FY23 NP Antimicrobial Resistance RFA MAY FINALV2_508.pdf

OVERVIEW INFORMATION

U.S. Environmental Protection Agency Office of Science Advisor, Policy and Engagement Office of Research and Development

NATIONAL PRIORITIES: EVALUATION OF

ANTIMICROBIAL RESISTANCE IN WASTEWATER

AND SEWAGE SLUDGE TREATMENT AND ITS

IMPACT ON THE ENVIRONMENT

This is the initial announcement of this funding opportunity.

Funding Opportunity Number: EPA-G2023-ORD-F1

Assistance Listing Number: 66.511

Solicitation Opening Date: May 19, 2023 Solicitation Closing Date: August 16, 2023: 11:59:59 pm Eastern Time

Table of Contents

I. FUNDING OPPORTUNITY DESCRIPTION

A. Introduction

B. Background C. Authority and Regulations D. Specific Research Areas of Interest/Expected Outputs and Outcomes E. References F. Special Requirements G. Additional Provisions for Applicants Incorporated into the Solicitation

II. AWARD INFORMATION

III. ELIGIBILITY INFORMATION

A. Eligible Applicants B. Cost Sharing C. Other

IV. APPLICATION AND SUBMISSION INFORMATION

A. Grants.gov Submittal Requirements and Limited Exception Procedures B. Application Package Information C. Content and Form of Application Submission D. Submission Dates and Times E. Funding Restrictions

F. Submission Instructions and Other Submission Requirements

V. APPLICATION REVIEW INFORMATION

A. Peer Review B. Relevancy Review C. Past Performance History Review D. Human Subjects Research Statement (HSRS) Review E. Evaluation of the Scientific Data Management Plan F. Funding Decisions

VI. AWARD ADMINISTRATION INFORMATION

A. Award Notices B. Administrative and National Policy Requirements

VII. AGENCY CONTACTS

I. FUNDING OPPORTUNITY DESCRIPTION

For Updates and Additional Information see https://www.epa.gov/research-grants/research-funding-opportunities.

View research awarded under previous solicitations at https://www.epa.gov/research-grants/research-grant-areas.

A. Introduction

In recent years, concern has increased about the occurrence of antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARGs) that could make it more difficult to treat certain infections in animals and people. Wastewater treatment facilities are believed to be potential receptors and sources for ARB and ARGs and can act as a bridge to the environment. This RFA seeks research that will address knowledge gaps in the occurrence, fate and transport, and persistence of antimicrobial resistant organisms and genes found in municipal wastewater effluent and biosolids. The RFA also requests research on combined sewer overflows, septic systems, and small wastewater systems. Research should provide a better understanding of the impact of ARB and ARGs on receiving waters including information on the relative contribution of ARB and ARGs from wastewater as compared to other sources such as animal agriculture, animal husbandry, hospital sources, and direct industrial sources. Research should also support the development of frameworks and methodologies for quantifying risk related to AMR in treated wastewater discharge, water reuse, and biosolids (and biosolids products) land application and beneficial use. Research results should improve our understanding of the nature, extent, selection, and removal of ARB and ARGs found in municipal wastewater effluent and biosolids.

The Office of Research and Development’s (ORD) Consolidated Research/Training/Fellowships program supports research and development to: (1) determine the environmental effects of air quality, drinking water, water quality, hazardous waste, toxic substances, and pesticides; (2) identify, develop, and demonstrate effective pollution control techniques; (3) perform risk assessments to characterize the potential adverse health effects of human exposures to environmental hazards; and (4) facilitate training and program participant support in these areas.

https://www.epa.gov/research-grants/research-funding-opportunities https://www.epa.gov/research-grants/research-funding-opportunities https://www.epa.gov/research-grants/research-grant-areas https://www.epa.gov/research-grants/research-grant-areas

Awards made under this program further EPA’s priorities supporting robust science for air quality, safe and sustainable water resources, sustainable and healthy communities, chemical safety, and human health risk assessment. The national priorities competition under this program supports high-priority water quality and availability research.

EPA recognizes that it is important to engage all available minds to address the environmental challenges the Nation faces. At the same time, EPA seeks to expand the environmental conversation by including members of communities which may have not previously participated in such dialogues to participate in EPA programs. For this reason, EPA strongly encourages all eligible applicants identified in Section III, including minority serving institutions (MSIs), to apply under this opportunity.

For purposes of this solicitation, the following are considered MSIs:

1. Historically Black Colleges and Universities, as defined by the Higher Education Act (20 U.S.C. § 1061(2)). A list of these schools can be found at Historically Black Colleges and Universities;

2. Tribal Colleges and Universities (TCUs), as defined by the Higher Education Act (20 U.S.C. § 1059c(b)(3) and (d)(1)). A list of these schools can be found at American Indian Tribally Controlled Colleges and Universities;

3. Hispanic-Serving Institutions (HSIs), as defined by the Higher Education Act (20 U.S.C. § 1101a(a)(5)). A list of these schools can be found at Hispanic-Serving Institutions;

4. Asian American and Native American Pacific Islander-Serving Institutions; (AANAPISIs), as defined by the Higher Education Act (20 U.S.C. § 1059g(b)(2)). A list of these schools can be found at Asian American and Native American Pacific Islander-Serving Institutions; and

5. Predominately Black Institutions (PBIs), as defined by the Higher Education Act of 2008, 20 U.S.C. 1059e(b)(6). A list of these schools can be found at Predominately Black Institutions.

B. Background

The genetic adaptability of microbes along with the prevalence of antimicrobial medicines have created a global health problem due to ARB and ARGs that could make it harder to treat certain infections in animals and people. Given rapid evolution and selection, ARB and ARGs can move between the environment, humans, and animals, which makes predicting where and when resistance occurs difficult. Antimicrobial resistance (AMR) has been identified by the World Health Organization (WHO) as one of the greatest threats to human health. Furthermore, both WHO and the U.S. Centers for Disease Control (CDC) have identified antibiotic resistance as a high priority research area. To further illustrate, according to a 2019 CDC report, every year 2.8 million resistant infections in the United States cause 35,900 deaths. From an economic https://sites.ed.gov/whhbcu/one-hundred-and-five-historically-black-colleges-and-universities/ https://sites.ed.gov/whhbcu/one-hundred-and-five-historically-black-colleges-and-universities/ https://sites.ed.gov/whiaiane/tribes-tcus/tribal-colleges-and-universities/ https://sites.ed.gov/whiaiane/tribes-tcus/tribal-colleges-and-universities/ https://sites.ed.gov/hispanic-initiative/hispanic-serving-institutions-hsis/ https://www.google.com/maps/d/viewer?mid=1XVkOWKMDORm53pvU0L8EPsrJC94&msa=0&ie=UTF8&t=m&z=3&source=embed&ll=40.58644586187277%2C-148.28228249999984 https://www.google.com/maps/d/viewer?mid=1wlIi3j7gtlNq_w-0NKAb2bF2VmY&ie=UTF&msa=0&ll=37.35160769312532%2C-96.17229800000001&z=4 standpoint, a direct estimate for treating six, common, multidrug-resistant pathogens is $4.6 billion a year (CDC, 2019). Given the scale and complexity of the problem, a “One Health” approach which recognizes the interdependencies between people and animals, but also acknowledges the importance of the ecosystems and environment, has been put into practice by multiple health organizations internationally (CDC, 2022). This approach emphasizes strong collaboration between animal and human health sectors as well as collaboration between different disciplines at different scales (regional, national, international) to prevent and control emerging diseases. Within the one health approach, the natural and engineered water environments have increasingly been shown to play significant roles in AMR evolution and spread. A recent National Academy of Sciences report (NAS, 2022) emphasized the need for improved guidance for assessing antimicrobial resistance in discharges from wastewater treatment given their importance as a link between anthropogenic contamination and the environment.

Wastewater treatment facilities are believed to be one of the major potential receptors and sources for ARB and ARGs. These facilities typically discharge to aquatic environments and act as both a barrier as well as a potential source of pollutant discharge to the environment. They receive a mix of pathogens, resistant genes, and antimicrobial drug residues from multiple points in the collection system, including wastewater from industry, households, and hospitals, all of which contribute to a high density of pathogens that enter the plant. The selective pressure of residual antimicrobial drugs coupled with gene transfer between microorganisms at the plant can potentially lead to the proliferation of ARB and ARGs during wastewater treatment processes.

Operational conditions in treatment processes may promote the proliferation of ARB resulting in higher fractions in treated wastewaters relative to raw wastewater or in finished biosolids or biosolids products, which are ready to be applied to land.

Despite recent studies investigating treatment, operational conditions, and environmental occurrence of AMR (Gerrity et al., 2018, Wengroth et al., 2021), further information is needed to characterize the occurrence and significance of AMR found in treated and discharged municipal wastewater effluent, especially effluent used for reuse applications. Reported AMR selection and removal efficiencies in wastewater treatment systems have been highly variable, most likely due to the impact of the treatment mechanism, operational parameters and setting, and other still unknown factors. New research and controlled studies are needed to better understand the key technology (conventional and advanced); operational and site-specific parameters that affect AMR occurrence and growth, such as wastewater parameters, antimicrobial concentrations, operational conditions, dry or wet weather flow; and mechanisms responsible for attenuation such as photo-reactivation. Similarly, significant knowledge gaps exist on the potential occurrence, mechanism, and timing of genetic mobilization of ARGs between microbial communities in wastewater reclamation facilities and the human health implications that may be present in potable water reuse. The conditions that result in horizontal gene transfer of ARGs from bacteria in the environment to pathogenic bacteria found in clinical infections are also not well understood. Research in these areas should support development of effective mitigation measures.

Along with treated effluent, sewage sludge has the potential to concentrate significant amounts of ARB and ARGs from sewage. Sludge processing prior to land application or use as fertilizer includes multiple unit processes to reduce pathogens and vector attraction. Biosolids, the resulting byproduct from sludge, is, in many cases, applied to land where it can potentially spread ARB and ARGs into the environment. A better understanding of the impact of various in-plant biosolids treatment processes on the fate of AMR in biosolids is needed. Furthermore, research on potential new biosolids management mitigation practices may be needed to reduce or remove AMR within treatment processes. The transfer of ARGs found in biosolids applied to soils and harvested crops as well as from the use of biosolids products such as compost and heat-treated pellets has also not been well-investigated. In addition, more information is needed on the horizontal transfer of mobile ARGs (resistome) in biosolids to human pathogens and commensals and their potential role in the spread of AMR from the environment to human pathogens (Law et.al., 2021). Such research may inform the potential development of new management practices to protect public health.

Other sources of ARB and ARGs reaching the environment due to wastewater treatment processes include combined sewer overflows (CSOs) and non-point sources such as septic systems. Untreated sewer sediments and wastewater containing ARB and ARGs may be discharged to surface water during storm events (Eramo et al., 2017). Similarly, aging or inadequate wastewater collection systems along with septic systems are potential non-point sources of ARB and ARGs to the environment (Damashek et al., 2022). More information on non-point sources and CSOs is needed to determine the relative contributions of AMR from these sources.

As treated wastewater is typically discharged to aquatic environments, these environments are potential exposure routes to transmit resistant pathogens and genes to humans and animals via irrigation, recreational use, or drinking water exposure. Furthermore, wastewater biosolids, manure, surface water, and reclaimed wastewater are routinely applied to crops and agricultural land, exposing soils and plants to the antimicrobials, ARB, and ARGs. Soils can serve as reservoirs of AMR, facilitating the maintenance and spread of ARGs within their microbiome and among plant-associated bacteria (Christou et al., 2017; Tyrell et al., 2019). Vegetable crops can become contaminated through contact with the soil or directly via irrigation water, thereby serving as a potential route of human exposure (Hölzel et al., 2018). Recreational contact with contaminated surface waters may also result in AMR infections, as evidenced by epidemiological studies demonstrating elevated ARB colonization among exposed groups (Leonard et al. 2018; Søraas et al. 2013). Contaminated drinking water has likewise been associated with increased risk of ARB carriage (Coleman et al., 2012). Although drinking water treatment processes are generally effective in reducing ARB and ARGs, both have nevertheless been detected in treated drinking water (Sanganyado and Gwenzi 2019).

Given the multiple exposure routes for AMR, knowledge is needed to inform public health and environmental impact implications and to establish appropriate frameworks for quantifying risk related to AMR in treated wastewater discharge, water reuse, and land applied biosolids. In addition, it is not clear if, and to what extent, the level of AMR in the environment is due to the presence of AMR in the raw sewage influent (including those that receive untreated hospital waste), the proliferation of AMR during wastewater treatment, subsequent gene transfer in the environment, and other AMR sources that are completely unrelated to municipal wastewater such as animal agriculture or husbandry, direct discharges, and commercial or industrial sources.

As such, the significance of the discharge of wastewater and use of biosolids on the presence of AMR should be assessed and compared with other sources of AMR in different watersheds, which can inform decision-making and risk assessments. Research is also needed to track and understand how ARGs are propagated in wastewater microbial communities and how ARGs can be transferred from wastewater sources to environmental bacteria and potentially on to pathogenic microorganisms in the environment.

C. Authority and Regulations

The authorities for this RFA and resulting awards are contained in the Clean Water Act, 33 U.S.C. 1254, Section 104(b)(3), the Safe Drinking Water Act, 42 U.S.C. 300j-1, Section 1442, and the Consolidated Appropriations Act, 2023, Public Law 117-328.

For research with an international aspect, the above statutes are supplemented, as appropriate, by the National Environmental Policy Act, Section 102(2)(F).

Note that a project’s focus is to consist of activities within the statutory terms of EPA’s financial assistance authorities; specifically, the statute(s) listed above. Further note applications dealing with any aspect of or related to hydraulic fracking will not be funded by EPA through this program.

Additional applicable regulations include: 2 CFR Part 200, 2 CFR Part 1500, and 40 CFR Part 40 (Research and Demonstration Grants).

D. Specific Research Areas of Interest/Expected Outputs and Outcomes Note to applicant: The term “output” means an environmental activity, effort, and/or associated work products related to an environmental goal or objective, that will be produced or provided over a period of time or by a specified date. The term “outcome” means the result, effect, or consequence that will occur from carrying out an environmental program or activity that is related to an environmental or programmatic goal or objective.

The activities to be funded under this solicitation support EPA’s FY 2022-2026 Strategic Plan (https://www.epa.gov/planandbudget/strategicplan). Awards made under this solicitation will support Goal 5: Ensure Clean and Safe Water for All Communities, Objective 5.1: Ensure Safe Drinking Water and Reliable Water Infrastructure and Objective 5.2: Protect and Restore Waterbodies and Watersheds, of the Plan. All applications must be for projects that support the goal(s) and objective(s) identified above. Awards made under this announcement will further the https://www.epa.gov/planandbudget/strategicplan goals of the Consolidated Research/Training/Fellowships program by furthering EPA’s priorities ensuring clean and safe water for safe and sustainable water resources by promoting high-priority water quality and availability research. The agency is seeking applications proposing innovative research to address knowledge gaps on the occurrence, fate and transport, and persistence of antimicrobial resistant organisms and genes found in municipal wastewater effluent and biosolids.

EPA also requires that grant applicants adequately describe environmental outputs and outcomes to be achieved under assistance agreements (see EPA Order 5700.7A1, Environmental Results under Assistance Agreements, https://www.epa.gov/grants/epa-order-57007a1-epas-policy-environmental-results-under-epa-assistance-agreements). Applicants must include specific statements describing the environmental results of the proposed project in terms of well-defined outputs and, to the maximum extent practicable, well-defined outcomes that will demonstrate how the project will contribute to the goal(s) and objective(s) described above.

The Agency is soliciting research that will address knowledge gaps on the occurrence, fate and transport, and persistence of antimicrobial resistant organisms and genes found in municipal wastewater effluent and biosolids. Note that the research should be national in scope; it should also focus on water quality (by looking at the impact of ARB and ARGs in wastewater effluent on receiving waters used for drinking water, recreation, support of ecosystems, and other purposes) and water availability (to the extent that improving water quality increases the amount of safe drinking water to the public). The proposed research will provide new information needed to determine the removal efficiency of ARB and ARGs in wastewater treatment plants, national estimates of AMR discharges to U.S. surface waters via treated wastewater, and an increased ability to understand and identify potential public health and ecosystem risks from ARB and ARGs in receiving waters. The proposed research should support or contribute to the ability to conduct risk assessments and eventually estimate risk of infection for drinking water, recreational water, and occupational exposures. Additionally, it should facilitate comparison of AMR impacts from treated wastewater effluent and biosolids compared to other non-municipal wastewater sources (e.g., animal agriculture, hospital wastes, other industrial sources, and untreated wastewater discharges).

Given the lack of standardized methods, applicants should also clearly describe selected AMR targets and methods proposed, as well as how they best characterize AMR and address the proposed study objectives. Current work to standardize methods in the literature are Liguori et al., (2022) and The Water Research Foundation (2023) which include frameworks for AMR monitoring of wastewater, recycled water, and surface water. Franklin et al. (2020), provides an overview of common contemporary molecular methods available for analyzing AMR in surface waters and associated rationales for selection of methods dependent on the study outcome.

Specifically, with respect to molecular methods, the Minimum Information for Publication of Quantitative Real-Time Polymerase Chain Reaction Experiments (MIQE) guidelines include minimum information necessary for evaluating quantitative polymerase chain reaction (qPCR) experiments and directions for researchers to develop and publish research using qPCR methods.

https://www.epa.gov/grants/epa-order-57007a1-epas-policy-environmental-results-under-epa-assistance-agreements https://www.epa.gov/grants/epa-order-57007a1-epas-policy-environmental-results-under-epa-assistance-agreements

The proposed research should account for these and other sources of methods standardization to ensure that project data and results are transferrable and generalizable given their specific objective. Last, research should be planned in the context of existing literature, leveraging existing data sources to the extent possible, and focusing data collection on outstanding needs.

There are two distinct areas of research covered by this solicitation. EPA encourages applicants to focus on only one research area. However, applications that address more than one research area will not be deemed ineligible but will not necessarily be rated more highly than those that address just one. In addition, applications that are not national in scope may not be rated as highly as those that are. The proposed research project should be as responsive as possible to as many of the research questions listed under the selected research area; however, applicants are not required to respond to all the research questions or limit the research scope to these questions. Applications should clearly indicate which research area is being addressed.

Research Areas Research Area 1: Understanding the selection and removal of antimicrobial resistant genes and bacteria throughout wastewater and biosolids treatment trains. The goal of this research area is to better understand the contribution of different wastewater treatment processes on the potential proliferation or removal efficiency of ARB and ARGs. Emphasis should be placed on unit processes and treatment trains that are broadly representative of U.S. wastewater treatment plants (WWTPs), including both large and small systems discharging to surface water, groundwater, and waterbodies with varying designated uses (e.g., water supply, primary contact recreation, aquatic life, and water reuse). Results should fill knowledge gaps in the understanding of the fate of AMR in different conventional and advanced municipal wastewater treatment processes. Additionally, research should support the development of national-scale estimates of ARB and ARG discharges from WWTP effluents, and support comparison to other non-municipal wastewater sources. These estimates should be generated such that they are compatible with relevant monitoring programs such as the National Antimicrobial Resistance Monitoring System (NARMS) and National Wastewater Surveillance System (NWSS). Note that this research area also requests information to better understand how sludge treatment processes effect ARB and ARGs found in biosolids.

The following are suggested research questions to be considered. Applicants are encouraged to consider other significant questions that need to be investigated to meet the objectives of this research area:

• What are the key operational and site-specific parameters impacting AMR selection, growth, and removal in conventional and advanced treatment processes and sewage sludge treatment processes? In addition to operational and site-specific parameters, this includes aspects such as physiochemical wastewater parameters, antibiotic concentrations, operational conditions, dry or wet weather flow, as well as mechanisms responsible for attenuation that impact AMR.

https://www.fda.gov/animal-veterinary/antimicrobial-resistance/national-antimicrobial-resistance-monitoring-system https://www.fda.gov/animal-veterinary/antimicrobial-resistance/national-antimicrobial-resistance-monitoring-system https://www.cdc.gov/nwss/wastewater-surveillance/index.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.gov%2Fhealthywater%2Fsurveillance%2Fwastewater-surveillance%2Fwastewater-surveillance.html

• What key knowledge gaps need to be addressed on the removal of ARB and ARGs by oxidation and disinfection processes and on the impact of typical doses used on AMR of final treated wastewater?

• How do combinations of conventional and advanced treatment processes impact the removal of AMR and what current knowledge gaps need to be addressed?

o How can ARB and ARG levels be controlled within treatment processes?

• How do WWTP characteristics such as size of system, types of treatment processes, location, and treatment train configuration affect ARB, ARG, and AMR residue levels in WWTP effluent?

o How effective is conventional and advanced treatment (including chlorination, UV treatment, ozonation, and advanced oxidation processes) in removing ARB and ARGs?

o What is the occurrence, mechanism, and timing of genetic mobilization of ARGs between microbial communities in wastewater reclamation facilities?

o What are the differences between wastewater systems of the same size using different treatment trains?

• What are the levels of ARB and ARGs in influent as compared to levels in effluent?

• How do WWTPs vary nationally regarding removal efficiencies of ARB and ARGs?

o What are the impacts of watershed geography, temperature, and socioeconomic aspects on ARB and ARG levels in effluent?

o What are the differences in ARB and ARGs in effluent across different treatment trains?

• What is the impact from different influent wastewaters such as municipal dominated vs systems including commercial and/or industrial sources, and hospitals on ARB, ARG, and antimicrobial levels in effluent?

o How does this impact loadings and selection or removal within the individual treatment processes?

• What is the impact of different biosolids treatment processes such as anaerobic digestion, lime stabilization, heat drying, and composting on AMR?

o How do different operational factors affect AMR in biosolids?

o How well do these biosolids processes impact antimicrobial residues?

Research Area 2: Understanding the environmental burden and public health impact of antimicrobial resistant genes and bacteria from different municipal wastewater sources on downstream applications and from biosolids use on the environment. Multiple wastewater sources of AMR, including municipal effluents, land-applied biosolids, septic systems, and untreated sewage overflows may contribute to AMR levels in receiving waters that are used for recreation, aquatic life, ecosystem services, and planned and de facto reuse. New information is needed to characterize the different wastewater sources so that the relative contributions can be determined and compared to other sources such as agriculture. Moreover, this research area includes the development and demonstration of risk assessment models (e.g., of acute human exposure risks, ecological impacts, and the development of novel resistances among pathogens).

Study design should therefore facilitate the use of generated data in risk assessments and relative source contribution studies.

The following are suggested research questions to be considered. Applicants are encouraged to consider other significant questions that need to be investigated to meet the objectives of this research area:

• What is the relative significance of wastewater effluent as a source of ARB and ARGs in receiving waters compared to other non-wastewater sources such as animal agriculture, animal husbandry, hospital sources, direct industrial sources, etc.?

o What is the relative contribution of ARB and ARGs from wastewater effluent (secondary treated and disinfected), combined sewer overflows (CSOs), septic systems, and small systems such as lagoon systems, to receiving surface waters?

o What is the impact from gene transfer in the environment, compared to other non-wastewater related pathways such as natural development, animal husbandry, animal agriculture, and others?

• How can effluent measurements be used to estimate human health risks associated with water reuse (potable, non-potable, de facto), primary contact recreation, and other potential exposures?

• What is the significance of AMR from land applied biosolids on human and animal health and the environment?

o What is the role of biosolids (Class A and Class B) on the spread of AMR to the environment?

o What is the impact on food crops? On domesticated and wild animals?

o How do land applied biosolids transfer ARGs to human pathogens?

• What are appropriate mitigation and management practices that reduce the AMR risk from land applied biosolids?

• What are the appropriate frameworks for quantifying risk related to AMR in treated wastewater discharge, water reuse, and biosolids and land application?

Expected Outputs/Outcomes

Expected Outputs: Some examples of desirable outputs are listed below:

• Bench, pilot, or full-scale testing at wastewater facilities and biosolids land application sites.

• Knowledge development on risk from effluent discharge and biosolids land application and other beneficial uses.

• Development of appropriate frameworks and methodologies for quantifying risk related to AMR in treated wastewater discharge, water reuse, and biosolids (and biosolids products) land application and beneficial use.

• Assessment of the significance of the wastewater discharge sources and biosolids use on AMR proliferation in the environment in comparison to other non-wastewater sources in different watersheds.

• Research and testing reports and peer reviewed publications pertaining to the research areas listed above.

• Workshops and webinars to disseminate information to states and utilities.

• Guidance documents for utilities and states including targets and ways to integrate with other networks.

o Guidance to mitigate the sources of ARB and ARGs, particularly those that may persist and/or accumulate in the environment.

• Risk assessments along with development of risk models and source apportionment studies.

• Data on wastewater effluent quality as impacted by various processes and geographical variation.

• Survey frameworks for the longitudinal monitoring of AMR in wastewater effluents.

• Guidance on effective treatment processes and operational configurations for the management of ARB and ARGs in wastewater.

Expected Outcomes:

• Better understanding of the nature, extent, selection, and removal of ARB and ARGs found in municipal wastewater effluent and biosolids as well as the conditions associated with their occurrence and growth.

• Improved understanding of the environmental burden and public health impact of ARB and ARGs from different municipal wastewater sources on downstream applications and from biosolids use on the environment.

• Better planning and increased ability for States, municipalities, and wastewater treatment facilities to identify and manage human health and ecosystem risks from ARB and ARGs.

E. References

CDC, Centers for Disease Control and Prevention (2019) Antibiotic Resistance Threats in the United States. Retrieved from: https://www.cdc.gov/drugresistance/pdf/threats-report/2019-ar-threats-report-508.pdf.

CDC, Centers for Disease Control and Prevention. One Health Basics. Retrieved from:

https://www.cdc.gov/onehealth/basics/index.html.

Christou, A., Agüera, A., Bayona, J. M., Cytryn, E., Fotopoulos, V., Lambropoulou.,…Fatta- Kassinos, D. (2017). The potential implications of reclaimed wastewater reuse for irrigation on the agricultural environment: the knowns and unknowns of the fate of antibiotics and antibiotic resistant bacteria and resistance genes–a review. Water research, 123, 448-467.

https://www.cdc.gov/drugresistance/pdf/threats-report/2019-ar-threats-report-508.pdf https://www.cdc.gov/drugresistance/pdf/threats-report/2019-ar-threats-report-508.pdf https://www.cdc.gov/onehealth/basics/index.html

Coleman, B. L., Salvadori, M. I., McGeer, A. J., Sibley, K. A., Neumann, N. F., Bondy.,…Louie, M. (2012). The role of drinking water in the transmission of antimicrobial-resistant E. coli.

Epidemiology & Infection, 140(4), 633-642.

Damashek, J., Westrich, J. R., McDonald, J. M. B., Teachey, M. E., Jackson, C. R., Frye, J. G., ...Ottesen, E. A. (2022). Non-point source fecal contamination from aging wastewater infrastructure is a primary driver of antibiotic resistance in surface waters. Water Research, 222, 118853.

Eramo, A., Delos Reyes, H., & Fahrenfeld, N. L. (2017). Partitioning of antibiotic resistance genes and fecal indicators varies intra and inter-storm during combined sewer overflows.

Frontiers in microbiology, 8, 2024.

Franklin, A. M., Brinkman, N. E., Jahne, M. A., & Keely, S. P. (2021). Twenty-first century molecular methods for analyzing antimicrobial resistance in surface waters to support One Health assessments. Journal of microbiological methods, 184, 106174.

Hölzel, C. S., Tetens, J. L., & Schwaiger, K. (2018). Unraveling the role of vegetables in spreading antimicrobial-resistant bacteria: a need for quantitative risk assessment. Foodborne pathogens and disease, 15(11), 671-688.

Law, A., Solano, O., Brown, C. J., Hunter, S. S., Fagnan, M., Top, E. M., & Stalder, T. (2021).

Biosolids as a source of antibiotic resistance plasmids for commensal and pathogenic bacteria.

Frontiers in microbiology, 709.

Leonard, A. F., Zhang, L., Balfour, A. J., Garside, R., Hawkey, P. M., Murray, A. K., … Gaze, W. H. (2018). Exposure to and colonisation by antibiotic-resistant E. coli in UK coastal water users: Environmental surveillance, exposure assessment, and epidemiological study (Beach Bum Survey). Environment international, 114, 326-333.

Liguori, K., Keenum, I., Davis, B. C., Calarco, J., Milligan, E., Harwood, V. J., & Pruden, A.

(2022). Antimicrobial Resistance monitoring of water environments: A framework for standardized methods and quality control. Environmental science & technology, 56(13), 9149- 9160.

National Academies of Sciences, Engineering, and Medicine. (2022). Combating Antimicrobial Resistance and Protecting the Miracle of Modern Medicine. The National Academies Press.

Retrieved from: https://nap.nationalacademies.org/catalog/26350/combating-antimicrobial-resistance-and-protecting-the-miracle-of-modern-medicine.

Sanganyado, E., & Gwenzi, W. (2019). Antibiotic resistance in drinking water systems:

Occurrence, removal, and human health risks. Science of the Total Environment, 669, 785-797.

https://nap.nationalacademies.org/catalog/26350/combating-antimicrobial-resistance-and-protecting-the-miracle-of-modern-medicine https://nap.nationalacademies.org/catalog/26350/combating-antimicrobial-resistance-and-protecting-the-miracle-of-modern-medicine

Søraas, A., Sundsfjord, A., Sandven, I., Brunborg, C., & Jenum, P. A. (2013). Risk factors for community-acquired urinary tract infections caused by ESBL-producing enterobacteriaceae–a case–control study in a low prevalence country. PloS one, 8(7), e69581.

The Water Research Foundation (2018). Occurrence, proliferation, and persistence of antibiotics and antibiotic resistance during wastewater treatment. Retrieved from:

https://www.waterrf.org/research/projects/occurrence-proliferation-and-persistence-antibiotics-and-antibiotic-resistance-0.

The Water Research Foundation (2023). Standardizing Methods with QA/QC Standards for Investigating the Occurrence and Removal of Antibiotic Resistant Bacteria/Antibiotic Resistance Genes (ARB/ARGs) in Surface Water, Wastewater, and Recycled Water. Retrieved from:

https://www.waterrf.org/research/projects/standardizing-methods-qaqc-standards-investigating-occurrence-and-removal.

Tyrrell, C., Burgess, C. M., Brennan, F. P., & Walsh, F. (2019). Antibiotic resistance in grass and soil. Biochemical Society Transactions, 47(1), 477-486.

Wengenroth, L., Berglund, F., Blaak, H., Chifiriuc, M. C., Flach, C. F., Pircalabioru, G. G., ... & Schmitt, H. (2021). Antibiotic resistance in wastewater treatment plants and transmission risks for employees and residents: The concept of the AWARE study. Antibiotics, 10(5), 478.

F. Special Requirements

It is EPA Policy to ensure that the results of EPA-funded extramural scientific research are accessible to the public to the greatest extent feasible consistent with applicable law; policies and Orders; the Agency’s mission; resource constraints; and U.S. national, homeland and economic security. This entails maximizing, at no charge, access by the public to peer-reviewed, scientific research journal publications or associated author manuscripts, and their underlying digital research data, created in whole or in part with EPA funds, while protecting personal privacy;

recognizing proprietary interests, confidential business information, and intellectual property rights; and avoiding significant negative impact on intellectual property rights, innovation, and U.S. competitiveness.

Applications submitted under this announcement shall include a Scientific Data Management Plan (SDMP) that addresses public access to EPA-funded scientific research data. See the SDMP clause in Section IV for details on the content of an SDMP. Applicants will also be asked to provide past performance information on whether journal publications or associated author manuscripts, and the associated underlying scientific research data and metadata, under prior assistance agreements were made publicly accessible. These items will be evaluated prior to award.

https://www.waterrf.org/research/projects/occurrence-proliferation-and-persistence-antibiotics-and-antibiotic-resistance-0 https://www.waterrf.org/research/projects/occurrence-proliferation-and-persistence-antibiotics-and-antibiotic-resistance-0 https://www.waterrf.org/research/projects/standardizing-methods-qaqc-standards-investigating-occurrence-and-removal https://www.waterrf.org/research/projects/standardizing-methods-qaqc-standards-investigating-occurrence-and-removal

Reasonable, necessary and allocable costs for data management and public access may be included in extramural research applications and detailed in the budget justification described in Section IV.

Agency policy and ethical considerations prevent EPA technical staff and managers from providing applicants with information that may create an unfair competitive advantage.

Consequently, EPA employees will not review, comment, advise, and/or provide technical assistance to applicants preparing applications in response to EPA RFAs. EPA employees cannot endorse any particular application.

Multiple Investigator applications may be submitted as: (1) a single Lead Principal Investigator (PI) application with Co-PI(s) or (2) a Multiple PI application (with a single Contact PI). If you choose to submit a Multiple PI application, you must follow the specific instructions provided in Sections IV and V of this RFA. For further information, please see the EPA Implementation Plan for Policy on Multiple Principal Investigators (https://www.epa.gov/research-grants/research-grants-guidance-and-policies).

This solicitation provides the opportunity for the submission of applications for projects that may involve human subjects research. All applications must include a Human Subjects Research Statement (HSRS; described in Section IV.C.5.iii.c of this solicitation). If the project involves human subjects research, it will be subject to an additional level of review prior to funding decisions being made as described in Sections V.D and V.F of this solicitation.

These awards may involve the collection of “Geospatial Information,” which includes information that identifies the geographic location and characteristics of natural or constructed features or boundaries on the Earth or applications, tools, and hardware associated with the generation, maintenance, or distribution of such information. This information may be derived from, among other things, a Geographic Positioning System (GPS), remote sensing, mapping, charting, and surveying technologies, or statistical data.

G. Additional Provisions for Applicants Incorporated into the Solicitation

Additional provisions that apply to sections III, IV, V, and VI of this solicitation and/or awards made under this solicitation, can be found at EPA Solicitation Clauses. These provisions are important for applying to this solicitation and applicants must review them when preparing applications for this solicitation. If you are unable to access these provisions electronically at the website above, please contact the EPA point of contact listed in this solicitation (usually in Section VII) to obtain the provisions.

II. AWARD INFORMATION

It is anticipated that a total of approximately $9,500,000 will be awarded under this announcement, depending on the availability of funds, quality of applications received, and other https://www.epa.gov/research-grants/research-grants-guidance-and-policies https://www.epa.gov/research-grants/research-grants-guidance-and-policies https://www.epa.gov/grants/epa-solicitation-clauses applicable considerations. The EPA anticipates funding approximately 4 awards under this RFA.

Requests for amounts in excess of a total of $2,375,000 in federal funds per award, including direct and indirect costs, will not be considered. In addition, a minimum 25% non-federal cost share/match of the federal funds awarded, which may include in-kind contributions (see Section III.B. for more details), is required. For example, if an applicant requests $2,375,000 in EPA funds, a minimum of $593,750 must be included. Including matching, total project costs can exceed $2,968,750 (if the applicant proposes more than the minimum required non-federal cost share/match), however, the federally-funded portion of the budget must not exceed $2,375,000.

Applications which do not include the minimum 25% non-federal cost share/match will not be considered. The total project period requested in an application submitted for this RFA may not exceed 3 years.

The EPA reserves the right to reject all applications and make no awards, or make fewer awards than anticipated, under this RFA. The EPA reserves the right to make additional awards under this announcement, consistent with Agency policy, if additional funding becomes available after the original selections are made. Any additional selections for awards will be made no later than six months after the original selection decisions.

In appropriate circumstances, EPA reserves the right to partially fund applications by funding discrete portions or phases of proposed projects. If EPA decides to partially fund an application, it will do so in a manner that does not prejudice any applicants or affect the basis upon which the application, or portion thereof, was evaluated and selected for award, and therefore maintains the integrity of the competition and selection process. Awards may be fully or incrementally funded, as appropriate, based on funding availability, satisfactory performance, and other applicable considerations.

EPA intends to award only grants under this announcement.

Under a grant, EPA scientists and engineers are not permitted to be substantially involved in the execution of the research. However, EPA encourages interaction between its own laboratory scientists and grant Principal Investigators after the award of an EPA grant for the sole purpose of exchanging information in research areas of common interest that may add value to their respective research activities. This interaction must be incidental rather than substantial to achieving the goals of the research under a grant. Interaction that is “incidental” does not involve resource commitments by EPA.

III. ELIGIBILITY INFORMATION

Note: Additional provisions that apply to this section can be found at EPA Solicitation Clauses.

A. Eligible Applicants

This solicitation is available to public and private nonprofit institutions and public and private universities and colleges located in the United States and its territories or possessions. Foreign entities, U.S. States, territories and possessions, the District of Columbia, State and local government departments, and Federally Recognized Indian Tribal Governments of the U.S., are not eligible to apply under this RFA. Profit-making firms and individuals are not eligible to receive assistance agreements from the EPA under this program.

Consistent with the definition of Nonprofit organization at 2 CFR § 200.1, the term nonprofit organization means any corporation, trust, association, cooperative, or other organization that is operated mainly for scientific, educational, service, charitable, or similar purpose in the public interest and is not organized primarily for profit; and uses net proceeds to maintain, improve, or expand the operation of the organization. Note that 2 CFR § 200.1 specifically excludes Institutions of Higher Education from the definition of non-profit organization because they are separately defined in the regulation. While not considered to be a nonprofit organization(s) as defined by 2 CFR § 200.1, public or nonprofit Institutions of Higher Education are, nevertheless, eligible to submit applications under this RFA. Hospitals that meet the definition of nonprofit at 2 CFR § 200.1 are also eligible to apply as nonprofits. Hospitals operated by state, tribal, or local governments or that are instrumentalities of the unit of government depending on the applicable law are not eligible to apply. For-profit colleges, universities, trade schools, and hospitals are ineligible.

Nonprofit organizations that are not exempt from taxation under section 501 of the Internal Revenue Code must submit other forms of documentation of nonprofit status; such as certificates of incorporation as nonprofit under state or tribal law. Nonprofit organizations exempt from taxation under section 501(c)(4) of the Internal Revenue Code that lobby are not eligible for EPA funding as provided in the Lobbying Disclosure Act, 2 U.S.C. 1611.

National laboratories funded by Federal Agencies (Federally-Funded Research and Development Centers, “FFRDCs”) may not apply. FFRDC employees may cooperate or collaborate with eligible applicants within the limits imposed by applicable legislation and regulations. They may participate in planning, conducting, and analyzing the research directed by the applicant, but may not direct projects on behalf of the applicant organization. An award recipient may provide funds through its assistance agreement from the EPA to an FFRDC for research personnel, supplies, equipment, and other expenses directly related to the research.

Federal Agencies may not apply. Federal employees are not eligible to serve in a principal leadership role on an assistance agreement. Federal employees may not receive salaries or augment their Agency’s appropriations through awards made under this program unless authorized by law to receive such funding.

The applicant institution may enter into an agreement with a Federal Agency to purchase or utilize unique supplies or services unavailable in the private sector to the extent authorized by law. Examples are purchase of satellite data, chemical reference standards, analyses, or use of instrumentation or other facilities not available elsewhere. A written justification for federal involvement must be included in the application. In addition, an appropriate form of assurance that documents the commitment, such as a letter of intent from the Federal Agency involved, should be included.

Potential applicants who are uncertain of their eligibility should contact Ron Josephson in ORD, phone: 202-564-7823, email: josephson.ron@epa.gov.

B. Cost sharing

Each applicant must contribute a minimum non-federal cost share/match of 25% of the federal funds awarded. This is equivalent at a minimum to 20% of the total project costs.

For example, if an applicant requests $2,375,000 in EPA funds, a minimum of $593,750 must be included. Including matching, total project costs can exceed $2,968,750 (if the applicant proposes more than the minimum required non-federal cost share/match), however, the federally-funded portion of the budget must not exceed $2,375,000.

If the applicant is successful, the resultant assistance agreement will display cost share as a percentage of total project costs. Cost share may include in-kind contributions. In order to be eligible for funding consideration, applicants must demonstrate in their applications how they will meet the required minimum 25% cost share/match in accordance with 2 CFR § 200.306.

The cost share/match may be provided in cash or can come from in-kind contributions, such as the use of volunteers and/or donated time, equipment, etc., subject to the regulations governing matching fund requirements at 2 CFR § 200.306. Cost share/matching funds are considered grant funds and are included in the total award amount.

All contributions, including cash and third party in-kind, shall be accepted as cost sharing or matching when such contributions meet all of the following criteria: (1) Are verifiable from the non-Federal entity's records; (2) Are not included as contributions for any other Federal award;

(3) Are necessary and reasonable for proper and…