Source: TENNESSEE STATE UNIVERSITY submitted to NRP
A MULTI-FACETED APPROACH TO MITIGATE ANTIBIOTIC RESISTANCE IN HISTORICALLY UNDERSERVED POULTRY FARMS BUDGET AND BUDGET NARRATIVE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1032403
Grant No.
2024-69015-42442
Cumulative Award Amt.
$486,310.00
Proposal No.
2023-10486
Multistate No.
(N/A)
Project Start Date
Jul 1, 2024
Project End Date
Apr 25, 2025
Grant Year
2024
Program Code
[A1366]- Mitigating Antimicrobial Resistance Across the Food Chain
Recipient Organization
TENNESSEE STATE UNIVERSITY
3500 JOHN A. MERRITT BLVD
NASHVILLE,TN 37209
Performing Department
(N/A)
Non Technical Summary
The extensive use of antibiotics in livestock farming poses a significant public health risk with widespread consequences. Animal agriculture generates considerable waste, fostering the rapid growth and spread of disease-causing pathogens in the environment. To combat this, farmers often resort to using medically important antibiotics to control the spread of diseases among their livestock. However, this practice exposes both harmless and harmful bacteria to antibiotics, leading to the emergence and dissemination of antimicrobial resistance (AMR). Consequently, the farming environment becomes a breeding ground for antibiotic-resistant bacteria, which can then infect humans either through direct contact or the food chain. Recognizing that food animals not only transmit but also contribute to the proliferation of antibiotic-resistant bacteria, many countries have implemented stricter regulations on antibiotic use in livestock farming. These regulations typically restrict antibiotic usage and mandate veterinary supervision. The US government, for instance, has imposed limitations on antibiotic usage in animal farming to combat the overuse and misuse of these drugs, which can spawn antibiotic-resistant strains. However, it's crucial to acknowledge the challenges faced by historically underserved farmers in complying with these regulations. Such farmers often encounter unique obstacles such as disease management, water contamination, limited access to veterinary services, and inadequate resources and education on best practices, all of which can impact the prevalence of poultry diseases and the spread of AMR.The overarching goalof this projectis to establish an educational and training program on antimicrobial stewardship, specifically tailored to empower historically underserved poultry producers. By imparting knowledge and fostering practices such as good farming techniques, biosecurity measures, and the establishment of strong Veterinarian-Client-Patient Relationships, this willreduce the incidence and spread of poultry diseases while minimizing antimicrobial usage on poultry farms. Additionally, the project seeks to introduce innovative solutions such as UV light combined with peracetic acid to combat microbial contamination and biofilm formation in key areas such as water systems. UV-C irradiation will also be utilized to control microbial contamination on egg surfaces. The project also aims to enhance the capabilities of Extension educators and producers in poultry diseases and antimicrobial resistance through workshops.Well-trained Extension personnel will serve as conduits for transferring knowledge to producers and effectively engaging with their clientele. By simplifying complex topics related to poultry diseases and antimicrobial resistance, they will empower historically underserved poultry producers with valuable insights and information.
Animal Health Component
70%
Research Effort Categories
Basic
20%
Applied
70%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
30732991100100%
Knowledge Area
307 - Animal Management Systems;

Subject Of Investigation
3299 - Poultry, general/other;

Field Of Science
1100 - Bacteriology;
Goals / Objectives
The emergence of antimicrobial-resistant pathogens in animal production systems is a significant challenge to public health. The U.S. government has placed guidelines to reduce antibiotic use in animal farming. However, it's essential to acknowledge the challenges that historically underserved producers may face in meeting these requirements. There are no alternatives to antibiotics (ATA) to curtail the spread of diseases on farms, hence historically underserved poultry producers have been left in unfamiliar territory. Historically underserved producers often face unique challenges stemming from limited resources, risk farming practices, unable to access information on disease management, and minimum veterinary technical support, hence the need for Veterinarian Client-Patient Relationship (VCPR). Lack of access to information and education on poultry husbandry continues to be evident in Historically underserved poultry. Education and awareness are the primary tools to change inappropriate antimicrobial use, confront livestock diseases and AMR in production systems. Knowledge gap can lead to inefficient resource utilization, disease outbreaks, and suboptimal production yields. Equipping these farmers with the knowledge and skills they need is not only essential for their productivity and profitability, but also plays a crucial role in ensuring the welfare of the birds and maintaining a safe food supply.Therefore, the overall goal of this project is to establish an antimicrobial stewardship educational and training program for historically underserved poultry producers, enabling them to implement effective antimicrobial resistance (AMR) management practices. The objectives of this project are:1. Determine and antimicrobial-resistant bacteria (ARB) and antimicrobial-resistant bacteria genes (ARGs) in historically underserved poultry farms. This offers microbial diversity, ARB, ARGs profiles that provide contextual and background data relevant to mitigation of antimicrobial resistance. In addition, Resistance Maps, an interactive collection of charts and maps will be generated to it easier for producers and the public to understand antimicrobial resistance in animal production.2. Introduce UV light in combination with peracetic acid at critical on-farm control points to control microbial contamination and biofilm formation. This will control microbial contamination on the farm and significantly lower the risk of contamination. This contributes to safer and healthier food for consumer.3. Establish valid Veterinarian-Client-Patient Relationships (VCPRs) in historically underserved poultry farms. VCPRs are essential to ensure proper diagnosis and disease treatment; explore producers' beliefs, antimicrobial practices, and occurrence of resistant microorganisms in in historically underserved poultry farms.4. Build a capacity of Extension educators and producers knowledgeable in poultry diseases and antimicrobial resistance. Well-trained Extension Specialists will communicate complex topics related to poultry diseases and antimicrobial resistance in a clear and understandable manner to historically underserved poultry producers. Through the project, historically underserved poultry farms will be assisted and supported as they navigate the challenges associated with diseases and antimicrobial resistance (AMR), responsible antibiotic use, and best management practices BMPs. Farmers will be empowered to make informed and responsible decisions, which will have a ripple effect in the agricultural community. As a result, producers will not only contribute to the U.S. government effort to combat AMR, but also maximize their profits, ensuring long-term economic sustainability within their production systems.
Project Methods
Approach to meet objective 1: Environmental samples will be collected from 120 historically underserved poultry farms (East TN=40, Middle TN = 40; West TN = 40). Poultry farms (n=10) from five counties from each region will be randomly selected for sample collection. Samples (soil = 840; manure =840, water n=840; rectal n= 840) will be collected quarterly (seven times). Basic information including type of birds, time and place of collection will be recorded to permit more inclusive analysis of laboratory data, assist clarify potential biases for different sample types, and help ascertain critical control points for mitigating resistance. DNA will be extracted for metagenomics studies using Bio PowerFecal®, PowerWater, and PowerSoil® DNA isolation Kits. Manure and water samples will be aseptically sequenced by Illumina HiSeq and analyzed by using EzBiome Lab Bioinformatics Package. Briefly, gDNA will be enzymatically sheared, fragment ends will be repaired, 3' adenylated, and ligated to adapters according to the manufacturer's instructions. The adapter-ligated libraries will be PCR-amplified and cleaned up from the reaction mix with Mag-Bind RxnPure Plus magnetic beads. The libraries will then be quantified using the D1000 ScreenTape on an Agilent 2200 TapeStation instrument. The qualified libraries will be normalized and pooled for multiplexed sequencing on an Illumina HiseqX sequencer (Illumina, San Diego, CA, USA) using the pair-end 150bp run format targeting 40-50 M read depth per sample. Furthermore, will incorporate spike-in controls representing three microorganisms with known abundances into all samples to serve as internal references, allowing us to accurately quantify the abundance of taxa and AMR genes across samples. Shotgun DNA sequences will be taxonomically and functionally profiled and compared by EzBiome bioinformatics algorithm and curated databases. In addition to metagenomic sequencing, there will be complement of analysis with targeted qPCR analysis on a subset of samples for the most abundant and prevalent AMR genes.Approach to meet objective 2: Implementation of UV treatment system and peracetic acid in small poultry farm: Commercially available UV-C water disinfection devices (Sanitron S2400C NSF Certified (40 GPM) and UV-25T80; SARIN Energy Solutions (130GPM)) alongside peracetic acid (PAA) treatment (80 ppm via Venturi Injection System) will be integrated to in-line water distribution line to implement the proposed intervention for enhancing the safety of water used on the farm. The treatment water will pass through the UV-C device equipped with venturi system for peracetic acid treatment (optimized for delivery of 80±5ppm sanitizer concentration) and exit the UV device with potentially reduced contamination levels and ready for use by the birds. Farms will be visited to collect water samples before and after treatment using UV light and peracetic acid and transported to Food Microbiology Lab at Tennessee State University under refrigerated conditions.Biofilm Studies in the laboratory:For the purpose of studying biofilms in water lines in farms, a CDC Biofilm Reactor will be employed. The reactor features a 1-liter glass container with a maximum liquid capacity of 500 ml. It is equipped with eight detachable rods, each holding three coupons with a diameter of 1.27 cm, which will act as the substrate for biofilm growth.Developing the decision-making tool:A mathematical model (decision-making tool) will be developed to educate and train producers and extension agents on the application of the technology tested. The tool can be accessed athttps://trevorhefley.shinyapps.io/uv-1/.Introduce to producers the UV-C irradiation on egg surfaces:Since complex microbial and in biofilms develop resistance to current physical and chemical disinfection methods, a novel UV-C LED systemwill be used for egg exposuresto disinfect surface contamination on the egg shells.Approach to meet objective 3:Establish valid Veterinarian-Client-Patient Relationships (VCPRs). This objective will be achieved through three steps:Step I: Collect information on poultry diseases, management, actual antimicrobial use, biosecurity, disposal practices, and evidence of veterinarian visits or consultation in historically underserved poultry farms.Step II:Veterinarian-Client-Patient Relationship (VCPR):12 VCPR workshops and 3 webinars will be delivered as follows: During Year one: 6 workshops (East TN =2; Middle TN=2; West TN = 2) and one (n=1) webinar will be conducted; Year 2:6 workshops (East TN =2 Middle TN=2; West TN = 2) and one (n=1) webinar will also be conducted. Year 3, one workshop for all producers will be held at the Tennessee State University Research and Education Center. In addition, one (n=1) webinar will also be conducted in year 3. Workshops on VCPR will be conducted to educate producers the importance of VCPR and also know their beliefs and diseases management practices.Approach to meet objective 4:Build a capacity of Extension educators and producers knowledgeable in poultry diseases and antimicrobial resistance.Year 1 of the project: Three webinars will be conducted: Webinar #1: Focus on Pathogenic Bacteria and Antimicrobial Resistance on Farms, designed for participation by farmers, extension agents, and veterinarians from East, Middle, and West regions of Tennessee; 2. Webinar #2: Centered around Poultry Diseases, tailored for farmers, veterinarians, and extension agents; 3. Webinar #3: Highlighting Best Management Practices, with participation open to farmers, veterinarians, and extension agents.Year 2 of the project. Three Workshops: Focused on Pathogenic Bacteria, Antimicrobial Resistance, and Challenges Faced by Producers, will be held separately in the East, Middle, and West regions of Tennessee.There will also be a Trainer-the-Trainer Workshop: A one-day event providing a solid foundation in Best Management Practices (BMPs), risk assessment for minimizing poultry diseases, and addressing challenges related to antimicrobial resistance.Year 3 of the project: Developing an AMR Management Plan for Low-Resource Poultry Farms. Participants will include poultry producers, extension educators, veterinarians, consumers, students, faculty in agriculture, pharmaceutical representatives, policy makers, and government officials from the Department of Agriculture. Following the workshop, a structured questionnaire will be administered to measure knowledge gained, as well as changes in behavior and attitude towards AMR management.Students' involvement in the project:Students will be engaged in research, educational antimicrobial stewardship workshops which will provide an unparalleled experiential learning opportunity. Graduate students will receive extensive training in molecular and microbiological techniques. In this project, graduate students will also conduct their thesis and research papers and attend and present research and extension findings at professional conferences.

Progress 07/01/24 to 04/25/25

Outputs
Target Audience:The primary target audience for this project includes poultry farmers who face unique challenges related to antimicrobial resistance (AMR). These farmers often operate in high-risk farming practices, and dont have information on biosecurity infrastructure. The project also targets agricultural extension agents, veterinarians, and other stakeholders involved in poultry health management and sustainable agriculture. By focusing on this audience, the project aims to deliver sustainable, applied, and informed solutions tailored to their specific needs, ultimately enhancing biosecurity practices, improving AMR awareness, and promoting the long-term viability of poultry farms. Changes/Problems:Major Changes/Problems in Approach and Reasons for Changes The project was terminated and did not continue to completion as originally planned. As a result, all planned activities were discontinued, and further data collection, outreach, and analysis were halted. The project was officially terminated due to circumstances beyond the control of the project team. This resulted in the cessation of research and extension activities associated with the original scope of work. No further modifications to the approach were implemented following the termination, and no additional results or outcomes were generated after that point. What opportunities for training and professional development has the project provided?Two undergraduate students had justactively started in data collection, laboratory procedures, and outreach activities, contributing to workforce development in agricultural sciences. The undegraduate students had just started working in the following studies (1). A comparative study assessing pathogen prevalence in backyard/roaming versus caged poultry production systems. (2) An evaluation of Staphylococcus aureus and Salmonella occurrence across different poultry environments to provide insight into microbial risks associated with production systems. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Objective 1: Determine and antimicrobial-resistant bacteria (ARB) and antimicrobial-resistant bacteria genes (ARGs) in historically underserved poultry farms;To achieve this objective, the following activities were conducted: 1. Environmental samples were systematically collected from poultry farms across Tennessee, distributed as follows: East TN (n=8), Middle TN (n=16), and West TN (n=6). For each sample, metadata including bird species, sampling date, and geographic location were recorded to facilitate comprehensive laboratory analyses and spatial AMRmapping.Geographical Information Systems (GIS) technology was employed to coordinate sampling sites and to be used to generate spatial maps illustrating the distribution of antimicrobial resistance (AMR). This geospatial approach serves as an effective tool to communicate AMR risks and clustering to poultry producers, addressing the critical need for clear dissemination of AMR information. Sample processing using selective media: Listeria Selective Agar (LSA) for Listeria monocytogenes, Campylobacter Blood-Free Selective Medium for Campylobacter, XLD agar (Xylose Lysine Deoxycholate agar) for Salmonella spp , Eosin Methylene Blue (EMB) agar for Escherichia coli, and XLD agar for Shigella spp. For each collection event, triplicate samples of soil, manure, and water were obtained, totaling 90 samples per matrix to date. Samples were screened for the presence of Listeria monocytogenes, Campylobacter spp., Salmonella spp., Escherichia coli, and Shigella spp. using selective culture media. Presumptive isolates from manure, soil, and water were archived at -80°C for future downstream molecular and resistance profiling analyses. 2. A series of online workshops were held covering key topics related to poultry biosecurity and foodborne pathogens. Producers were introduced to innovative antimicrobial interventions, specifically the application of ultraviolet (UV) light combined with peracetic acid treatments, targeted at critical on-farm control points to mitigate microbial contamination, and inhibit biofilm formation. 3. A pre were partly developed and to be ditributed to farmers and Extesion agents. Preliminary findings using bacteria selective agars:Producers (N=35) were introduced to best management practices innpoultry production. Analysis of environmental samples revealedfoodborne pathogens across different matrices. In manure samples, Listeria monocytogenes was detected in 3.2% of samples, Campylobacter in 0.8%, Salmonella in 1.6%, Escherichia coli in 86.5%, and Shigella was not detected. Water samples showed higher prevalence rates with Listeria monocytogenes detected in 42.5%, Campylobacter in 53%, Salmonella in 9%, E. coli in 75%, and no Shigella detected. Soil samples demonstrated Listeria monocytogenes presence in 45% of samples, Campylobacter in 0.2%, Salmonella in 0.5%, E. coli in 64%, and Shigella was absent.

Publications