Source: UNIVERSITY OF KENTUCKY submitted to NRP
EVALUATION OF NANOPESTICIDE FATE AND TRANSPORT AND BIOGEOCHEMICAL IMPLICATIONS IN AGROECOSYSTEMS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1025426
Grant No.
2021-67021-34368
Cumulative Award Amt.
$479,763.00
Proposal No.
2020-04116
Multistate No.
(N/A)
Project Start Date
Feb 15, 2021
Project End Date
Feb 14, 2026
Grant Year
2021
Program Code
[A1511]- Agriculture Systems and Technology: Nanotechnology for Agricultural and Food Systems
Recipient Organization
UNIVERSITY OF KENTUCKY
500 S LIMESTONE 109 KINKEAD HALL
LEXINGTON,KY 40526-0001
Performing Department
Biosystems and Ag Engineering
Non Technical Summary
Recent discoveries of nanopesticide influence on nitrogen removal processesprovides an urgency to understand the impacts of these nanopesticides in realistic field settings. The persistence, transport, and functional response to the nitrogen cycle fromto these engineered nanomaterials is only just being explored now at the laboratory setting. Therefore, the overarching goals of the proposed work are to i) evaluate nanopesticide impact on the nitrogen cycle, ii) quantify their persistence, and iii) identify transport scenarios of nanopesticide usage in two distinct agricultural settings (Southeast and Northwest). Potential nitrogen cycling will be assessed using microcosms of field and riparian soils. Persistence and transport pathways will be assessed using coupled mesocosms and field scale studies to ensure findings are comparable in realistic settings. Lastly, a nanopesticide fate and transport model for both nanopesticide runoff and leaching will be completed using a variety of fate and transport models. Findings are anticipated to provide novel insight to nanopesticide detection, transport, fate, degradation, and characterization. Results will guide growers and the nanopesticide market on innovative methods for improving and/or maintaining crop productivity while preserving and protecting environmental/soil health and sustainability of our natural resource base. Expected deliverables include: i) realistic recommendations for minimizing environmental impacts of nanopesticides, ii) fate and transport methodology of nanopesticides that will be transferable and scalable to other questions in agricultural fields and downstream best management practices, iii) predictive modeling capabilities for nanopesticide fate and transport, and iv) undergraduate and graduate training.
Animal Health Component
50%
Research Effort Categories
Basic
25%
Applied
50%
Developmental
25%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1020210202050%
1020210200025%
1020210100025%
Knowledge Area
102 - Soil, Plant, Water, Nutrient Relationships;

Subject Of Investigation
0210 - Water resources;

Field Of Science
2020 - Engineering; 1000 - Biochemistry and biophysics; 2000 - Chemistry;
Goals / Objectives
Theoverall goalsof the proposed work are to i) evaluate nanopesticide impact on the nitrogen cycle, ii) quantify their persistence, and iii) identify transport scenarios of nanopesticide usage in two distinct agricultural settings (Southeast and Northwest).The objectives will be supported by experiments that include quantitative measurements of mixture effects between a commercially available nano-Cu fungicide (i.e.,Cu(OH)2)and environmentally relevant concentrations of selected neonicotinoids and inorganic nitrogen (N) fertilizer likely to occur in runoff from agricultural fields in the Southeast and Northwest.Our specific researchobjectivesare:Complete coupled microcosm/mesocosm scale experiments to assess microbial alterations and neonicotinoidinsecticide degradation based on varying nanopesticide application and recurrance rates;Conduct a 2 year field study to assess nanopesticide pesistance, fate, and transport;Create kinectic models from bench, mesocom,and field scale experiments and incorporate findings into HYDRUS modeling software to predict fate and transport of nanopesticides into the vadose zone;
Project Methods
Research Objective 1 (RO1):Complete coupled microcosm/mesocosm scale experiments to assess microbial alterations and neonicotinoid insecticidedegradation based on varying nanopesticide application and recurrance rates;Microcosm Experimental Design and Methods (RO1a).RO1a will utilize microcosms to assess nanopesticide treatments on nutrient transformation, leaching, and greenhouse gas emissions from four soil sources: a silt loam cropland soil from Kentucky (Southeastcrop soil), a riparian wetland sediment collected in close proximity to the Kentucky crop soil (Southeast wetland sediment), a cropland soil from Washington (Northwest crop soil), and riparian wetland sediment collected in close proximity to the Washington crop soil (Northwest wetland sediment). Each microcosm will be constructed using 20-mL syringes fitted with glass fiber filters in the tip.Each syringe will be filled with 15 mL of soil or sediment with 5 mL of artificial runoff water (based on surface water collected from the field sites) applied weekly.During the study, cropland soils will be allowed to immediately leach applied runoff water, while riparian soils will be held at saturation and only drained at the end of the week, immediately before a fresh runoff application. Leachate composition, greenhouse gas fluxes, and microbial denitrification and nitrification will be assessed initially, at 5 weeks, and at 10 weeks.Mesocosm Experimental Design and Methods (RO1b).Mesocosm experiments will be completed in research greenhouses located at the University of Kentuckyand Western Washington University to assess nanopesticideimplications on downstream best management practices. In summary, each greenhouse will be outfitted with eighteen mesocosms. To create the mesocosms, eighteen 100-gallon storage tanks will contain 12" to 24" ofsoils from the same locations of the microcosm experiments. Twelve of the Kentucky riparian soils will have rooted in native herbaceous plants commonly found in regional riparian wetlands (i.e.,Carex comosa) in 24" of sediment. Similarly, twelve of the Washington riparian soils will utilize a native sedge plant (Carex leptopoda) in order to have a more direct comparison between soil types and plant species. Plants will be preestablished during year one of the project and the mesocosm experiments will be conducted in year 2.Four experiments will be conducted with mixtures of nitrogen, nanopesticides, and complex interactions are expected to impact the dominant nutrient kinetic rates and degradation products andrates of the pesticides in the mesocosms dependent on the time of day, UV exposure, and plant growth.Baselinenanopesticideand N species concentrations will be collected prior to enriching systems. Bromide will be added to the wetland mesocosms to determine evapotranspiration and adjust analyte concentrations. Temperature and light exposure will be measured using an Onset datalogger in the greenhouses in each of the mesocosm soil surfaces. Plant samples will be taken prior to and after the completion of each experiment to conduct a N mass balanceto estimate differences between denitrification and plant uptake. Findings will be compared to results from the acetylene block observations in RO1a. PCommon transformation pesticide byproducts along with parent pesticides will be measured every 1 to 3 days in the soil and water, while plant uptake of the pesticides will be assessed prior to pesticide application, 30 days following pesticide application, and at the completion of the experiment.Kinetic Removal Models.Kinetic models will be created to provide tools for stakeholders to predict the NO3-N removal capacity based on removal pathways in the two wetland designs in varying water chemistry scenarios. Concentrations of NO3-N will be fit to a first order decay response model. However, other response models (zero order, Michaelis-Menton, efficiency loss, etc.) will be evaluated if removal rates do not adequately fit the first order removal model utilizing methods.Research Objective 2:Conduct a 2 year field study to assess nanopesticide pesistance, fate, and transport;Field Sites Experimental Design.Field plots will be established at the University of Kentucky in mid-April and monitored through mid-September of years 2 and 3 of the study. Dimensions of the plots will be approximately 3.6 m wide by 7 m in length using 20 cm-wide sheet metal frames driven approximately 10 cm into the soil. A total of 4 treatments (1. control-no pesticide application, 2.Nano-Cu fungicide application, 3.nano-imidaclopridapplication, and 4.Nano-Cu fungicide and nano-imidacloprid application) will be assessed in replicates of three.Field Site Methods.Soil nutrient content will be measured, and fertilizer will be applied to the cropland field plots at an agronomic N rate for vineyards. Nanopesticide applications will be applied to nine of the twelve plots at the beginning of the growing season. Nano-Cu fungicide will be applied as recommended by the pesticide label to six of the twelve plots and will be reapplied based on the pesticide label throughout the growing season. Similarly, nanopesticideswill be applied at the beginning of the growingas recommended by the pesticide label to six of the twelve plots and will be reapplied based on the pesticide label throughout the growing season.Bromide will be applied to all plots as a conservative tracer to be utilized in the fate and transport model (RO3). Transport of thenanopesticideswill be monitored after naturally occurring rainfall over a 20 to 30 week period during the growing season (April through October). Soil samples from the plots will be collected before and after pesticide applications to establish any baseline concentrations ofnano-Cufungicide, N, and neonicotinoids.Runoff samples will be collected following each natural rainfall event to quantify of potentialnanopesticides being transported off the field plots. Lastly, soil samples will be collected monthly during the two year experiment along three transects (upland, middle, base) at three depths (0-5 cm, 5-15 cm, and 15-30 cm) in each plot to determine the vertical transport ofthe nanopesticidesinto the vadose zone.Research Objective 3:Create kinetic models from bench, mesocosm, and field scale experiments and incorporate findings into HYDRUS modeling software to predict fate and transport of nanopesticides into the vadose zone;We will explorethe feasibility of four vadose modeling frameworks to investigate their potential for fate and transport of nanopesticides in agroecosystem vadose zones: HYDRUS-2D, VLEACH, GLEAMS, and LEACHM. The following models are proposed for investigation of fate and transport of nanopesticides given their usage for traditionally sized pesticides. Further, the various models requiring varying input parameters and the investigation in this study will provide a holistic sensitivity assessment for the necessary parameters required to predict fate and transport of nano pesticides into the vadose zone in contrast to the current models traditionally sized pesticides. We will utilize both the data collected from R1 and R2 to estimate KOCand half-life values of the nanopesticides using inverse modeling. However, we will explore alternative models (e.g., GLEAMS, VLEACH, LEACHM) as well and complete a sensitivity analysis for various variables (e.g., hydraulic conductivity, macropore space, nitrogen content, carbon content, solute dynamics, preferential flow; plants) to determine which model is the best predictor for estimating leaching of nanopesticides at the microcosm, mesocosm, and field scales.

Progress 02/15/24 to 02/14/25

Outputs
Target Audience:During the 2023-2024 year we further expanded our outreach programs to include 12 high schools and over 600 high school students and teachers. Updates to the advisory was completed by Dr. Messer to representatives from Kentucky Extension Program and Fayette County Public Schools. Ten trainings were completed by Dr. Messer for high school educators who plan to share activities with their classrooms over the next 12 months. Pumpkins grown as part of the project were donatedonce again to local schools for classroom Halloween projects and local farmer's markets in addition with sharing information regarding the research project. We have presented findings at nine conferences (Gordan Research Conference (Manchester, NH), Kentucky Water Research Institute Annual Meeting (Lexington, KY), 2024 ASA, CSSA, and SSSA International Annual Meeting (San Antonio, TX), SouthOn+ Southeast Regional Research Symposium (Lexington, KY), 2024 USDA NIFA Annual Meeting (Knoxville, TN), American Ecological Engineering Conference (Blacksburg, Virginia), 2025 American Society of Agricultural and Biological Engineering Conference (Anaheim, California), 2024 Pacific Northwest Society of EnvironmentalToxicology and Chemistry meeting (Troutdale, OR), North American Society of Environmental Toxicology and Chemistry Conference (Fort Worth, TX)). Overall, we reached over 4,000 people throughout outreach events with ages ranging from pre-K to over 60 years in age. Further, 4 undergraduate and 4 graduate students haveworked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1-3 during the 2023-2024 yearand one graduate student completed her MS thesis as part of this project in 2024. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?4 undergraduate and 4 graduate students have worked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1, 2, and 3 in2023-2024. Students were trained on laboratory analysis, experimental setup, sampling protocol, and all MS students completed their MS proposals related to this project. How have the results been disseminated to communities of interest?Overall, we reachedapproximately over 4,000people through outreachevents with ages ranging from Pre-K to over 60 years in age. This was completed by presenting findings at nine conferences, ten high schools (many were Title 1), and the Pre-K outreach programmesoWheels. Further, we have a publication under review and a master's thesis that would published. What do you plan to do during the next reporting period to accomplish the goals?Spring Complete remaining microcosm experiments (Objective 1) Complete data analysis for wetland mesocosms experiments at Western Washington University (Objective 1) WWU MS Graduation (Anticipated completion by March 2024, Objective 2) UK MS graduation (Complete by May 2025, Objectives 2 and 3) Submit peer reviewed journal article 1 (Objective 2) Analyze data collected during the growing season of 2024 and incorporate all data into modeling efforts (Objectives 1, 2, and 3) Summer Submit peer reviewed journal article 2 (Objective 2 and 3) Analyze data collected during the growing season of 2024 and incorporate all data into modeling efforts (Objectives 1, 2, and 3) Fall Analyze data collected during the growing season of 2024 and begin incorporating all data in modeling efforts (Objectives 1, 2, and 3) Winter Submit peer reviewed journal article 3 (Objective 3)

Impacts
What was accomplished under these goals? During the 2023-2024year of the project, completed the field assessment of the pesticide applications for a third year. Further, Washington agricultural and wetland soils were started and are now close to completion by the USDA lab (Lincoln, NE), one Master's graduate student has established the wetland mesocosms at Western Washington University, one Master's graduate student was recruited to the University of Kentucky, all mesocosm experiments were completed at the University of Kentucky. Dr. Tiffany Messer participated in the Gordan Conference Nanoscale Science and Engineering for Agriculture and Food Systems meeting in Summer 2024 and provided an update on the project. Four undergraduates (3 at Western Washington University and 1 at the University of Kentucky) worked on this project and assisted with developing analytical methods to evaluate nanopesticides, establish field plots, and collect and analyze data. Analytical methods for nano-copper and imidacloprid and potential byproducts were finalized and have been used to analyze results for 3 years of the field plots and 2 years of the mesocosm experiments. The following specific activities were completed: Research Objective 1:Complete coupled microcosm/mesocosm scale experiments to assess microbial alterations and neonicotinoid insecticide degradation based on varying nanopesticide application and recurrance rates; 1. Major Activities Completed / Experiments Conducted: Kentucky soil and sediment microcosm incubation experiments are complete at the USDA in Lincoln, NE by co-PI Miller with statistical analysis ongoing. Fresh Washington soil and sediment were collected, and microcosm incubation experiments are ongoing. Established wetland mesocosm experiments were completed at Western Washington University. Analyzed samples from mesocosm experiments. Analyzed data collected during the growing season of 2022, 2023, and 2024 Co-PI Miller conducted a training workshop at the University of Nebraska to train students on protocols for microcosm experiments. 2. Data Collected: Water quality: nano-pesticide, nutrients, bacterial nutrient transformation potential activities Soil: soils samples in mesocosms were analyzed for nutrients, physiochemical parameters, metals, and pesticides Plants: plants samples in mesocosms were analyzed for nutrients, physiochemical parameters, metals, and pesticides 3. Summary Statistics and Discussion of Results: Publication in prep to be submitted Journal of Environmental Quality in Spring/Summer 2025 following the completion of the Washington microcosm experiments focused on implications of nanopesticides to the nitrification and denitrification processes. To date, copper hydroxide was observed to have a notable implication to nitrification. 4. Key Outcomes and Other Accomplishments Realized: 4 conference presentations 1 publicatino in review in Environmental Science: Nano Research Objective 2:Conduct a 2 year field study to assess nanopesticide pesistance, fate, and transport; 1. Major Activities Completed / Experiments Conducted: Analyzed samples from mesocosm and year 3 of field experiments. Sponsored undergraduate research project for student in Summer 2024. MS graduation of Kiley Power in Summer 2024. Analyzed data collected during the growing seasons of 2022 through 2024. 2. Data Collected: Water quality: nano-pesticide, nutrients, bacterial nutrient transformation potential activities Soil: soils samples in field plots were analyzed for nutrients, physiochemical parameters, metals and pesticides Plants: plants samples in field plots were analyzed for nutrients, metals, physiochemical parameters, and pesticides Weather: rainfall duration, volume, and intensity 3. Summary Statistics and Discussion of Results Following the third year of application, plots receiving both bulk and nano-imidacloprid had significant release from field plots following the first rainfall event. No significant differences in release from the fields plots via surface water were observed. Based on Year 1 and 2 evaluations, nanoformulations of copper (II) hydroxide and its interactions with nano-imidacloprid were observed to reduce N and P species export significantly, which supported past findings on impacts to microbial communities that perform nutrient cycling in agroecosystems. However, unlike nutrient observations, the copper (II) hydroxide was not observed to impact runoff concentration, but remained in the soil increasing the mass of soil Cu. These observations emphasize the importance of integrating best management practices to reduce impacts of nutrient losses from agricultural fields receiving copper (II) hydroxide and consideration of prevention mechanisms for Cu buildup in agricultural soil conditions. In the mesocosm experiments, complex interactions between nitrogen, Kocide 3000, and imidacloprid were found to increase nitrate removal rates, decrease phosphate removal rates, and inhibit nitrogen uptake in below-ground biomass. Each treatment, with the exception of the pure control, was found to remove 84 - 99% of nitrate over the sampling period, with removal rates ranging from 0.42 to 1.69 d-1. Imidacloprid was observed to photodegrade but was not completely removed from the wetlands by the end of the sampling period. Large pre-existing copper concentrations in source water led to inconclusive results regarding Kocide 3000 removal. 4. Key Outcomes and Other Accomplishments Realized: 1 completed MS thesis 6 conference presentations 2 publications in prep for submission?to peer reviewed journals Research Objective 3:Create kinetic models from bench, mesocosm, and field scale experiments and incorporate findings into HYDRUS modeling software to predict fate and transport of nanopesticides into the vadose zone 1. Major Activities Completed / Experiments Conducted: MS student at the University of Kentucky has been working on this objective over the past year. 2. Data Collected: • Data to incorporate into the model has been collected for the Kentucky microcosm, mesocosm, and field components of the study and is currently being processed. 3. Summary Statistics and Discussion of Results: Microcosm, mesocosms, and field plots receiving copper (II) hydroxide have been observed to have a significant impact on N degradation and release from soils. 4. Key Outcomes and Other Accomplishments Realized: 1 conference presentation

Publications

  • Type: Theses/Dissertations Status: Published Year Published: 2024 Citation: Kiley Power, (M.S.), Biosystems and Agricultural Engineering, UK. May 2024. Implications of Nanopesticides on Downstream Wetland Ecosystems. August 2022- May 2024. Advisor. Current Position: Engineer at Third Rock (Lexington, KY).
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Stickney, C., Messer, T. L., Montano, M. (Poster) Nanopesticide Fate and Transport in Agroecosystems: A Field Study, Kentucky Water Research Institute Annual Meeting, Lexington, KY, September 27, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Benner, B. Monta�o, M. Evaluating the Efficacy of Soil Amendments in Reducing the Impacts of Nanopesticides, Pacific Northwest Society of Environmental Toxicology and Chemistry, Troutdale, OR, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: 2. Messer, T. L., Montano, M., Miller, D., Rud. W., Powers, K., Stickney, C. Evaluation of Nanopesticide Fate and Transport and Biogeochemical Implications in Agroecosystems, Nanoscale Science and Engineering for Agriculture and Food Systems Gordon Conference, Manchester, NH, June 28, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: 3. Paradis, C.J.; Monta�o, M.D.; Messer, T.; Miller, D. The Effects of nano-Enabled Copper Hydroxide Fungicide on the Fate and Transport of Neonicotinoid Insecticides in a Pacific Northwest Wetland Environment. North American Society of Toxicology and Chemistry, Fort Worth, TX, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Miller, D.N., Messer, T.L., Montano, M. Evaluation of Nanoparticle Impacts on N Cycling in Agricultural Soil and Nearby Wetland Sediment. ASA, CSSA, and SSSA Annual Meeting, San Antonio, TX. November 10, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Power, K.P., Messer, T.L., Rud. W. Implications of Nanopesticides on Downstream Wetland Ecosystems, American Ecological Engineering Society Annual Meeting, Blacksburg, VA, May 31, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Rud, C., Messer, T. L., Montano, M. Impacts of Nanopesticides in Mississippi River Water: Photodegradation Rates, Byproduct Formations, and Degradation Pathways, American Society of Agricultural and Biological Engineers Annual International Meeting, Anaheim, CA, July 30, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Rud, W., Messer, T.L., Montano, M. Field Scale Agricultural Applications of Nanopesticides for Fate, Transport, and Impact to Nutrient Cycling. SouthOn+ Southeast Regional Research Symposium, Lexington, KY. March 25-27, 2024.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2024 Citation: Stickney, C., Messer, T. L., Montano, M. Nanopesticide Fate and Transport in Agroecosystems: A Field Study , American Society of Agricultural and Biological Engineers Annual International Meeting, Anaheim, CA, July 30, 2024.
  • Type: Peer Reviewed Journal Articles Status: Submitted Year Published: 2024 Citation: Goodman, A.J., Benner, B.F.; Monta�o, M.D. Out of the Lab and into the Environment: The Evolution of Single Particle ICP-MS Over the Past Decade, Environmental Science: Nano (in review)


Progress 02/15/23 to 02/14/24

Outputs
Target Audience:During the 2022-2023 year we expanded our outreach programs with Fayette County Public Schools, Perry County Public Schools, Franklin County Public Schools, Hopkins County Public Schools, and the Clover 4-H Extension programs for pre-K and K-12. Updates to the advisory was completed by Dr. Messer to representatives from Kentucky Extension Program and Fayette County Public Schools. Two trainings were completed by Dr. Messer for high school educators who plan to share activities with their classrooms over the next 12 months. Pumpkins grown as part of the project were donated once again to local schools for classroom Halloween projects in addition with sharinginformation regarding the research project. We have presented findings at three conferences (2023 USDA NIFA Annual Meeting (Knoxville, TN), American Ecological Engineering Conference (Florida), American Society of Agricultural and Biological Engineering Conference (Nebraska), and Society of Environmental Toxicology and Chemistry (Kentucky)) and six department seminars (Lexington, KY)).Overall, we reached over 1,000 people throughout outreach events with ages ranging fromundergraduate college students to over 60 years in age.Further, 5 undergraduate and 5 graduate students haveworked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1-3 during the 2022-2023 year and one graduate student completed his MS thesis as part of this project in 2023. Changes/Problems:The USDA lab continued to have a backlog from COVID, which delayed the publication of the microcosm findings from 2021-2022 and the completion of the Washington microcosm experiments. However, co-PD Miller is confident the lab is caught up and we will be able to complete the microcosm experiments this coming year. The publications have been slightly delayed, but we have two completed drafts months away from submission regarding the microcosm Kentucky soils and the Kentucky field plot experiments. What opportunities for training and professional development has the project provided?5 undergraduate and 5 graduate students have worked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1, 2, and 3 in 2022-2023. Students were trained on laboratory analysis, experimental setup, sampling protocol, and all MS students completed their MS proposals related to this project. How have the results been disseminated to communities of interest?Overall, we reached approximately over 1,000 people through outreach events with ages ranging from Pre-K to over 60 years in age.This was completed by presenting findings at four conferences, one webinar, and three department seminars (Lexington, KY)). What do you plan to do during the next reporting period to accomplish the goals?Spring Complete remaining microcosm experiments (Objective 1) Complete wetland mesocosms experiments at Western Washington University (Objective 1) MS graduation (Complete by May 2024, Objective 1) Submit peer reviewed journal article 1 Summer Submit peer reviewed journal article 2 (Objective 2) Analyze samples from Washington mesocosm studies (Objectives 1 and 2) Sponsor undergraduate research project (Objective 1) Recruit PhD/MS student to the University of Kentucky to complete Objective 3 Complete 1 Master's students on the project (2 from the University of Kentucky and 1 from Western Washington University) Fall Analyze data collected during the growing season of 2024 and beging incorporating all data in modeling efforts (Objectives 1, 2, and 3) Winter 2 MS graduation (Complete by December 2023, Objectives 1 and 2) Conduct stakeholders meeting (Complete by December 2024, Objectives 1-3)

Impacts
What was accomplished under these goals? During the 2022-2023 year of the project, completed the field assessment of the pesticide applications for a second year. Further, Washington agricultural and wetland soils were started and are now close to completion by the USDA lab (Lincoln, NE), one Master's graduate student has established the wetland mesocosms at Western Washington University, one Master's graduate student was recruited to the University of Kentucky, all mesocosm experiments were completed at the University of Kentucky. Dr. Tiffany Messer participated in the USDA nano meeting in Summer 2023 and provided an update on the project.Five undergraduates (2 at Western Washington University and 3 at the University of Kentucky) workedon this project and assisted with developing analytical methods to evaluate nanopesticides, establish field plots, and collect and analyze data. Analytical methods for nano-copper and imidacloprid and potential byproducts were finalized and have been used to analyze results for 2 years of the field plots and 1 year of the mesocosms experiments. The following specific activities were completed: * Research Objective 1:Complete coupled microcosm/mesocosm scale experiments to assess microbial alterations and neonicotinoid insecticide degradation based on varying nanopesticide application and recurrance rates; 1. Major Activities Completed / Experiments Conducted: Kentucky soil and sediment microcosm incubation experiments are complete at the USDA in Lincoln, NE by co-PI Miller with statistical analysis ongoing. Fresh Washington soil and sediment were collected, and microcosm incubation experiments are ongoing. Established wetland mesocosms for first growing season at Western Washington University. Analyzed samples from mesocosm experiments. Analyze data collected during the growing season of 2023. Co-PI Miller conducted a training workshop at the University of Nebraska to train students on protocols for microcosm experiments. 2. Data Collected: Water quality: nano-pesticide, nutrients, bacterial nutrient transformation potential activities Soil: soils samples in mesocosms were analyzed for nutrients, physiochemical parameters, metals, and pesticides Plants: plants samples in mesocosms were analyzed for nutrients, physiochemical parameters, metals, and pesticides 3. Summary Statistics and Discussion of Results: Publication in prep to be submitted in a special wetlands collection in the Journal of Natural Resources and Agricultural Systems in Spring/Summer 2024 based on microcosm experiments and implications of nanopesticides to the nitrification and denitrification processes. Copper hydroxide was observed to have a notable implication to nitrification. Publication in prep to be submitted to Journal of Ecological Engineering in Fall/Winter 2024 based on mesocosm experiments and implications of nanopesticides to the nitrification and denitrification processes. Both pesticides were observed to enhance nitrate-N removal as pesticide concentrations increased during the growing season for the Kentucky wetland mesocosms. 4. Key Outcomes and Other Accomplishments Realized: The following outcomes have been identified from this work: Power, K.P.,Messer, T.L., Montaño, M.D. Implications of Nanopesticides on Downstream Wetland Habitats,American Society of Agricultural and Biological Engineers Annual International Meeting, Omaha, NE, July 11, 2023. Messer, T.,Richardson, J., Miller, D. Nanopesticide Influence on Soils,A1511 Project Directors Meeting,Knoxville, TN. Aug 10, 2023. Research Objective 2:Conduct a 2 year field study to assess nanopesticide pesistance, fate, and transport; 1. Major Activities Completed / Experiments Conducted: Analyzed samples from mesocosm and year 2 of field experiments. Sponsored undergraduate research project for student in Summer 2023. MS graduation of William Rud in Summer 2023. Analyze data collected during the growing season of 2023. 2. Data Collected: Water quality: nano-pesticide, nutrients, bacterial nutrient transformation potential activities Soil: soils samples in field plots were analyzed for nutrients, physiochemical parameters, metals and pesticides Plants: plants samples in field plots were analyzed for nutrients, metals, physiochemical parameters, and pesticides Weather: rainfall duration, volume, and intensity 3. Summary Statistics and Discussion of Results Following the first year of application, plots receiving the copper (II) hydroxide had significant increases in release of nitrate-N, total nitrogen and ortho-phosphate regardless of the presence of imidacloprid on the copper (II) hydroxide plots. Further, significant increases in soil copper concentrations in all plots receiving copper (II) hydroxide was also observed. Samples are currently be processed for year 2 of the field plots, with preliminary data indicating potential differences in imidacloprid release and degradation byproducts dependent on plots with varying nanopesticide mixtures. 4. Key Outcomes and Other Accomplishments Realized: William Rud,(M.S.), UK CARES Fellow, Biosystems and Agricultural Engineering, UK. May 2021 - July 2023, Evaluating the Implications of Nanopesticides to N Cycling in Wetland Environments. Current Position: PhD Student at the University of Kentucky (Lexington KY) Rud, W.D.,Messer, T. L., Montaño, M., and Miller, D. (2023) Impact of Nanopesticides in Field Scale Agricultural Applications,American Society of Agricultural and Biological engineers,Omaha, NE, July 11, 2023 Rud, W.D.,Messer, T. L., Montaño, M., and Miller, D. (2023) Nanopesticides in Field Scale Agricultural Applications,American Ecological Engineering Society,Tampa, FL, June 6,2023 Research Objective 3:Create kinetic models from bench, mesocosm, and field scale experiments and incorporate findings into HYDRUS modeling software to predict fate and transport of nanopesticides into the vadose zone; 1. Major Activities Completed / Experiments Conducted: Recruited MS student to the University of Kentucky 2. Data Collected: • Data to incorporate into the model has been collected for the Kentucky microcosm, mesocosm, and field components of the study. 3. Summary Statistics and Discussion of Results: • None to Report 4. Key Outcomes and Other Accomplishments Realized: • None to Report

Publications


    Progress 02/15/22 to 02/14/23

    Outputs
    Target Audience:During the 2021-2022 year we launched continued outreach programs with Fayette County Public Schools, Perry County Public Schools,and the Clover 4-H Extension programs for pre-K and K-12. Updates to the advisory was completed by Dr. Messer to representatives from the pesticide industry (ViveCrop), Kentucky Extension Program, and Fayette County Public Schools. One training was completed by Dr. Messer to over 75 pre-K educators who plan to share with their classrooms over the next 12 months. Pumpkins grown as part of the project were donated to local schools for classroom Halloween projects in addition with sharing information regarding the research project. We have presented findings atthree conferences (USDA NIFA Annual Meeting, American Ecological Engineering Conference, and Society of Environmental Toxicology and Chemistry) and three department seminar (Lexington, KY)).Overall, we reached over 1,000 people throughout outreach events with ages ranging from undergraduate college students to over 60 years in age. Further,5 undergraduate and 3 graduate studentshave worked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1-3 during the 2021-2022 year and one graduated student completed his MS thesis as part of this project. Changes/Problems:Microcosm experiments were slightly delayed due to complication obtaining needed pesticide material to conduct experiments due to COVID19 pandemic. However, experiments are now 50%completed. What opportunities for training and professional development has the project provided?6 undergraduate and 4 graduate students have worked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1 and 2. Students were trained on laboratory analysis, experimental setup, sampling protocol, and all MS students completed their MS proposals related to this project. How have the results been disseminated to communities of interest?Overall, we reached approximately over 1,000 people through outreach events with ages ranging from Pre-K to over 60 years in age.This was completed by presenting findings at four conferences, one webinar, three department seminars (Lexington, KY), and 2 high school presenattions (Washington). What do you plan to do during the next reporting period to accomplish the goals?Spring Complete remaining microcosm experiments (Complete by Summer 2023, Objective 1) Establish wetland mesocosms for first growing season at Western Washington University (Complete by Fall 2023, Objective 1) Complete Year 2 of field experiments and monitoring (Complete by Fall 2023, Objective 2) Summer Submit peer reviewed journal article 1 of this project (Complete by Fall 2023, Objective 1) Analyze samples from mesocosm and year 2 of field experiments (Complete by Winter 2023, Objectives 1 and 2) Sponsor undergraduate research project (Objective 2) MS graduation (Complete by August 2023, Objective 2) Recruit PhD/MS student to the University of Kentucky to complete Objective 3 Fall Analyze data collected during the growing season of 2023 (Complete by December 2023, Objectives 1- 2) Winter Submit peer reviewed journal article 2 of this project (Complete by December 2023, Objective 2) Conduct stakeholders meeting (Complete by December 2023, Objectives 1-3)

    Impacts
    What was accomplished under these goals? During the 2021-2022 year of the project, we installed field sites and completed the field assessment of the pesticide applications. Further, phase 1 of the microcosm experiments for the Kentucky agricultural and wetland soils were completed, one Master's graduate student was recruited to Western Washington University and one Master's graduate student was recruited to the University of Kentucky, and Dr. Tiffany Messer conducted a training and field assessment with Dr. Manuel Montano at the Western Washington University. Dr. Daniel Miller hosted one MS student to be trained on microcosm protocols and preliminary evaluations of the microcosm experiments. Plants for the mesocosm studies were established at the University of Kentucky in fall 2022 and experiments are planned to be initiated in Spring 2023. Dr. Manuel Montano participated in the USDA nano meeting in Summer 2022. Five undergraduates (3at Western Washington University and 3 at the University of Kentucky) worked on this project and assisted with developing analytical methods to evaluate nanopesticides and establish field plots. Analytical methods for nano-copper and imidacloprid and potential byproducts were finalized. The following specific activities were completed: Research Objective 1:Complete coupled microcosm/mesocosm scale experiments to assess microbial alterations and neonicotinoid insecticide degradation based on varying nanopesticide application and recurrance rates; 1. Major Activities Completed / Experiments Conducted: •Kentucky and Washington soil and sediment were sampled and the soil microcosm experiments are ongoing at the USDA in Lincoln, NE by co-PI Miller. Co-PI Montano recruited and trained 3undergraduate research assistants to assist with pesticide detection and quantification methods and 1 Master's graduate student. PI Messer recruited 1 Master's student to conduct the mesocosm scale experiments. Master's graduate Jacob Richardson completed two 1-2 week training trips to the USDA-ARS Lab (Lincoln, NE) during in 2022 and graduate with an MS in Biosystems and Agricultural Engineering in December 2022. PI Messer conducted a training workshop at the Western Washington University to train students and Co-PI Montano on how to conduct mesocosm experiments. Co-PI Miller conducted a training workshop at the University of Nebraska to train students on protocols for microcosm experiments. Co-PI Montano acquired new ICP-MS (NSF-EAR/IF award no. 2138789) to better enable our nano analysis of copper hydroxides. 2. Data Collected: •Water quality: nano-pesticide, nutrients, bacterial nutrient transformation potential activities 3. Summary Statistics and Discussion of Results: •Based on results from this evaluation a publication is in development and planned to be submitted to the Journal of Environmental Pollution in Spring/Summer 2023 based on microcosm experiments and implication of nanopesticides to the nitrification and denitrification processes. Copper hydroxide was observed to have a notable implications to nitrification. 4. Key Outcomes and Other Accomplishments Realized: The following outcomes have been identified from this work: Jake Richardson, (M.S.), NRT Fellow, Biosystems and Agricultural Engineering, UK. Anticipated Completion: December 2022. Nanopesticide Implications on Nitrogen Cycling in Soils. ASABE 2022:Evaluation of Biochar for Sulfate Removal Within Inundated Streams in Appalachia,Jacob Richardson, M.S. Student, Tiffany Messer, PhD Jacob Richardson received the National Science FoundationNational Research Traineeship Award Pratt, K.UK Researcher studying impacts of nanopesticides on Nitrogen. News CAFE UK and features on UKNow, 2021. Messer, T.L.2022. Water Lessons for Preschoolers. Webinar. Co-PI Montano presented at the 2022 Gorden Research Confernce on Nanoscale Science and Engineering for Agriculture and Food Systems Co-PI Montano gave 2 talks at high schools in the region about research completed in this project (Mt. Vernon High School and Sehome High School). Research Objective 2:Conduct a 2 year field study to assess nanopesticide pesistance, fate, and transport; 1. Major Activities Completed / Experiments Conducted: Analyses for the Kentucky field experiments are underway and anticipated to be complete in spring 2023 as part of Master's graduate student William Rud. 2. Data Collected: • Water quality: nano-pesticide, nutrients, metals, pH, specific conductivity, temperature, organic carbon Weather: rainfall Soil Samples: nutrients, metals, pH, physiochemical properties, organic carbon 3. Summary Statistics and Discussion of Results • Samples are currently be processed, with preliminary data indicating potential differences in organic carbon differences between plots with varying nanopesticide treatments 4. Key Outcomes and Other Accomplishments Realized: Rud, K.andMesser, T.L.2022. Fate and Transport of Nanopesticides in Field Scale Agricultural Applications,American Ecological Engineering Society2022 Conference. Baltimore, MD, June 22, 2022.Poster Presentation. Rud, W.D., Messer T. L., Montaño, M.D., Miller, D. N. 2022. Fate and Transport of Nanopesticides in Field Scale Agricultural Applications. Society of Environmental Toxicology and Chemistry. Pittsburgh, Pennsylvania Research Objective 3: Create kinetic models from bench, mesocosm,and field scale experiments and incorporate findings into HYDRUS modeling software to predict fate and transport of nanopesticides into the vadose zone. 1. Major Activities Completed / Experiments Conducted: • None to Report 2. Data Collected: • None to Report 3. Summary Statistics and Discussion of Results: • None to Report 4. Key Outcomes and Other Accomplishments Realized: • None to Report

    Publications

    • Type: Theses/Dissertations Status: Published Year Published: 2022 Citation: Jake Richardson, (M.S.), NRT Fellow, Biosystems and Agricultural Engineering, UK. Anticipated Completion: December 2022. Nanopesticide Implications on Nitrogen Cycling in Soils.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Rud, K. and Messer, T.L. 2022. Fate and Transport of Nanopesticides in Field Scale Agricultural Applications, American Ecological Engineering Society 2022 Conference. Baltimore, MD, June 22, 2022. Poster Presentation.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Rud, W.D., Messer T. L., Monta�o, M.D., Miller, D. N. 2022. Fate and Transport of Nanopesticides in Field Scale Agricultural Applications. Society of Environmental Toxicology and Chemistry. Pittsburgh, Pennsylvania
    • Type: Other Status: Published Year Published: 2022 Citation: Messer, T.L. 2022. Water Lessons for Preschoolers. Webinar.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Messer, T.; Monta�o, M.D.; Miller, D. Evaluation of Nanopesticide Fate and Transport and Biogeochemical Implications in Agroecosystems. Gordon Research Conference: 2022 Nanoscale Science and Engineering for Agriculture and Food Systems, Manchester, NH, 2022.


    Progress 02/15/21 to 02/14/22

    Outputs
    Target Audience:During the 2020-2021 year we launched built collaborations with Fayette County Public Schools, Princeton Public Schools, and the Clover 4-H Extension programs to begin developing materials for outreach for pre-K and K-12 during the 2021-2022, when COVID restrictions are anticipated to become more relaxed. An advisory committee has been formed with representatives from the pesticide industry (ViveCrop), Kentucky Extension Program, Fayette County Public Schools, andWashington Department of Agriculture to assist in evaluating the project's success. We have presented findings at one conference (USDA NIFA Annual Meeting (online) and one department seminar (Lexington, KY)).Overall, we reached approximately 150 people throughout outreach events with ages ranging from undergraduate college students to over 60 years in age. Further, 3 undergraduate and 2 graduate students have worked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1-3 during the 2020-2021 year. Changes/Problems:PI Messer transitioned to a new tenure faculty appointment at the University of Kentucky in October 2020 and the grant transfer was delayed from University of Nebraska - Lincoln to the University of Kentucky until February 2021. Additional delays in the subaward to USDA and Western Washington University resulted in further delaying the microcosm portion of the project until July 2021.Finally, the availability ofone Nanopesticide (nano-imidacloprid) was delayed due to issues arising from material transfer agreements (MTA) between the universities, USDA, and pesticide company. However, the MTA will be finalized in December 2021, with anticipation that the final nanopesticide slated to arrive at the research sites by early 2022 to complete the remaining microcosm studies and use in the upcoming field season. What opportunities for training and professional development has the project provided?3 undergraduate and 2 graduate students have worked on the methodology, analysis, modeling, and peer reviewed publications related to objectives 1 and 2. Students were trained on laboratory analysis, experimental setup, sampling protocol, and MS students Jacob Richardson and William Rud completed their MS proposals related to this project. How have the results been disseminated to communities of interest?Overall, we reached approximately 150 people throughout outreach events with ages ranging from undergraduate college students to over 60 years in age.This was completed by presenting findings at one conference (USDA NIFA Annual Meeting (online) and one department seminar (Lexington, KY)). Further, duringthe 2020-2021 year we launched built collaborations with Fayette County Public Schools, Princeton Public Schools, and the Clover 4-H Extension programs to begin developing materials for outreach for pre-K and K-12 during the 2021-2022, when COVID restrictions are anticipated to become more relaxed. What do you plan to do during the next reporting period to accomplish the goals?Spring Complete remaining microcosm experiments (Complete by Summer 2022, Objective 1) Establish wetland mesocosms for first growing season at University of Kentucky (Complete by Fall 2022, Objective 1) Finalize analytical methods for nano-copper and imidacloprid and potential byproducts (Completed by Summer 2022, Objective 1 and 2) Establish field experiments and begin monitoring (Complete by Fall 2022) Recruit MS student to complete Objective 1-2 of the project to Western Washington University Recruit MS student to complete Objective 3 of the project to the University of Kentucky Summer Submit peer reviewed journal article 1 of this project (Complete by Fall 2022, Objective 1) Analyze samples from microcosm and field experiments (Complete by Winter 2022, Objectives 1 and 2) Sponsor undergraduate research project (Objective 2) Fall Conduct Pre-K through 12 Outreach event (Complete by October 2022) Analyze data collected during the growing season of 2022 (Complete by December 2022, Objectives 1- 2) Winter MS graduation (Complete by December 2022, Objective 1) Conduct stakeholders meeting (Complete by December 2022, Objectives 1-3)

    Impacts
    What was accomplished under these goals? During the 2020-2021 year of the project, we identified and fields sites, established the Kentucky field plots, are conducting phase 1 of the microcosm experiments for the Kentucky agricultural and wetland soils, recruited two Master's graduate students to the University of Kentucky to work on the project, and conducted a workshop with Dr. Manuel Montano at the University of Kentucky. Plants for the mesocosm studies were established at the University of Kentucky in fall 2021. We also participated in the USDA nano meeting virtually in October 2021 and formed a stakeholder committee. Three undergraduates (2 at Western Washington University and 1 at the University of Kentucky) worked on this project and assisted with developing analytical methods to evaluate nanopesticides and establish field plots. Outreach collaborations for Pre-K and K-12 were established, but delayed due COVID19 restrictions at this time. The following specific activities were completed: * Research Objective 1:Complete coupled microcosm/mesocosmscale experiments to assess microbial alterations and neonicotinoid insecticide degradation based on varying nanopesticide application and recurrance rates; 1. Major Activities Completed / Experiments Conducted: •Kentucky soil and sediment were sampled and the Kentucky soil microcosm experiments are ongoing at the USDA in Lincoln, NE by co-PI Miller. Washington soil microcosm experiments are anticipated to begin in spring 2022. Co-PI Montano recruited and trained 2 undergraduate research assistants to assist with pesticide detection and quantification methods. PI Messer recruited 2 Master's students. Students have developed protocol and established field sites for the mesocosm scale experiments. Analyses for the Kentucky soil microcosms are underway and anticipated to be complete in spring 2022. Jacob Richardson completed two 1-2 week training trips to the USDA-ARS Lab (Lincoln, NE) during in 2021. Co-PI Manuel Montano conducted a training workshop at the University of Kentucky to train students and PI Messer on how to assess pesticides using equipment at the University of Kentucky to ensure quality assurance of shipped samples 2. Data Collected: •Water quality: nano-pesticide, nutrients, bacterial nutrient transformation potential activities 3. Summary Statistics and Discussion of Results: •Based on results from this evaluation a publication is in development and planned to be submitted to the Journal of Environmental Pollution in Spring/Summer 2022 with graduate student Jacob Richardson, MS student at the University of Kentucky on this project, as the first author and Dr. Daniel Miller, USDA-ARS, as second author. 4. Key Outcomes and Other Accomplishments Realized: The following outcomes have been identified from this work: Data was presented at national USDA meeting and department seminar Research Objective 2:Conduct a 2 year field study to assess nanopesticide pesistance, fate, and transport; 1. Major Activities Completed / Experiments Conducted: • MS student William Rud, whom is working with PI Messer, has established the field plots and plans to initiate the field study beginning in Spring 2022 2. Data Collected: • None to report 3. Summary Statistics and Discussion of Results • None to report 4. Key Outcomes and Other Accomplishments Realized: • None to report Research Objective 3: Create kinectic models from bench, mesocosm, and field scale experiments and incoporate findings into HYDRUS modeling software to predict fate and transport of nanopesticides into the vadose zone;? 1. Major Activities Completed / Experiments Conducted: • None to Report 2. Data Collected: • None to Report 3. Summary Statistics and Discussion of Results: • None to Report 4. Key Outcomes and Other Accomplishments Realized: • None to Report

    Publications

    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2021 Citation: Messer, T.; Monta�o, M.; Miller, D. Evaluation Of Nanopesticide Fate And Transport And Biogeochemical Implications In Agroecosystem, AFRI Nanotechnology Grantee's Conference, October 6, 2021,
    • Type: Other Status: Accepted Year Published: 2021 Citation: Richardson,J; Messer, T.; Monta�o, M.; Miller, D. Nanopesticide Influence on Soils?, Biosystems and Agricultural Engineering Department Seminar, Lexington, KY, December 3, 2021,