Source: IOWA STATE UNIVERSITY submitted to
CAMRADES CONNECTING ANTIMICROBIAL RESISTANCE, AGRICULTURAL DECISIONS, AND ENVIRONMENTAL SYSTEMS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1028183
Grant No.
2022-68015-36717
Cumulative Award Amt.
$1,000,000.00
Proposal No.
2021-08949
Multistate No.
(N/A)
Project Start Date
Jan 15, 2022
Project End Date
Jan 14, 2027
Grant Year
2022
Program Code
[A1366]- Mitigating Antimicrobial Resistance Across the Food Chain
Project Director
Soupir, M. L.
Recipient Organization
IOWA STATE UNIVERSITY
2229 Lincoln Way
AMES,IA 50011
Performing Department
Ag & Biosystems Engineering
Non Technical Summary
Our project will inform the effectiveness of mitigation strategies in reducing risk to human health in agro-ecosystems by developing an adaptable framework, CAMRADES, Connecting Antimicrobial Resistance, Agricultural Decisions, And Environmental Systems. We will integrate predictive models of AMR transport and associated risk to human health, and improve stakeholder understanding of AMR, potential risks and mitigation strategies.Objective 1. Characterize and predict transport of AMR targets via surface and subsurface pathways in agro-ecosystems.Objective 2. Integrate results into a novel adaptable framework (CAMRADES) to assess risk associated with transport of AMR through agricultural environments to humans as a result of agricultural practices.Objective 3. Improve knowledge of AMR-related risks and inspire adoption of practices among food producers to combat AMR-related health and food safety risks associated with agro-ecosystems.Our framework integrates detection of antibiotics, AMR genes and indicator organisms and exposure pathways in the environment with hydrologic and water quality monitoring. A key output of this effort will be a predictive model of antibiotic and AMR sources, fate, and transport, which will be integrated with a novel risk assessment. Finally, to maximize realized AMR risk reduction, combinations of mitigation strategies will be informed by potential efficacy scenarios encompassing barn, manure, and land management strategies and disseminated using the recognized and trusted iAMResponsibleTM Project platform as well as Livestock Extension Programs. Our project will cumulate with an AMR Summit and AMR reduction strategy, a science and technology-based framework to reduce AMR in shared water resources. We will work closely with stakeholders to provide optimal flexibility to meet producer needs.
Animal Health Component
50%
Research Effort Categories
Basic
50%
Applied
50%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1120320202090%
1330210107010%
Goals / Objectives
Our goal is to assess the effectiveness of various mitigation strategies for reducing risk to human health in agro-ecosystems by developing an adaptable framework, Connecting AntiMicrobial Resistance Agricultural Decisions and Environmental Systems, CAMRADES. We will integrate predictive models of AMR transport and associated risk to human health, and improve stakeholder understanding of AMR, potential risks and mitigation strategies, along with motivating adoption of research-based practices to protect human health. Supporting ObjectivesCharacterize and predict transport of AMR targets via surface and subsurface pathways in agro-ecosystems.Integrate results into a novel adaptable framework (CAMRADES) to assess risk associated with transport of AMR through agricultural environments to humans as a result of agricultural practices.Improve knowledge of AMR-related risks and inspire adoption of practices among food producers to combat AMR-related health and food safety risks associated with agro-ecosystems.
Project Methods
3.0 Approach The foundation of the proposed work is the development of CAMRADES, which is built upon the synthesis of both gene, antibiotic, and bacterial datasets from monitoring efforts, the integration of AMR datasets into SWAT, and connecting AMR datasets to human health risks. In its implementation, CAMRADES will allow for the integration of both feasibility and mitigation efficacy and the evaluation of the impacts of different farm management decisions on the introduction and mitigation of AMR distribution. Finally, we propose to develop outreach tools to engage diverse producers, policy makers, and educators.Research activities in Obj. 1 will address our research question: How do agro-ecosystem practices influence human AMR risk via environmental pathways? To expand, the subsurface pathway reflects the potential of AMR amplification in the soil and movement into tile or groundwater while the surface pathway is runoff of AMR, which indicates the influence of certain land management decisions (such as manure application). Distinguishing between these pathways is important to understand the effectiveness of interventions, especially those implemented on the landscape. To accomplish this effort, we will build upon existing watershed monitoring and perform additional assessment of two watershed for a systematic study of AMR transport in agroecosystems.Watershed Monitoring Datasets. First, we will compile and expand on extensive monitoring of surface and drainage from two agriculturally dominant watersheds across different land regions in IA and NE for which historical antibiotic and AMR datasets are already available. 1) The Shell Creek watershed drains approximately 120,000 km2 in east-central NE and dominated by agricultural landuse: 46% of the land is in irrigated row crops and the watershed also contains over 1 million head of cattle, swine, and poultry 16 and 2) The BHL watershed is located along the western edge of the Des Moines Lobe landform of Central Iowa, with a drainage area of 5,324 ha and more than 80% of the watershed used for agricultural production including cattle and swine 13, 49, 50. BHL has a diverse range of management practice implementation. In both cases we are building on long-term monitoring projects which have included antibiotic and DNA collection and analysis, providing robust datasets for modification of the predictive model.Here, we propose to expand this effort to include ARG surveillance of a broad diversity of genes previously observed to be associated with manure inputs, allowing for lower limits of detection of AMR. Further, because we have preserved DNA from past monitoring efforts, we will re-run samples using the expanded high throughput qPCR methods described below. This effort parallels a long history of research investigating different management practices in agroecosystems to beneficially manage crop production and human health, with much of the focus being on nutrients, pesticides, and fecal indicator bacteria. Assessment of similar strategies on AMR mitigation is a rapidly developing field and sufficient data now exists at multiple intervention points to assess impacts and risk to human health.Data analysis: Statistical comparisons of resistance gene abundance and target gene analysis will be performed. Additionally, co-occurrence analysis to identify networks of genes that share similar patterns of abundance changes in different conditions (e.g., over time or with and without ARG history) will be used to determine networks of genes that may be correlated 71. Similar methods will be used to assess relationships between ARGs, indicator organisms, and antibiotic concentrations. All data will be tested for normality and transformed if necessary to meet assumptions of normality and equality of variances. Non-detects will be taken as ½ of the limit of detection for antibiotic data and qPCR assays. Correlation analysis will be used to evaluate relationships between AMR and environmental datasets.SWAT integration to predict AMR fate and transport (SWAT-AMR). The SWAT model includes a framework for predicting fecal indicator bacteria transport in agroecosystems, but it has not been developed to simulate AMR. Here we propose to build a new module in SWAT using publicly available management datasets combined with the proposed data collection on ARGs and ARBs in the BHL and Shell Creek watersheds. We will modify the existing SWAT FIB component to predict AMR using ARG and ARB water quality data. The outcome is SWAT-AMR, a watershed-scale model to predict movement of ARG and ARB through environmental pathways in agroecosystems.Risk Assessment Integration Our first step in Obj. 2 is to connect our biophysical model (SWAT-AMR) output (e.g. ARB, ARG) to a human risk assessment. In line with the proposal scope, the following aspects that may affect the risks attributable to antibiotic resistant genes derived from various environmental compartments will be evaluated, including (1) the likelihood of a gene being transferred to human bacteria, (2) the probability of disseminating the gene through the environmental exposure pathways, and (3) severity of the public health problem. The first two aspects determine the probability of exposure, while the last one relates to the severity of consequence. In turn, a number of risk descriptors effecting the above-mentioned aspects will be investigated.Scenario-based AMR risk/ reduction metrics. Next, we will integrate expected AMR reduction (as generated by review of existing practices and informed by our ongoing work) to develop realistic estimates of AMR reduction/transport under current and potential management scenarios. The estimated AMR reduction for a range of practices will be characterized by realm, Barn Management, Manure management, and Land Management. From this we will estimate the ratio of the expected percent AMR Reduction (%R) which will be generated for hundreds of potential scenarios using our CAMRADES model, converging environmental datasets with risk assessment to inform expected impact of management decisions on human health risk.Improve knowledge of AMR-related risks among food producers.In year 5, we will convene an AMR Summit that will bring key stakeholders together to evaluate project results and initiate the development of an AMR action plan. Summit participants will include the leadership of the major stakeholders from the agribusiness community, including producer groups (e.g., Pork Producers, Cattlemen, Farm Bureau), veterinary service providers, manure management technical service providers, and livestock integrators (e.g., JBS, Smithfield Foods). The overall goal of the AMR Summit will be to gain consensus on future actions to address AMR. We will discuss the potential development and implementation of an "AMR Reduction Strategy". These efforts will cumulate in a strategy document showing alternative means of achieving a set reduction in the quantity of AMR transported to waters (e.g., 50%).?

Progress 01/15/23 to 01/14/24

Outputs
Target Audience:Environmental Scientists, Engineers, Landowners who apply manure and install conservation practices, watershed managers, farmers (livestock and crop), state agents Changes/Problems:Detection of antibiotics has been infrequent and thus the team is considering adding passive samplers to select sites in the BHL and Shell Creek Watersheds. Samplers would be deployed for 2 week time periods and then extracted and sent to UB for analysis. What opportunities for training and professional development has the project provided?We currently have four graduate students involved with this project, 1 at Iowa State University, 2 at UNL, and 1 University of Buffalo. The students have been trained on field and laboratory methods related to data collection and analysis, leading or assisting with field data collection and laboratory analysis, and modeling. All graduate students attend the project meetings with PIs. How have the results been disseminated to communities of interest?Primary dissemination for the iAMResponsible team has been via social media, and other online publications, in course materials, and at professional, educational, and community events. A student presented the calibration results of SWAT+ model when compared to the lumped version of SWAT at the ASABE meeting in summer 2023. We are currently working to develop additional materials for dissemination to farmers in 2024. What do you plan to do during the next reporting period to accomplish the goals?The teams as ISU, UNL, and UB will continue data collection, refining laboratory methods, and testing and modifying the SWAT+ model for prediction of AMR in watersheds. In 2024 we will be potentially deploying passive samplers to identify low levels of antibiotics present in the watershed. Target genes will be finalized and samples analyzed for resistant genes using qPCR. The SWAT model of the BHL and Shell Creek watersheds will continue to be tested and calibrated. For the Risk Assessment, multi-decision-criteria approach will be applied for determining the weight of each criterion. Once the abundance of genes determined in the surveillance part, the risk index of each sample will be determined incorporating the severity and the abundance of corresponding ARG. Looking further ahead, we will continue to include the severity to provide a risk assessment sheet including relevant questions that are likely to increase or decrease the risk for user applications. During the next reporting period the iAMResponsible team will continue to expand our content library to incorporate and make accessible research findings from a wider selection of contributors and expand our content creation to new media types and delivery methods. We will present Livestock and Poultry Environmental Learning Community (LPELC) webinars covering treatment of manure, fate and transport of AMR (in the environment), and risk assessment advancements. Three students will be presenting research talks or posters at upcoming professional conferences (ASEE, EDAR, and IAFP).

Impacts
What was accomplished under these goals? Supporting Objective 1. Characterize and predict transport of AMR targets via surface and subsurface pathways in agro-ecosystems. Characterization of antimicrobial targets at the Black Hawk Lake and Shell Creek Watersheds continued this year. Monitoring in the Black Hawk Lake (BHL) Watershed began in March and ended in November 2023.Sample collection and processing went as scheduled, i.e., every two weeks, with added sampling for storm events. In Iowa, this year was classified as dry when compared to the 30-year average and continues a drought period that since the initiation of the project. A total of 65 samples were collected from BHL sites, of which 61 were biweekly baseflow, and 4 were storm samples. All samples were processed for antibiotics, bacteria, and DNA. The Standard Operating Procedure (SOP) for antibiotic processing was updated and collaborators are continuing to work to improve antibiotic recovery. Following the updated SOPs, samples for antibiotics were processed through solid-phase extraction and shipped to the University at Buffalo (UB) for further processing. Total and resistant bacteria from water samples were grown on the selective agar, and DNA on filters was preserved in DNA bead tubes, stored at -80°C. Collaborators held multiple online meetings this year to keep track of the progress. Results of bacteria and antibiotics analysis for 2022 and 2023 were presented and discussed during the end-of-the-year meeting and uploaded to the common project database. SWAT+ models of BHL and Shell Creek watersheds was developed in the past year. The BHL site model was successfully calibrated for simulating water quantity and efforts are ongoing to calibrate water quality. A conference presentation of this model was made at the ASABE Annual International Meeting in July 2023. The model is currently undergoing nutrient and bacteria calibration, after which the team will move toward calibrating the model for antibiotics. Improve knowledge of AMR-related risks and inspire adoption of practices among food producers to combat AMR-related health and food safety risks associated with agro-ecosystems. To create a risk assessment model in line with Objective 3, we began by reviewing existing literature and identified four distinct criteria for assessing the severity of each Antibiotic Resistance Gene (ARG). These criteria include gene mobility, human accessibility (the gene's presence in various environments), pathogenicity (its presence in human pathogens), and resistance to clinically important antibiotics. We collaborated with Dr. Nielsen from the University of Copenhagen to obtain mobility data for genes, which had previously been extracted in a separate study. Ongoing work is on evaluation of the presence of genes in different environments using metagenomics information from accessible datasets.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Ahmed, S., S. Azzam, A. Syed, M.L. Soupir. 2023. Comparison between SWAT and SWAT+ for simulating streamflow in a small, heavily tile -drained agricultural watershed. ASABE Annual International Meeting in Omaha, NE. 7/9  7/12/2023.


Progress 01/15/22 to 01/14/23

Outputs
Target Audience:Environmental Scientists, Engineers, Landowners who apply manure and install conservation practices, watershed managers, farmers (livestock and crop), state agents Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We currently have fourteen studentsstudents involved with this project, 3 at Iowa State University, 10 at UNL, and 1University of Buffalo. The students have been trained on field and laboratory methods related to data collection and analysis, leading or assisting with field data collection and laboratory analysis, and modeling. All graduate students attend the project meetings with PIs.During the reporting period 1 PhD student, 2 masters students, and 4 undergraduate students developed or refined STEM communication skills by contributing to the development and distribution of educational materials for the iAMResponsible networks. The continued expansion of media development by the iAMResponsible team has also required several team members to develop new expertise in video production, audio production, and graphic design, utilizing programs such as Canva, and the Adobe Creative Suite, among others. How have the results been disseminated to communities of interest?Primary dissemination for the iAMResponsible team has been via social media, and other online publications, in course materials, and at professional, educational, and community events. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period the iAMResponsible team will continue to expand our content library to incorporate and make accessible research findings from a wider selection of contributors and expand our content creation to new media types and delivery methods. The teams as ISU, UNL, and UB will continue data collection, refining laboratory methods, and testing and modifying the SWAT+ model for prediction of AMR in watersheds.

Impacts
What was accomplished under these goals? Characterize and predict transport of AMR targets via surface and subsurface pathways in agro-ecosystems. Characterization of antimicrobial targets at the Black Hawk Lake and Shell Creek Watershed began this year. Monitoring in the Black Hawk Lake (BHL) Watershed began in March and ended in November 2022. Samples were collected every two weeks and during storm events. A total of 98 samples were collected at sites within the BHL monitoring network. Of the samples collected, 65 were at base flow and 21 at elevated storm flows. Monitoring in the Shell Creek Watershed began in October and included one sampling event at base flow. Each sample collected from this year was processed for antibiotics, bacteria, and DNA within 24 hours. The collected water samples underwent solid phase extraction (SPE) at Iowa State University (ISU) before SPE cartridges were then individually wrapped in aluminum foil and shipped on ice to University at Buffalo (UB) for further analysis. Bacteria from the water samples were grown on selective agar to measure total and resistant fecal indicator bacteria. Finally, the water was filtered for DNA and the filters were stored at -80°C until subsequent DNA extraction for antibiotic resistance gene quantification. To improve synergistic performance across institutions, collaborators from the University of Nebraska-Lincoln (UNL) toured the BHL watershed in August where the Iowa State University hosts shared their field and laboratory protocols. Additionally, the ISU collaborators toured the Shell Creek Watershed in October.Historic data of the BHL watershed was revisited, conditioned and a common data space was setup. Observed data collected from this year was acquired, formatted and made available in the project database. Database sharing modalities between ISU and UNL are being discussed and a platform will be selected for rapid data sharing. Integrate results into a novel adaptable framework (CAMRADES) to assess risk associated with transport of AMR through agricultural environments to humans as a result of agricultural practices. ISU also initiated modeling of the BHL watershed on SWAT+. Datasets like the DEM, climatic variables, soil and landcover to be used in modeling (other than the observed water quality and quantity) were explored for simulating BHL watershed on SWAT+. The model is currently under the calibration phase at a sub watershed level. Improve knowledge of AMR-related risks and inspire adoption of practices among food producers to combat AMR-related health and food safety risks associated with agro-ecosystems. The iAMResponsible team works to translate and transfer new research findings for delivery to the target audiences. Over 50 new pieces of outreach content were created during the reporting period for distribution on social media and added to the existing database of media and research related to AMR curated by the iAMResponsible team and available to the public on the Livestock and Poultry Environmental Learning Community (LPELC) website. Four on-line social media outlets for dissemination of AMR related materials are managed with regular (monthly, weekly, or daily) outputs. In partnership with LPELC the project team delivered the fourth in an ongoing series of webinars on AMR impacts on agriculture aimed at agricultural producers and their advisors. In early 2022, the iAMResponsible team launched a podcast series (Tales of the Resistance), to discuss research findings on AMR in food production systems with general (non-expert audiences). The iAMResponsible team is also leading a multi-university online course "AMR from a One Health Perspective" which explores the many faceted challenges of AMR with emphasis on developing scientific communication skills in young STEM professionals. During the Spring of 2022, twenty students at six universities (Nebraska, Maryland, North Carolina State, Washington State, Oklahoma State, and Minnesota) participated in the course.

Publications