with food safety information for control of Campylobacter and the target population and link the local web site with the overall project web site.

Impacts
What was accomplished under these goals? Prevalence of Campylobacter was 51.2% at flock level, 67.9% at house level, and 80% at farm level. Within-flock prevalence rates were found to be 93% by cecal contents (ranging from 0 to 100%) and 79.1% by boot swabs (ranging from 0 to 100%). Some farms were consistently lack of Campylobacter. The mean level of Campylobacter numbers in positive flocks was 5.4 (ranging from 0.2 to 7.7) log10 CFU/g feces by cecal contents and 3.6 (ranging from 0.7 to 6.6) log10 CFU/g feces by boot swabs. Species identification of Campylobacter isolates are currently being performed using PCR and other methods. For vaccination, various constructs were successfully made. The first chicken trial (80 chickens with four groups) is being performed to determine the immune response and efficacy of the Salmonella-vectored vaccines. For litter treatments, we have tested the impact of sodium bisulfate, ammonium sulfate and magnesium sulfate, individually and in combination, as potential chemical amendments to reduce the pH and moisture of broiler litter and C. jejuni contamination. Ammonium sulfate and sodium bisulfate had no significant effect on reducing the litter moisture; however, the addition of magnesium sulfate to the litter reduced the moisture by about 50%. Both ammonium sulfate and sodium bisulfate, individually and in combination, reduced the numbers of C. jejuni spiked into the litter microcosms. Depending on the treatment, significant reduction (about 2 log) in C. jejuni numbers were observed as early as 2 h post-treatment. In all treatments, C. jejuni was undetectable in the litter after 24 h of incubation. The Campylobacter website include 1) a unique banner, 2) rotating photographs, 3) current Campylobacter news items, and 4) links to webpages which provide information about this project and information for producers, 6) processors, and 7) consumers. There is a link from each item to the “Campylobacter News” location to the complete paper in a journal. The news items are to be understandable to consumers without a technical background. Peer-reviewed papers published in refereed scientific journals are the source of news items appearing on the webpage. On the Producer Webpage, Campylobacter colonization of clinically normal poultry, potential sources of infection, and pre-harvest control strategies are described. On the Processor Webpage, information includes Campylobacter on poultry carcasses, source of Campylobacter carcass contamination, and postharvest control of Campylobacter on poultry carcasses. Consumer webpage covers food safety information that will help consumers avoid foodborne disease caused by Campylobacter. It is organized in sections as Campylobacter and Food Poisoning, Sources of Campylobacter Food Poisoning, and Prevention of Campylobacter Food Poisoning (includes the four food safety steps of USDA's Food Safe Families campaign and additional information on safe handling of chicken meat). Information on the website will help producers and processors reduce the level of Campylobacter contamination on poultry products and will help consumers to safely prepare poultry meat for human consumption. The progress during the first year has ensured a smooth transition to the second year. Once the project is completed, the project will generate important information that can be utilized to control Campylobacter in poultry, which is a significant source of foodborne illnesses. The outcomes will improve food safety at multiple levels of production and consumption, and will benefit the U.S. poultry industry by helping them meet the new performance standards set by USDA-FSIS.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Jones, L., X. Zeng, and J. Lin. 2013. Development of promising vaccination strategies to reduce Campylobacter colonization in chickens. 113rd General Meeting of American Society for Microbiology, May 18-21, Denver.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Jones, L., X. Zeng, and J. Lin. 2012. Development of novel vaccines for mitigation of Campylobacter in poultry. 93rd Annual Meeting of Conference of Research Workers in Animal Diseases. Dec 2-4, Chicago.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Issmat I. Kassem, M. Wolboldt, G. Rajashekara. Chemical treatments to reduce Campylobacter jejuni in chicken litter microcosms. The Annual Meeting of the Ohio Branch of American Society for Microbiology (OBASM), Ashland, OH. April 12-13, 2013.