Source: UNIVERSITY OF GEORGIA submitted to NRP
PROCESSING ANTIMICROBIALS SELECT FOR PROCESSING TOLERANT CAMPYLOBACTER JEJUNI: IMPLICATIONS TO HEALTH AND POTENTIAL INTERVENTIONS.
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1030705
Grant No.
2023-67017-40016
Cumulative Award Amt.
$279,663.00
Proposal No.
2022-09038
Multistate No.
(N/A)
Project Start Date
Jul 1, 2023
Project End Date
Jun 30, 2025
Grant Year
2023
Program Code
[A1332]- Food Safety and Defense
Recipient Organization
UNIVERSITY OF GEORGIA
200 D.W. BROOKS DR
ATHENS,GA 30602-5016
Performing Department
(N/A)
Non Technical Summary
Campylobacter jejuni is a leading cause of foodborne gastroenteritis in humans. Contaminated poultry products are a significant source of C. jejuni. It is well-established that C. jejuni can con­taminate a high percentage of poultry carcasses at the end of processing.The latter can be due to several reasons; including the potential existence of certain C. jejuni strains that can tolerate commonly used antimicrobials. Here, we hypothesize that certain C. jejuni strains are tolerant to commonly used processing antimicrobials and that these strains, dubbed process tolerant C. jejuni, might be related to human disease. Therefore, the overall objective of this project isto identify C. jejuni that are tolerant to antimicrobials commonly used during poultry processing and to investigate an approach to control these problematic strains. Controlling C. jejuni is critical to improve the safety of poultry products and reduce adverse economic and public health consequences. This project serves the purpose of: 1) reducing problematic C. jejuni strains that threaten the public, and 2) providing the poultry industry with enhanced controls for safer products.
Animal Health Component
90%
Research Effort Categories
Basic
10%
Applied
90%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
71232601100100%
Goals / Objectives
The overall objective is to control Campylobacteron poutry carcasses in order to enhance the safety of poutry meat and enhance public health.Despite advances in pathogen control during poultry processing, it is established that Campylobacterjejuni, a leading foodborne pathogen,can con­taminate a high percentage of poultry carcasses at the end of processing. Although antimicrobials are deployed to reduce Campylobacter numbers during processing, it has been shown that the antimicrobials might show lowered effectiveness due to multiple factors, including carcass propertiesand/ or the high level of contamination associated with the large number of poultry in processing. However, an important factor that might have been overlooked is the tolerance of certain Campylobacter strains to antimicrobials and other controls used during processing. Despite the importance of these observations, there is limited data on the existence of potentially process tolerant C. jejuni and their link to human disease.Here, it ishypothesize that certain C. jejuni strains are tolerant to commonly used processing antimicrobials and that these strains, dubbed process tolerant C. jejuni, might be clinically relevant. Therefore, the major aim of this proposal is to identify C. jejuni that are tolerant to antimicrobials commonly used during poultry processing and propose an approach to control them. This will be achieved via the following objectives:Objective 1: Isolation and typing of C. jejuni from carcass samples in poultry plants.Objective 2: Assess the tolerance of C. jejuni to process antimicrobials (chlorine, peracetic acid, cetylpyridinium chloride, acidified sodium chlorite, organic acids: lactic acid).Objective 3: Control of process tolerant C. jejuni on artificially contaminated chicken carcasses.This proposal addresses the USDA-NIFA Program Area Priority Code, A1332. We are tackling an emerging food safety concern for both consumers and the industry. The data generated from the proposed research has the potential to be rapidly scaled up to enhance the safety of an essential protein source, poultry meat, during processing.
Project Methods
The FSIS-USDA targetsCampylobacteron carcasses in the post-chill area. Therefore, we will focus the analysis on post-chill carcasses to isolateCampylobacter. The isolates will be typed using Whole Genome Sequencing (WGS) to identify their properties, the genotypes that are process-tolerant, and their relationship to human-associated isolates. We will test the tolerance of these isolates to the most commonly used processing antimicrobials, specifically chlorine, peracetic acid, cetylpyridinium chloride, acidified sodium chlorite, and organic acids: lactic acid. Therefore, the methods used in this project include standard microbiological assays for the isolation and identification ofCampylobacter, evaluation of MICs, and the effectiveness of the antimicrobials. We will also deploy molecular methods such as PCR and WGS. These methods are well-described and established in the literature.

Progress 07/01/23 to 06/30/24

Outputs
Target Audience:The target audience dueing the first year of tge project included multiple stakeholders, including: 1- The poutry industry: We communicated our findings with two major poutry meat producers. This occurred during Zoom meetings and several conferences where thework was presented in poster format (The Center for Food Safety Annual Meeting, International Association for Food Protection (IAFP)). 2- The U.S. National Poultry Research Center in Athens: Our findings were communicated via Zoom meetings and also through active collboration. 3- The scientific community: Our findings were communicated in several local, national, and internatiomal meeting in the form of abstracts and posters. Changes/Problems:The only major challenge was the need for continous acccess to the poultry processing plant. Despite the full support of our industry partners, in the recent year there was a need for heightened biosecurity. Therefore, we had to collect the samples outside the plants rather than inside during operations. However, our industry partners are keen on providing continued access to test the interventions in the near future. What opportunities for training and professional development has the project provided?The project was used to train a PhD student, who performed the assigned experiments as part of a PhD thesis. The findings were presented by the PhD students in several local, national, and international meetings, allowing the student to hone presentation skills and interact with peers and seasoned scientists. How have the results been disseminated to communities of interest?The results were dissemnated via: 1- Zoom meetings with collboratores and stakeholders in the poutry industry. 2- The data were publised and presented in several scientific meetings. 3- A manuscript (that addresses objective 1) is being prepared for publication What do you plan to do during the next reporting period to accomplish the goals?We plan to continue to achieve our proposed objectives, with focus on objective 3: 1- We are testing different antimicrobials on artificiallycontaminated chicken meat to check their ability to significantly reduce process tolerant Campylobacter. This includes novel (niot tested previoulsy) phytochemicals.

Impacts
What was accomplished under these goals? We have achieved significant progress on this project. Objective 1: We have processed 122 chicken samples to isolate Campylobacter: We isolated 51 Campylobacter corresponding to 50 samples, which showed that the prevalence was at ~ 41.8%. The majority of the isolates (n = 42) were identified as C. jejuni (80.4%), while only 7 isolates were C. coli (13.7%). All 51 isolates were sequenced using Whole genome sequencing. Sequences were deposited in GenBank. Sequence types were determined for all the isolates. ST353 and ST48 were the most prevalent in C. jejuni, while ST7818 was most prevalent in C. coli. The antimicrobial resistance profiles were determined for all strains. We found that 65% of the isolates were resistant to ampicillin, 14% to ciprofloxacin, 22% to nalidixic acid, and 20% to tetracycline. We identified different antimicrobial genes and mutations in the isolates, including blaOXA alleles (encodes resistance to β-Lactams), aph(3') and sat-4 (encodes resistance to aminoglycosides), tet(O)( (encodes resistance to β-Lactams), and cmeR (multidrug efflux pump). Mutations in the gyrA that are associated with resistance to quinolones were also identified. We identified several virulence associated genes in the isolates, including cdtABC which was detected in 100% of the C. jejuni. Objective 2: We have found that relatively high concentrations of PAA and chlorine wererequired to effectively inhibit the Campylobacter strains (dubbed process tolerant strains). Each strain was tested individually against PAA and cholrine in lbuffer as well as on chicken meat. The strains that survived the treatments were designated as process tolerant. Different starins exhibited varying tolerance; some having notable high tolerance to these chemicals. Other tested organic acids were either similar or less effective when compared to PAA and chlorine.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: N. Nasser, T. Xu, D. Mann, X. Deng, I.I. Kassem. Prevalence and characterization of sequence types, antimicrobial susceptibility and virulence determinants of Campylobacter spp. isolated from retail poultry meat in Georgia, USA. Center for Food Safety (CFS) Annual Meeting, Atlanta, GA, 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: N. Nasser, I.I. Kassem. Evaluation of different organic acids for controlling multiple foodborne bacterial pathogens. Center for Food Safety (CFS) Annual Meeting, Atlanta, GA, 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: N. Nasser, I.I. Kassem. Comparative analysis of the impact of different organic acids against major foodborne bacterial pathogens. Association of Official Agricultural Chemists (AOAC) Southern Section, Atlanta, GA, 2023
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: N. Nasser, I.I. Kassem. In-depth assessment of Campylobacter spp. in retail chicken meat in Georgia, USA. Association of Official Agricultural Chemists (AOAC) Southern Section, Atlanta, GA, 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: N. Nasser, M. Barr, I.I. Kassem. Evaluation of different organic acids for controlling multiple foodborne bacterial pathogens. International Association for Food Protection (IAFP) Annual Meeting, Toronto, Canada, 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: N. Nasser, I.I. Kassem. Prevalence and comprehensive characterization of Campylobacter species isolated from poultry meat in retail stores in Georgia, USA. International Association for Food Protection (IAFP) Annual Meeting, Long Beach, CA, 2024.