Progress 03/15/11 to 03/14/17

Outputs
Target Audience:The target audience for this project includes agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, local and community gardeners, and citizens concerned about the safety of the food supply. Changes/Problems:During the project, there were no major changes to the research objectives. A scope request change was approved USDA NIFA regarding the inclusion of a vertebrate predator in a trophic transfer experiment that has been completed; see above under objective 3 for additional details. Minor modifications to the technical approach were adopted over the course of the project and were described in the appropriate mid-year or annual reports. Taken together, these modifications were minor and were established to ensure and maximize project success. As mentioned previously, only one significant change in the program occurred. This involved one CoPD switching institutions in year 5; a no-cost extension for the 6th year was granted by USDA to accommodate the effects of this change. We note that the PD and several of the Co-PDs formed a number of new collaborations that allowed us to leverage resources for the work (hence the large number of publications and presentations). In all cases, these collaborations involve separate lines of research that branched off from work in the current project. Specific information can be found in past annual reports. What opportunities for training and professional development has the project provided?A number of Post-doctoral Associates or graduate students were critical to the success of this project (see breakdown below). Each obtained valuable educational and research experience as a result of being funded off of this project. Research findings from the project were presented by project staff (including Graduate Students and Post-doctoral Associates) at a number of educational venues (see lists in previous annual reports). PD White and others have presented this work to several non-technical/public audiences, including public "open house" events in CT, as well as to a number of scientific organizations both nationally and internationally. Several project Post-doctoral Associates and graduate students were awarded NIEHS PhytoScholars Grants from the International Phytotechnology Society (IPS) to attend and present posters at a number of the International Phytotechnologies Conferences, including several international locations. The awards funded travel costs, registration and lodging for the student/post-doc awardees. Project Graduate Students and Post-doctoral Associates CAES- Dr. Roberto De La Torre-Roche (Post-doc).<!-- -->< >- Dr. Yingqing Deng (Graduate student; graduated May 2016).<!-- --> SUNY ESF- Dr. Azam Noori (Post-doc; partial support) Ms. Wenjun Cai (Graduate student; will graduate May 2017) SIU/Texas A &M- Dr. Qiang Wang (Graduate student; graduated December 2014). Mr. Bryan Quan (Graduate student; graduated with MS December 2014). Ms. Yanhui Zheng (Graduate student; graduated with MS December 2014). Dr. Lorenzo Rossi (Post-doc; partial support). How have the results been disseminated to communities of interest?During the six years of our project, significant science impacts were achieved. As highlighted below, 47 peer reviewed publications have been published as part of this project. Four additional publications are currently under review. In addition, since project inception, investigators (or Graduate Students/Post-doctoral Associates) presented project research on 132 occasions at national and international venues. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? General Statement of Performance- This final report summarizes all activities from our 6 years of work (5 years on the grant, one year no-cost extension). In accordance with our previous annual reports for years 1-5, as well as our mid-year report for year 6, all proposed objectives were successfully achieved. Objective 1- The goals of objective 1 were met. Objective one was designed as a high throughput screening study assessing the acute toxicity of 12 NM, as well as the corresponding bulk and ion controls, to 12 crop species. Primary endpoints included biomass, transpiration, and element/particle content in root and shoot tissues as a function of exposure. The original goal of the screening study was to isolate the plant-NM systems of greatest concern so as to guide detailed mechanistic investigations. As noted previously, we only needed to complete approximately two-thirds of the screen prior to having it serve its intended purpose of highlighting plant-NM combinations worthy of more intense investigation. For this objective, the overall findings clearly demonstrate species- and particle type-specific phytotoxicity, as well as concentration-dependent effects. But most importantly, particle-size specific toxicity has been observed consistently, with NM often being more toxic and accumulated to a greater extent than equivalent bulk or ion controls. Results from this objective are currently being compiled for manuscript preparation by the SUNY ESF graduate student. Objective 2- The goals of objective 2 were met Objective two investigated the impact of environmental conditions on NM activity. Work conducted under this objective coincided with work on NM/co-contaminant interactions described below under objective 4. Collectively, the work showed that the conditions of exposure dramatically impact particle behavior (aggregation, solubility), availability, accumulation and toxicity. This is particularly evident when comparing hydroponic to non-hydroponic (MS media, vermiculite, soil) conditions but also, factors such as soil type, solution chemistry/nutrient status, organic matter presence/level, and exposure duration/dose were also important to plant response. Last, properties of the nanomaterials such as functionality or coating were also a major factor influencing accumulation and toxicity. Objective 3- The goals of objective 3 were met Objective three focused on the accumulation, toxicity and transfer of nanomaterials within model terrestrial food chains. The goal of all of these experiments was to track the movement of bulk and nanoparticle (NP) elements/metal oxides through several terrestrial trophic levels, with the overall hypothesis that these materials will behave differently as a function of particle size. The data show that the extent to which these materials transfer from soil to herbivore and subsequent carnivore varies with particle type, soil characteristics, duration of exposure, species used, and life stage of the organisms. In some cases, particle size-dependent trophic transfer was observed; in others, it was not. In one particularly notable study, the weathering of the nanomaterial in soil resulted in a size specific transformation that subsequently increased availability of the NP for accumulation (publication 3, next page). This is clearly a finding of significance and warrants further study. Objective 4- The goals of objective 4 were met Objective 4 focused on the interaction of NM with co-existing contaminants. Given that a large number of other organic and inorganic compounds exist (both intentionally and incidentally) in agricultural systems, we hypothesized that potential NM interactions with these co-contaminants could be important to both analytes. In fact, this hypothesis turned out to be correct. As described in our previous reports, our consistent finding is that NM co-exposure can significantly alter the uptake and translocation of organic co-contaminants. This included both legacy pesticides, as well as select pharmaceutical compounds. In most instances, co-contaminant toxicity and accumulation was decreased by NM co-exposure but in select other instances, the reverse was observed. Clearly additional mechanistic studies focused on understanding the basis for co-contaminant interactions are needed.

Publications

• Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Mukherjee, A.; Hawthorne, J.; White, J.C.; Kelsey, J.W. 2017. Nanoparticle Ag co-exposure reduces the accumulation of weathered persistent pesticides by earthworms. Environ. Tox. Chem. doi:10.1002/etc.3698.
• Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Deng, Y.; Eitzer, B.; White, J.C.; Xing, B. 2017. Impact of multiwall carbon nanotubes on the accumulation and distribution of carbamazepine in collard greens (Brassica oleracea). ES: Nano. 4:149-159.
• Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Servin, A.D.; Pagano, L.; Castillo-Michel, H.; De La Torre-Roche, R.; Hawthorne, J.; Hernandez-Viezcas, J.A.; Loredo, R.; Majumdar, S.; Gardea-Torresdey, J.; Parkash-Dhankher, O.; White, J.C. 2017. Weathering in soil increases nanoparticle CuO bioaccumulation within a terrestrial food chain. Nanotoxicol. 11:98-111.
• Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Yue, L.; Ma, C.; Zhan, X.; White, J.C.; Xing, B. 2017. Molecular mechanisms of maize seedling response to La2O3 NPs exposure: Water uptake, aquaporin gene expression and signal transduction. ES: Nano DOI: 10.1039/c6en00487c.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Zhao, Q.; Ma, C.; White, J.C.; Parkash Dhankher, O.; Zhang, X.; Zhang, S.; Xing, B. 2016. Quantitative evaluation of multi-wall carbon nanotube uptake by Arabidopsis thaliana (L.) in the presence of small polar aromatic organic molecules. Carbon 114:661-670.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Lui, H.; Ma, C.; Chen, G.; White, J.C.; Parkash Dhankher, O.; Xing, B. 2017. Titanium dioxide nanoparticles alleviate tetracycline toxicity to Arabidopsis thaliana. ACS Sustain. Chem. Eng. In press.
• Type: Journal Articles Status: Accepted Year Published: 2017 Citation: Rodrigues, S; Dokoozlian, N; Hendren, C.O.; Karn, B.; Mauter, M.; Sadik, O.; Safarpour, M.; Unrine, J.; Viers, J.; White, J.C.; Wiesner, M.R.; Lowry, G.V. 2017. Nanotechnology for sustainable food production: High value opportunities and scientific challenges. ES: Nano DOI: 10.1039/C6EN00573J.

Progress 03/15/16 to 03/14/17

Outputs
Target Audience:The target audience for this project includes agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, local and community gardeners, and citizens concernedabout the safety of the food supply. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Graduate students at UMass, SUNY-ESF and Texas A&M have been actively engaged in research on this project. Notably, one UMass graduate student was awarded her Ph.D. and has graduated. At CAES and SUNY ESF, two Post-doctoral researchers have completed work on the project and are no longer being funded on the grant. At Texas A&M, a Post-doctoral researcher continues to work on the project. Two CAES research technicians are working part time on this project (salary charged on other grants). Also at CAES, several other Post-doctoral Associates being funded off of parallel food safety projects are able to spend time on research related to the current grant. At UMass, one undergraduate student assisted on the project. At SUNY ESF, two undergraduate students continue to work with the graduate student. Dr. J.C. White continues to serve on Ph.D. committee of two graduate students at the University of Texas-El Paso (UTEP) that are engaged in separate research parallel to that of this project. In addition, J.C. White is serving on the Ph.D. Committee (Co-Promoter) of one graduate student from Hasselt University that is engaged in research related to that of this project. Last, Dr. Luca Pagano received a Post-Doctoral Fellowship from the University of Parma and continues to work in the laboratory of J.C. White on research related to the current project; specifically, Dr. Pagano is investigating the molecular response of agricultural crops to NP exposure. For a three month period in mid-2016, a University of Parma Ph.D. student, Mr. Francesco Pasquali, visited CAES (with funding from the University of Parma) and worked with Dr. Pagano on these experiments. Research findings from the current project have been presented by the investigators,Post-doctoral Associates, and Graduate Studentsat a number of educational venues. PD White and others have presented this work to several non-technical/public audiences, including public "open house" events in CT, as well as to a number of scientific organizations both nationally and internationally. Two CAES Post-doctoral Associates, one UMass graduate student, and one SUNY ESF graduate student all were awarded NIEHS PhytoScholars Grants from the International Phytotechnology Society (IPS) to attend andgive platform presentationsat the 13th International Phytotechnologies Conference in Hangzhou China on September 26-29, 2016. The awards funded travel costs, registration and lodging for the student/post-doc awardees. How have the results been disseminated to communities of interest?To disseminate findings, research results were published in peer-reviewed technical journals and in non-technical publications and venues. During the current period, 16 peer reviewed manuscripts were published; 7 additional manuscripts are under preparation. A total of 23 presentations were given at technical and non-technical meetings by project participants. Non-technical face-to-face interactions occurred at an annual CAES open house event. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period, several research activities are planned. We anticipate completing the data analysis on experiments investigating the physiological and molecular response of zucchini to binary combinations of nanoparticles. We hope to submit a manuscript for publication in Environmental Science: Nano before the end of the calendar year. A Post-doctoral Associate at CAES being supported on separate food safety funding will complete an experiment investigating the impact of quantum dot exposure agricultural crops. This staff member, with the assistance of others at CAES, will also complete a review article on nanomaterial/co-contaminant interactions. At Texas A & M, work will continue on objective 2 and will address nanoparticle effects on agricultural crops at different environmental stress conditions. At Texas A & M, work will begin on Objective 4 and will focus on the effect of co-existing environmental pollutants on nanoparticle uptake and vice versa. The focus will be on understanding mechanisms from the perspectives of plant physiology and root anatomy (e.g. the formation of root barriers in the presence of nanoparticles or/and environmental pollutants). We will also investigate the root exudate facilitated dissolution of metallic nanoparticles in plant rhizosphere and determine whether it is a key mechanism for plant uptake of engineered nanoparticles and co-existing contaminants. A second Post-doctoral Associate at CAES being supported on separate food safety funding will complete manuscript preparation on a study addressing the impact of nanomaterial co-exposure on the accumulation of imidicloprid in zucchini. The SUNY ESF graduate student will complete manuscript preparation on the screening study (objective 1). A thirdCAES Post-doctoral Associate being supported on separate food safety fundswill initiate an experiment investigating the trophic transfer of coated and uncoated Ag nanoparticles (bulk and ion controls) in a terrestrial food chain.

Impacts
What was accomplished under these goals? In accordance with our previous annual reports reports, significant progress continues to be made all objectives. Objective 1- Objective one was designed as a high throughput screening study assessing the acute toxicity of 12 NM, as well as the corresponding bulk and ion controls, to 12 crop species. Primary endpoints include biomass, transpiration, and element/particle content in root and shoot tissues as a function of exposure. The original goal of the screening study was to isolate the plant-NM systems of greatest concern so as to guide detailed mechanistic investigations. As noted previously, although the screen is two-thirds complete, it has served its intended purpose of highlighting plant-NM combinations worthy of more intense investigation. During the current period, no additional screening study experiments were initiated. As noted previously, the overall findings from this objective clearly demonstrate species- and particle type-specific phytotoxicity, as well as concentration-dependent effects. More importantly, particle-size specific toxicity has been observed consistently, with the NM often being more toxic and accumulated to a greater extent than equivalent bulk or ion controls. The SUNY Co-PD and graduate student are currently preparing a manuscript with some of this data. Separate from the screening study, experiments evaluating the role of aquaporins in the uptake of nanoparticle La2O3 by maize were completed and a manuscript was submitted to Environmental Science: Nano. In addition, a review article focused on "omics"-based evaluation of metal oxide nanoparticle exposure in plants was completed and submitted to Environmental Science and Technology. Last, studies evaluating the physiological and molecular response of zucchini to binary mixtures of nanoparticles were initiated. Objective 2- Objective two is investigating the impact of environmental conditions on NM activity. Data analysis of studies investigating the effect of solution chemistry (e.g. the characteristics and concentration of natural organic matter [NOM]) and surface properties of CNTs on their phytotoxicity have concluded at Texas A & M and manuscript preparation is nearly complete. A manuscript comparing the impact of NM exposure on mycorrhizal fungi-plant interactions in soil to that in under hydroponic conditions (done at SUNY) was submitted and accepted at the Journal of Nanoparticle Research. Last, two studies exploring the impact of salt stress on nanoparticle uptake and toxicity were completed; one was recently published in Environmental Pollution and a second is currently being written. Objective 3- Our fourth trophic transfer study was completed and has been submitted for publication in Nanotoxicology. As noted inlast years annual report, this study focused on the uptake and trophic transfer of bulk, ionic and NP CuO that had been in soil for 0 or 60 days prior to planting. In addition, the soil was contaminated with weathered chlordane residues, enabling an assessment of co-contaminant interactions (Objective 4). Lettuce was the plant and the herbivore was crickets; the vertebrate carnivore was the Anolis lizard. A fifth investigation at SUNY ESF has also recently been completed. This study examined the uptake of NP Pt by tomato, and subsequent trophic transfer by tomato hornworms. In addition to total elemental uptake by ICP-MS, both Transmission Electron Microscopy (TEM) and Laser Ablation ICP-MS (LA-ICP-MS) were used to determine the elemental form (NP or not) and localization in the plant and hornworm. Objective 4- Co-contaminant interaction investigations have continued this year and all studies highlight the finding from previous years, which is that exposure to engineered nanomaterials significantly impacts the fate of co-existing contaminants. An investigation at UMass assessing the impact of carboxyl functionalization (COOH-) of MWCNT on the uptake of carbamazepine by collard greens was accepted for publication in Environmental Science: Nano. Another set of experiments at UMass addressing the impact of NP TiO2 and tetracycline co-exposure to Arabidopsis was completed and submitted to ACS Sustainable Chemistry and Engineering. At CAES, a study investigating the impact of exposure to NP CeO2 and Ag on the uptake and translocation of imidicloprid by zucchini was completed and manuscript preparation is underway. In addition, work has begun on a review article summarizing the current literature and knowledge base on nanomaterial/co-contaminant interactions; submission to NanoImpact is predicted in the next three months. Last, although not directly part of this project, a series of experiments evaluating the impact of Ag exposure (NP, bulk, ion; coated and uncoated forms) on the accumulation of weathered chlordane and DDE by earthworms wassubmittedand acceptedin Environmental Toxicology and Chemistry.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ma, X.; Wang, Q.; Rossi, L.; Zhang, W. 2015. Cerium oxide nanoparticles and bulk cerium oxide leading to different physiological and biochemical responses in Brassica rapa. Environ. Sci. Technol. DOI 10.1021/acs.est.5b04111.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Wang, Z.Y., L.N. Xu, J. Zhao, X.K. Wang, J.C. White, and B.S. Xing. 2016. CuO nanoparticle interaction with Arabidopsis thaliana: Toxicity, parent-progeny transfer and gene expression. Environ. Sci. Technol. 10.1021/acs.est.6b01017.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Ma, X.; Quah, B. 2016. Effects of surface charge on the fate and phytotoxicity of gold nanoparticles to Phaseolus Vulgaris. J. Food Chem. Nanotechnol. 2(1):57-65.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Ma, X.; Wang, Q.; Lorenzo, R.; Ebbs, S. D.; White, J. C. 2016. Multigenerational exposure to cerium oxide nanoparticles: physiological and biochemical analysis reveals transmissible changes in rapid cycling Brassica rapa. NanoImpact 1:46-54.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Majumdar, S.; Trujillo-Reyes, J.; Hernandez-Vieczas, J.; White, J.C.; Peralta-Videa, J.; Gardea-Torresdey, J. 2016. Cerium biomagnification in a terrestrial food chain: Influence of particle size and growth stage. Environ. Sci. Technol. 10.1021/acs.est.5b04784.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Mukherjee, A.; Sun, Y.; Morelius, E.; Tamezac, C.; Bandyopadhyay, S.; Niu, G.; White, J.C.; Peralta-Videa, J.R.; Gardea-Torresdey, J. 2016. Differential toxicity of bare and hybrid ZnO nanoparticles in green peas (Pisum sativum L.): A life cycle study Front. Plant Sci. DOI: 10.3389/fpls.2015.01242.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Mukherjee, A.; Majumdar, S.; Servin, A.; Pagano, L.; Parkash-Dhanker, O.; White, J.C. 2016. Carbon nanomaterials in agriculture: A critical review. Front. Plant Sci. http://dx.doi.org/10.3389/fpls.2016.00172.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Servin, A.D.; White, J.C. 2016. Nanotechnology and agriculture: Next steps for understanding the balance between applications and implications. NanoImpact 1:9-12.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Ebbs, S.; Bradfield, S.; Kumar, P.; White, J.C.; Ma, X. 2016. Modeling dietary intake of zinc and copper from consumption of carrot (Daucus carota) exposed to metal oxide nanoparticles and metal ions. Front. Plant. Sci. 3:114-126.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Bradfield, S.; Kumar, P.; White, J.C.; Ebbs, S. 2016. Zinc, copper, or cerium accumulation from metal oxide nanoparticles or ions in sweet potato: Yield effects and projected dietary intake from consumption. Plant Phys. Biochem. doi:10.1016/j.plaphy.2016.04.008.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Dan, Y.; Ma, X.; Zhang, W.; Liu, K.; Chady, S.; Shi, H. 2016. Application of single particle ICP-MS for the determination of plant uptake and accumulation of CeO2 nanoparticles. Analyt. Bioanalyt. Chem. 408:5157-5167.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Rossi, L.; Zhang, W.; Lombardini, L.; Ma, X. 2016. The impact of cerium oxide nanoparticles on salt stress responses of Brassica napa L. Environ. Poll. 219:28-36.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Chen, G.; Ma, C.; Mukherjee, A.; Musante, C.; Zhang, J.; White, J.C.; Parkash Dhankher, O.; Xing, B. 2015. Tannic acid alleviates neodymium oxide nanoparticle toxicity to pumpkin: Physiological and molecular mechanisms. Nanotoxicol. 10:1243-1253.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Ma, C.; Liu, H.; Guo, H.; Musante, C.; Coskun, S.H.; Nelson, B.C.; White, J.C.; Dhanker, O.P.; Xing, B. 2016. Defense mechanisms and nutrient displacement in Arabidopsis thaliana upon exposure to CeO2 and In2O3 nanoparticles. ES: Nano. DOI: 10.1039/C6EN00189K.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Pagano, L.; Servin, A.D.; De La Torre-Roche, R.; Mukherjee, A.; Majumdar, S.; Hawthorne, J.; Marmiroli, M.; Maestri, E.; Marra, R.; Parkash Dhankher, O.; Isch, S.M.; White, J.C.; Marmiroli, N. 2016. Molecular response of crop plants to engineered nanomaterial exposure. Environ. Sci. Technol. 50:71987207.
• Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Noori, A.; White, J.C.; Newman, L.A. 2016. Mycorrhizal fungi influence silver uptake and membrane protein gene expression following Ag-NPs exposure. J. Nano. Res. In press.

Progress 03/15/15 to 03/14/16

Outputs
Target Audience:The target audience for this project includes agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, local and community gardeners, and citizens concerned about the safety of the food supply. Changes/Problems:In anemail dated November 4, 2015; USDA NIFA granted a one year no-cost extension to this project, yielding a project termination date of March 14, 2017. What opportunities for training and professional development has the project provided?Graduate students at UMass, SUNY-ESF and Texas A&M are actively engaged in research on this project. At CAES and SUNY ESF, two Post-doctoral researchers are working on this project. At Texas A&M, a new Post-Doctoral researcher joined the group on July 1, 2015. An undergraduate research assistant (senior) has worked on the project during the fall semester. She will join the graduate program at Texas A&M in the spring, 2016. Two CAES research technicians are working part time on this project (salary charged on other grants). At UMass, three undergraduates (one senior, two freshmen) assisted on the project in the last 12 months. At SUNY ESF, two undergraduate students are working with the graduate student, and two additional undergraduate students are working with the Post-doctoral Associate. During the current period, J.C. White joined the Ph.D. committee of a graduate student at the University of Texas-El Paso (UTEP) that is engaged in research parallel to that of this project. In addition, J.C. White is serving on the Ph.D. Committee (Co-Promoter) of two graduate students from Hasselt University that are engaged in research related to that of this project. Last, Dr. Luca Pagano received a Post-Doctoral Fellowship from the University of Parma and has joined the laboratory of J.C. White to engage in research related to the current project; specifically, Dr. Pagano is investigating the molecular response of agricultural crops to NP exposure. Two UTEP graduate students mentioned in previous reports (Dr. Arnab Mukherjee, Dr. Sanghamitra Majumdar) successfully completed their Ph.D. programs and are now working at CAES as a Post-doctoral Associates on related food safety projects. Work from both Dissertations have been or soon will published. J.C. White continues to serve as a Co-Promoter (Committee member) for two Ph.D. students at Hasselt University in Belgium; both research projects are focused on the impact NM on endophytic and rhizosphere populations of agricultural crops. Research findings from the current project have been presented in a number of educational venues. PD White and others have presented this work to several non-technical/public audiences, including public "open house" events in CT, as well as to a number of scientific organizations both nationally and internationally. Two CAES Post-doctoral Associates, one UMass graduate student, and the Post-doctoral Fellow from the University of Parma all were awarded NIEHS PhytoScholars Grants from the International Phytotechnology Society (IPS) to attend and present posters at the 12th International Phytotechnologies Conference in Manhattan KS on September 27-30, 2015. The awards funded travel costs, registration and lodging for the student/post-doc awardees. How have the results been disseminated to communities of interest?To disseminate findings, research results were published in peer-reviewed technical journals and non-technical publications and venues. During the current period, 12 peer reviewed publications and one book chapter were published; 8 additional publications are in preparation or under review. A total of 25 presentations were given at technical and nontechnical meetings by project participants. Non-technical face-to-face interactions occurred at an annual CAES open house event. What do you plan to do during the next reporting period to accomplish the goals?During the next year, several research activities are planned. As noted above, the writing and publication of several additional manuscripts will be completed. We anticipate completing the data analysis on the latest trophic transfer experiment involving both un-aged and weathered bulk and NP CuO as described above.We hope to begin writing a manuscript in the next 6 months. A Post-doctoral Associate in co-PD Newman's lab will complete work on determining the impact of colonization by mycorrhizal fungi on plant uptake of NM. Two Post-doctoral Associates (Dr. Arnab Mukherjee, Dr. Sanghamitra Majumdar) continued to work in the laboratory of PD White during the current period. Both completed their Ph.D. work in the University of Texas-El Paso Department of Chemistry (PD White served on their Dissertation committees) and are being funded off of separate food safety projects at CAES. Dr. Mukherjee is continuing work to develop a method of NP detection called single particle ICP-MS (spICP-MS); this technique involves the modified use of equipment already present in PD White's laboratory.Dr. Majumdar will continue to develop a LC-MS/MS method for proteomic analysis (similar to work she completed at UTEP). Although these new analytical techniques are being developed for separate projects, they will be applied to the current USDA grant and should significantly advance overall project goals and output. The visiting Post-doctoral Fellow (Dr. Luca Pagano) from the University of Parma in Italy is mostly through his one-year Fellowship; the end date is February of 2015. However, it appears a second year of funding will be available through the University of Parma. Although no funding from the current USDA project will be dedicated to Dr. Pagano, the work he will continue to conduct over the course of the next reporting period will apply directly to the goals of this project (NM effects on crop species). Dr. Roberto De La Torre-Roche (the CAES post-doc funded on this project) will initiate a co-contaminant interactions experiment looking at the impact of NP exposure on the uptake and translocation of neonicotinoid insecticides by certain crop plants.

Impacts
What was accomplished under these goals? In accordance with our previous annual reports, significant progress continues to be made on each objective. Objective 1- Objective one is a high throughput screening study assessing the acute toxicity of 12 NM, as well as the corresponding bulk and ion controls, to 12 crop species. The original goal of the screening study was to isolate the plant-NM systems of greatest concern so as to guide detailed mechanistic investigations. Although the screen is two-thirds complete, it has served its intended purpose of highlighting plant-NM combinations worthy of more intense investigation. During the current reporting period, no additional screening study experiments were initiated. For this objective, the overall findings clearly demonstrate species- and particle type-specific phytotoxicity, as well as concentration-dependent effects. But most importantly, particle-size specific toxicity has been observed consistently, with NM often being more toxic and accumulated to a greater extent than equivalent bulk or ion controls. Current efforts continue to address how to assemble and/or partition the huge data set into one or more peer reviewed publications. The UMass and SUNY graduate students, as well as the CAES Post-doctoral Associate, are currently engaged in this process. Objective 2- Objective two is investigating the impact of environmental conditions on NM activity. Similar to previous reports, data generated under this objective during the current period continues to largely coincide with work on NM/co-contaminant interactions described below under objective 4. At UMass, the manuscript describing the study investigating impact of pristine or carboxyl-functionalized carbon nanotubes (CNTs) on the uptake and translocation carbamazepine in collards (Brassica oleracea) grown under hydroponic and soil-based conditions is currently underway. Separately, the data analysis of studies investigating the effect of solution chemistry (e.g. the characteristics and concentration of natural organic matter [NOM]) and surface properties of CNTs on their phytotoxicity is still underway at Texas A & M. Experiments at SUNY ESF comparing the impact of NM exposure on mycorrhizal fungi-plant interactions in soil to that in under hydroponic conditions have been completed and data analysis is underway. In a new set of experiments completed at UMass, the effect of dissolved organic matter (tannic acid or TA) on NP toxicity to pumpkin was evaluated. These results showed that TA alleviates NP toxicity through alteration of the particle surface charge, thus reducing the contact and uptake of particles by pumpkin. In addition, TA was found to promote antioxidant enzymatic activity by elevating the transcript levels of genes involved in ROS scavenging. This study is currently being written up for submission to Nanotoxicology. Objective 3- Three trophic transfer studies have been completed; one was published previously, a second was published during the current period, and a third is currently under review. A fourth investigation using a vertebrate has just been completed at CAES (see below), and work at SUNY ESF is ongoing. The goal of all of these experiments is to track the movement of bulk and nanoparticle (NP) elements/metal oxides through several terrestrial trophic levels, with the overall hypothesis that these materials will behave differently as a function of particle size. The second study involved soil amendment with 0 or 500 mg/kg NP and bulk La2O3 exposed lettuce, with crickets and darkling beetles as the herbivores and mantids as the predators. Here, particle size did not influence accumulation from soil or transfer through the food chain and we are speculating that element-specific ion dissolution explains this difference from the previous Ce study. The third study (currently under review) was conducted in conjunction with collaborators at the University of Texas-El Paso, using NP and bulk CeO2 with a Texas soil, kidney bean and the bean beetle as the herbivore. A fourth study was recently completed at CAES. The focus was on the uptake and trophic transfer of bulk, ionic and NP CuO that has been in soil for 0 or 60 days prior to planting. In addition, the soil is contaminated with weathered chlordane residues, enabling an assessment of co-contaminant interactions (Objective 4). Lettuce was the plant and the herbivore was crickets; the vertebrate carnivore was the Anolis lizard. All aspects of the experiment have been completed; sample extraction/digestion and analysis is ongoing. Last, ongoing studies at SUNY ESF are examining the uptake of NP Pt by tomato, and subsequent trophic transfer by tomato hornworms. In addition to total elemental uptake by ICP-MS, both Transmission Electron Microscopy (TEM) and Laser Ablation ICP-MS (LA-ICP-MS) are being used to determine the elemental form (NP or not) and localization in the plant and hornworm. Objective 4- Co-contaminant interaction investigations have continued in year five. As described previously, five separate publications have been dedicated to work under this objective (years 2012-2014), with the consistent finding being that NM co-exposure can significantly alter the uptake and translocation of organic co-contaminants. A sixth set of experiments was completed by a UMass graduate student last reporting period and is currently being written up for submission to a peer reviewed journal. The study assessed the impact of carboxyl functionalization (COOH-) of MWCNT on the uptake of carbamazepine by collard greens. In addition to toxicity, carbamazepine and metabolites were quantified in plant tissues by LC/MS-MS. The study found that all CNTs alleviated stress/toxicity in collards from carbamazepine exposure, as well as reducing the in planta accumulation of the chemical by nearly an order of magnitude. In a new set of experiments initiated at UMass, the impact of co-exposure to NP TiO2 and tetracycline on rice and Arabidopsis is being evaluated. Preliminary findings show that NP co-exposure significantly reduces the toxicity of tetracycline; current investigations are focused on evaluated the mechanistic basis for these interactions, including a focus on the expression levels of key detoxication and stress-response genes.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Guo, H.; Zhang, Z.; Xing, B.; Mukherjee, A.; Musante, C.; White, J.C.; He, L. 2015. Analysis of silver nanoparticles in antimicrobial products using Surface Enhanced Raman Spectroscopy (SERS). Environ. Sci. Technol. 49:43174324.
• Type: Book Chapters Status: Published Year Published: 2015 Citation: Ma, X., Chen, M. Advancements in nanoparticle detection and characterization in plant tissues. In Innovations in Nanotechnology. Eds. Chowdhury, A. N., Shapter, J., Imran, A. N. Nova Publisher, 2015.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Collin, B.; Auffan, M.; Johnson, A.; Kaur, I.; Keller, A.; Lazareva, A.; Lead, J.; Ma, X.; Merrifield, R.; Svendsen, C.; White, J.C.; Unrine, J.M. 2015. Environmental release, fate and ecotoxicological effects of manufactured ceria nanomaterials. Environ. Sci.: Nano. 1:533-548.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Servin, A.; Elmer, W.; Mukherjee, A.; De la Torre-Roche, R.; Hamdi, H.; White, J.C.; Bindraban, P.; Dimkpa, C. 2015. A review of the use of engineered nanomaterials to suppress plant disease and enhance crop yield. J. Nano. Res. DOI:10.1007/s11051-015-2907-7.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Zhang, W.; Ebbs, S.D.; Musante, C.; White, J.C.; Gao, C.; Ma, X. 2015. Uptake and Accumulation of bulk and nano-sized cerium oxide particles and ionic cerium by radish (Raphanus sativus L.). J. Agric. Food Chem. 63(2):382-90
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Majumdar, S.; Arigi, E.A.; Choi, H.; Trujillo-Reyes, J.; Margez, J.P.F.; Almeida, I.; White, J.C.; Peralta-Videa, J.R.; Gardea-Torresey, J.L. 2015. Environmental effects of nanoceria on seed production of common bean (Phaseolus vulgaris): A proteomic analysis. Environ. Sci. Technol. 10.1021/acs.est.5b03452.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ebbs, S.; Bradfield, S.; Kumar, P.; White, J.C.; Musante, C.; Ma, X. 2015. Accumulation of zinc, copper, or cerium in carrot (Daucus carota) exposed to metal oxide nanoparticles. Environ. Sci.: Nano. DOI: 10.1039/C5EN00161G.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: De la Torre-Roche, R.; Servin, A.; Hawthorne, J.; Xing, B.; Newman, L.A.; Ma, X.; Chen, G.; White, J.C. 2015. Terrestrial trophic transfer of bulk and nanoparticle La2O3 does not depend on particle size. Environ. Sci. Technol. 49:1186611874.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ma, C.; Chhikara, S.; Minocha, R.; Long, S.; Musante, C.; White, J.C.; Xing, B.; Parkash-Dhanker, O. 2015. Reduced silver nanoparticle phytotoxicity in Crambe abyssinica with enhanced glutathione production by overexpressing bacterial ?-glutamylcysteine synthase. Environ. Sci. Technol. 49:10117-10126.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Sillen, W.; Thijs, S.; Abbamondi; G.R.; Janssen, J.; Weyens, N.; White, J.C.; Vangronsveld, J. 2015. Effects of silver nanoparticles on soil microorganisms and maize biomass are linked in the rhizosphere. Soil Biol. Biochem. 91:14-22.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Ma, C.; White, J.C.; Parkash Dhanker, O.; Xing, B. 2015. Metal-based nanotoxicity and detoxification pathways in higher plants. Environ. Sci. Technol. 49:7109?7122.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Quah, B.; Musante, C.; White, J.C.; Ma, X. 2015. Phytotoxicity, uptake and accumulation of silver with different particle sizes and chemical forms. J. Nano. Res. 17:277.

Progress 03/15/14 to 03/14/15

Outputs
Target Audience: The target audiences for this project include agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, local and community gardeners, and citizens concerned about the safety of the food supply. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Dr. Jason C. White of the Analytical Chemistry Department at the CT Agricultural Experiment Station (CAES) is the primary participant in this project. Dr. Roberto De La Torre-Roche is a Post-doctoral Research Scientist working exclusively on this project at CAES. In addition, Mr. Craig Musante, a Scientist in the Analytical Chemistry Department at CAES, is working part time on this project but is being funded separately. Dr. Baoshan Xing of the University of Massachusetts-Amherst (UMass) Stockbridge School of Agriculture is the first Co-Project Director on this grant. At UMass, Ms. Yingqing (Iris) Deng is the graduate student working on this project; Chuanxin Ma is a second student working part time on the project. The second Co-Project Director is Dr. Lee Newman of the State University of New York College of Environmental Science and Forestry (SUNY ESF) Department of Environmental and Forest Biology, as well as of Brookhaven National Laboratory (BNL). At SUNY ESF, Ms. Wenjun Cai is a graduate student working on this project and Dr. Azam Noori is a post-doc that will begin working on the project in January of 2015. The final Co-Project Director is Dr. Xingmao (Samuel) Ma of the Southern Illinois University-Carbondale (SIU) Department of Civil and Environmental Engineering. At SIU, Dennis Wang is the graduate student working on this project. In addition, PD White continued to serve Ph.D. committee of 2 graduate students from the University of Texas El Paso that are working on projects parallel to the current work. One of the graduate students successfully completed his degree in August 2014 and began working as a Post-Doctoral Associate (Dr. Arnab Mukherjee) in PD White’s lab as of September 2014 on related food safety projects. The second graduate student (Dr. Sanghamitra Majumdar) successfully defended her dissertation in December of 2014 and will be joining the laboratory of PD White in January 2015 to work on related food safety projects. PD White is also serving as a Co-Promoter (Committee member) for a Ph.D. student at Hasselt University in Belgium; the research project is focused on the impact of Ag NP on DDE accumulation and endophyte populations. Two CAES Post-doctoral Associates, including the individual directly funded off of this grant, were sent to Grenoble France to conduct analysis on samples at the European Synchrotron Radiation Facility (ESRF). The samples were from experiments conducted under the current grant, although the funding for the training was covered separately by discretionary funds available to PD White. The CAES Post-doctoral associate and one UMass graduate student working on this project were awarded NIEHS PhytoScholars Grants from the International Phytotechnology Society to attend and present posters at the 11th International Phytotechnologies Conference in Heraklion Crete Greece on September 30-October 3, 2014. How have the results been disseminated to communities of interest? In terms of efforts made to disseminate findings and deliver science to a wider audience in the last year, research results have been published in peer-reviewed technical journals and non-technical publications. During the current reporting period, 7 peer reviewed manuscripts and one book chapter were published. A total of 21 presentations of the results were given at technical and nontechnical meetings by project participants. Non-technical interactions with the public at annual CAES open house events also occurred. PD White also gave a number of interviews to various media outlets. What do you plan to do during the next reporting period to accomplish the goals? During the next year, several research activities are planned. The second and third trophic transfer experiments involving both CeO2 and La2O3 will continue. It is hoped that by March 2015, all data for both experiments will be acquired and the preparation of two separate manuscripts will begin. Additional experiments will be planned based on the findings of these two initial studies. The screening study will continue. Now that we have acquired data for two thirds of the experimental matrix, a discussion of how to break up the information into one or more peer reviewed publications has begun. Additional co-contaminant studies will liklely be planned where the impact of NM co-exposure on the activity and availability of currently used agrichemicals will be assessed. Likely candidates include carbon nanotubes as the NM and atrazine as the pesticide. A range of crops will be investigated. A Post-Doctoral Fellow in co-PD Newman’s lab will continue working on determining the impact of colonization by mycorrhizal fungi on plant uptake of NM. Previously supported on an NSF grant, she is now working under partial funding from the current project. One new Post-doctoral Research Associate (Dr. Arnab Mukherjee) has joined the laboratory of PD White (September 2014). A second post-doc is joining in early January 2015. Both are from the UTEP Department of Chemistry and are being funded off of separate food safety projects. Dr. Mukherjee is currently assisting with work related to the trophic transfer experiments, and in the future, will be re-initiating studies to stand up a new method of nanoparticle detection called single particle ICP-MS (spICP-MS). The second post-doc will be tasked with standing up a LC-MS/MS method for fullerene detection in plant tissues. Although these new analytical techniques are being developed for separate projects, they will be applied to the current USDA grant and should significantly advance overall project goals and output. A Post-Doctoral Scientist (Dr. Luca Pagano) from the University of Parma in Italy has received a one-year Fellowship to study abroad and will likely be joining the laboratory of PD White in February of 2015. The Fellowship is from the University of Parma, although PD White will be providing supplemental funding from discretionary sources. Dr. Pagano has expertise in the molecular effects (genomics, proteomics) of NP exposure on biota. Although no funding from the current USDA project will be dedicated to Dr. Pagano, the work he will conduct over the course of the year will apply directly to the goals of this project (nanomaterial effects on crop species). More specific information will be available for the Year 4 Annual Report in April 2015.

Impacts
What was accomplished under these goals? This progress report covers the period from January 1, 2014 to December 15, 2014. In accordance with our previous reports, significant progress continues to be made on each of our four objectives. Objective 1- Objective one is a high throughput screening study assessing the acute toxicity of 12 NM, as well as the corresponding bulk and ion controls, to 12 crop species. Primary endpoints include biomass, transpiration, and element/particle content in root and shoot tissues as a function of exposure. The original goal of the screening study was to isolate the plant-NM systems of greatest concern so as to guide detailed mechanistic investigations. The screen is nearly two-thirds complete. In fact, the rate of progress on completing objective one has continued to slow this year, largely because it has served its intended purpose of highlighting systems worthy of more intense investigation. During the current reporting period, corn and rice were each evaluated with NP/bulk Al2O3; rye was evaluated with NP/bulk SiO2. The biomass and transpiration data is in hand; digestion and ICP-MS analysis is planned for the coming months. For this objective, the findings to date clearly demonstrate species- and particle type-specific phytotoxicity, as well as concentration-dependent effects. But most importantly, particle-size specific toxicity has been observed repeatedly, with NM being more toxic and accumulated to a greater extent than equivalent bulk or ion controls. Current efforts are addressing how to assemble the acquired data into one or more peer reviewed publications. Objective 2- Objective two is investigating the impact of environmental conditions on NM activity. Similar to previous annual reports, data generated under this objective during the current period continues to largely coincide with work on NM/co-contaminant interactions described below under objective 4. Specifically, the impact of pristine/nonfunctionalized or carboxyl-functionalized carbon nanotubes (CNTs) on the uptake and translocation of the pharmaceutical residue carbamazepine was investigated in collards (Brassica oleracea) grown under hydroponic and soil-based conditions. Further details can be found below. Separately, studies directly investigating the effect of solution chemistry (e.g. the characteristics and concentration of natural organic matter [NOM]) and surface properties of CNTs on their phytotoxicity were also conducted at SIU. Data analysis for these experiments is currently ongoing. Last, in studies recently initiated at SUNY ESF, a Post-Doctoral Research Fellow is investigating the role of mycorrhizal fungal interactions in the uptake of nanoparticles by plants grown in soil. Objective 3- One trophic transfer study has been completed and was published during the current reporting period. Several addtitional trophic experiments are currently underway. The goal of this work is to track the movement of bulk and NP elements through several trophic or feeding levels. The published paper involved the addition of 0 or 1000 mg/Kg CeO2 in NP or bulk form to soil. After 28 day of growth, zucchini roots, stems, leaves and fruit contained significantly more Ce when the exposure was in the NP form. Although the concentrations decreased by more than an order of magnitude at each subsequent trophic level, the particle size specific accumulation carried through to the tissues of both herbivorous crickets and carnivorous spiders. The ongoing experiments involve a similar study that described above with NP and bulk La2O3 exposed lettuce, as well as experiments with collaborators at the University of Texas-El Paso using NP and bulk CeO2 with a different soil, different plant (bean) and alternative herbivore (bean beetle). Separately, studies at SUNY ESF are looking at the uptake of NP Pt by plants, and subsequent accumulation in tomato hornworms. In addition to total uptake, both Transmission Electron Microscopy (TEM) and Laser Ablation ICP-MS (LA-ICP-MS) are being used to determine the form (nanoparticle or not) and localization of the element in the worm (i.e., is it in the gut or moving outside the gut lining). Objective 4- Co-contaminant interaction studies have continued in year four. As described previously, five separate publications have been dedicated to work under this objective (years 2012-2014). The consistent finding has been that NM co-exposure can significantly alter the uptake and translocation of organic co-contaminants. A sixth set of experiments being conducted by the UMass graduate student was completed during the current period. The student investigated the impact of carboxyl functionalization (COOH-) of MWCNT on the uptake of carbamazepine (a common pharmaceutical) by collard greens under hydroponic and soil-grown conditions. In addition to toxicity, carbamazepine and metabolites were quantified in plant tissues by LC/MS-MS. Although this contaminant and plant are not mentioned in the current proposal, carbamazepine has been detected in biosolids and wastewater; therefore, this parallel study still falls under the general topic area of NM-organic contaminant interactions. Specifically, pristine and functionalized CNTs helped alleviate stress/toxicity in collards from carbamazepine exposure, as well as reducing the in planta accumulation of the pharmaceutical by nearly an order of magnitude. This study is currently being written up for publication.

Publications

• Type: Journal Articles Status: Published Year Published: 2014 Citation: Gardea-Torresdey, J.L.; Rico, C.M.; White, J.C. 2014. Trophic transfer, transformation, and impact of engineered nanomaterials in terrestrial environments. Environ. Sci. Technol. 48:25262540.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Bai, Y.; Wu, F.; Petersen, E.J.; White, J.C.; Xing, B. 2014. 100 nanometers: A potentially inappropriate threshold for ecological effects of nanoparticles. Environ. Sci. Technol. 48:3098-3099.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Hamdi, H.; De La Torre-Roche, R.; Hawthorne, J.; White, J.C. 2014. Impact of non-functionalized and amino-functionalized multiwall carbon nanotubes on pesticide uptake by lettuce (Lactuca sativa L.). Nanotox. DOI: 10.3109/17435390.2014.907456.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Petersen, E.J.; Henry, T.B.; Zhao, J.; MacCuspie, R.I.; Kirschling, T.L.; Dobrovolskaia, M.A.; Hackley, V.; Xing, B.; White, J.C. 2014. Identification and avoidance of potential artifacts and misinterpretations in nanomaterial ecotoxicity measurements. Environ. Sci. Technol. 48:4226-4246.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Ma, X.; Gao, C. 2014. Uptake and accumulation of engineered nanomaterials and their phytotoxicity to agricultural crops. In: Emerging Nanotechnologies in Agriculture. Eds. Rai, M., Ribeiro, C., Mattoso, L., Duran, N. Springer, In press.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Deng, Y.; White, J.C.; Xing, B. 2014. Interactions between engineered nanomaterials and agricultural crops: Implications for food safety. J. Zhejiang Univ.- SCIENCE A. 15:552-572.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Zhao, J.; Wang, Z.; White, J.C.; Xing, B. 2014. Graphene in the aquatic environment: adsorption, dispersion, toxicity and transformation. Environ. Sci. Technol. 48:9995?10009.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Hawthorne, J.; De la Torre Roche, R.; Xing, B.; Newman, L.A.; Ma, X.; Majumdar, S.; Gardea-Torresdey, J.; White, J.C. 2014. Particle-size dependent accumulation and trophic transfer of cerium oxide through a terrestrial food chain. Environ. Sci. Technol. 48:13102-13109.

Progress 03/15/13 to 03/14/14

Outputs
Target Audience: The target audiences for this project include agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, local and community gardeners, and citizens concerned about the safety of the food supply. Changes/Problems: As described in the previous report, at CAES Mr. Joseph Hawthorne was funded on of this project temporarily (effective August 2012). Effective March 8, 2013, Mr. Hawthorne's funding was moved off of this project and on to an alternative source. What opportunities for training and professional development has the project provided? Dr. Jason C. White of the Analytical Chemistry Department at the CT Agricultural Experiment Station (CAES) is the primary participant in this project. Dr. Roberto De La Torre-Roche is a Post-doctoral Research Scientist working exclusively on this project at CAES. In addition, Mr. Joseph Hawthorne, a Research Technician in the Analytical Chemistry Department at CAES, and Mr. Craig Musante, a Scientist in the Analytical Chemistry Department at CAES, are working part time on this project. Both Mr. Hawthorne and Mr. Musante arebeing funded separately. From December 2012 to May 2013, Dr. Helmi Hamdi of the University of Carthage (Tunisia) visited the laboratory of J.C. White as a Fulbright scholar (funding through CIES). Dr. Hamdi conducted experiments directly related to the current project; he has since returned to Tunisia but collabortive work is ongoing. Dr. Baoshan Xing of the University of Massachusetts-Amherst (UMass) Department of Plant, Soil, and Insect Sciences is the first Co-Project Director on this grant. At UMass, Ms. Yingqing (Iris) Deng is the graduate student working on this project; Chuanxin Ma is a second student working part time on the project. The second Co-Project Director is Dr. Lee Newman of the State University of New York College of Environmental Science and Forestry (SUNY ESF) Department of Environmental and Forest Biology, as well as of Brookhaven National Laboratory (BNL). At SUNY ESF, Ms. Wenjun Cai is a graduate student working on this project and Ms. Beverly Jose Agtuca is an undergraduate student working part time on this project. The final Co-Project Director is Dr. Xingmao (Samuel) Ma of the Southern Illinois University-Carbondale (SIU) Department of Civil and Environmental Engineering. At SIU, Dennis Wang is the graduate student working on this project. By way of additional training,Dr. White, Dr. De LaTorre-Rocheand Dr. Hamdi (Fulbright Scholar) traveled to the National Institute of Standards and Technology (NIST) in Gaithersburg, MD in January of 2013. In addition to giving a lecture, theDr. Whiteand others received one-day of training on a new technique developed by NIST scientists that uses GC/MS/MS or LC/MS/MS techniques to detect DNA base modifications as a function of nanomaterial exposure. In addition, during the current period, PD White joined the Ph.D. committee of 2 graduate students from the University of Texas El Paso that are working on projects parallet to the current work. Similarly, PD White is also serving as a Co-Promoter (Committee member) for a Ph.D. student at Hasselt University in Belgium; the reserach project is focused on the impact of Ag NP on DDE accumulation and endophyte populations. Two SIU and two UMass graduate students working on this project were awarded NIEHS PhytoScholars Grants from the International Phytotechnology Society to attend and present posters at the 10th International Phytotechnologies Conference in Syracuse NY on October 1-4, 2013. How have the results been disseminated to communities of interest? In terms of efforts made to disseminate findings and deliver science to a wider audience, research results have been published in peer-reviewed technical journals and non-technical publications. During the current reporting period, 8 peer reviewed manuscripts have been published. A total of 17 presentations of the results were given at technical and non-technical meetings by project participants. Non-technical interactions with the public at annual CAES open house events also occurred. PD White also gave a number of interviews to various media outlets. What do you plan to do during the next reporting period to accomplish the goals? During the next year, several research activities are planned. Trophic transfer experiments involving both CeO2 and La2O3 will continue. Protocols are described above. It is hoped that by April 2014, all data for both experiments will be acquired and the preparation of two separate manuscripts will begin. Additional experiments will be planned based on the findings of these two initial studies. The screening study will continue. Now that we have acquired biomass, transpiration and element uptake data for 12 plants exposed to Ag, CuO, CeO2, and ZnO; we plan on writing a manuscript. At this point, it is unclear if that will be completed during the next reporting period. A parallel study with UMass investigators on the impact of Ag (ion, bulk, NP) exposure on transgenic and wild type Crambe abyssinica has been completed. The transgenic varieties overexpress the glutathione anti-oxidative defense genes and exhibited significantly greater growth and Ag accumulation potential than wild type plants. This work, which provides insight on the mechanisms of NM phytotoxicity, is currently being assembled and should be submitted for publication before the next annual report. A parallel study conducted by the SIU Co-PD and PD focused on the toxicity of Ag (bulk, ion, NP) to soybean and wheat has been completed. In addition to toxicity parameters, the particle distribution within plant tissues and cellular organelles was determined by scanning electron microscopy (SEM) and ICP-MS. A draft manuscript is currently in preparation and should be submitted in early 2014. PD White hosted Dr. Tomas Vanek, Head of Laboratory of Plant Biotechnologies at the Institute of Experimental Botany in the Czech Republic, for several days in June 2013. PD White and Dr. Vanek have initiated a collaborative project on the molecular response of plants to NM/NP exposure (Landa, P.; Vankova, R.; Andrlova, J.; Hodek, J.; Marsik, P.; Storchova, H.; White, J.C.; Vanek, T. 2012. Nanoparticle-specific changes in Arabidopsis thaliana gene expression after exposure to ZnO, TiO2, and fullerene soot. J. Hazard. Mat. 241-242:55-62). During this visit, several additional lines of investigation on the molecular response of model plants to engineered nanomaterial exposure were planned and are currently underway. PD White is tentatively scheduled to visit Prague (separate travel funds) to follow up on these studies in May 2014. Two new post-doctoral associates are joining the laboratory of PD White in January 2014. Both are from the UTEP Department of Chemistry and are being funded off of separate food safety projects; one FDA and one USDA. The FDA-funded post-doctoral associate has been tasked with standing up a new method of nanoparticle detection called single particle ICP-MS (spICP-MS). This technique involves the modified use of equipment already present in PD White’s laboratory. The second post-doctoral associate has been tasked with standing up a LC-MS/MS method for fullerene detection in plant tissues. Although these new analytical techniques are being developed for separate projects, they will be applied to the current USDA grant and should significantly advance overall project goals and output.

Impacts
What was accomplished under these goals? This progress report covers the period from January 1, 2013 to December 26, 2013. In accordance with our previous reports, significant progress continues to be made on eachofour fourobjectives. Objective 1- Objective one is a high throughput screening study assessing the acute toxicity of 12 NM, as well as the corresponding bulk and ion controls, to 12 crop species. Primary endpoints include biomass, transpiration, and element/particle content in root and shoot tissues as a function of exposure. The original goal of the screening study was to isolate the plant-NM systems of greatest concern so as to guide detailed mechanistic investigations. The screen is over half way complete. In fact, the rate of progress on completing objective one has slowed somewhat this year, largely because it has already begun to serve its intended purpose of highlighting systems worthy of more intense investigation. Specifically, during the last 6 months, experiments completed under this objective include: toxicity results of four additional species for Al2O3, three species for NH2-MWCNT, two species for MWCNT, and two species for fullerenes. Across the entire screening study to date, nanoparticle (NP) Ag, ZnO, CuO and CeO2 have been studied with all 12 crops. TiO2 and Al2O3 have been evaluated with 9 and 8 plant species, respectively. Because of excessive cost, Au NP and single wall carbon nanotubes (SWCNT) have been replaced with carboxyl-functionalized multiwall carbon nanotubes (COOH-MWCNT) and amino-functionalized MWCNT (NH2-MWCNT). This change, in addition to being an economic necessity, will also enable us to gather information on the impact of functionalization on the toxicity of a given nanomaterial. In all instances, toxicity (biomass, transpiration) data have been collected and analyzed. The elemental content analysis as determined by ICP-MS is proceeding; we have acquired data for over half of the shoots and roots of exposed plants. The additional findings in this period support those of previous report; species- and particle type-specific phytotoxicity, as well as concentration dependent effects are observed. But most importantly, particle-size specific toxicity has been observed repeatedly, with NM being more toxic and accumulated to a greater extent than equivalent bulk or ion controls. Objective 2- Objective two is investigating the impact of environmental conditions on NM activity. During the current reporting period, data generated under this objective coincides with an experiment on NM/co-contaminant interactions described in more detail below under objective 4 below. Specifically, humic acid (natural organic matter) amendment was shown to completely reverse the suppression of pesticide uptake by lettuce induced upon NH2-MWCNT exposure. Interestingly, no such humic acid reversal effect was observed for nonfunctionalized MWCNT. Similarly, an earlier study with three crop species showed that humic acid suppressed pesticide uptake in the presence of fullerenes. The role of NM morphology and chemical characteristics (functionalization) in overall phytotoxicity is currently under investigation. In addition, the recent soil-based experiments described in publication 4 below also fall under this objective as results can be directly compared to earlier studies in vermiculite. Objective 3- Two trophic transfer studies have commenced. The goal of both is to track the movement of bulk and NP metal oxides through several trophic levels. The first experiment involves the addition of 0 or 1000 mg/Kg CeO2 in NP or bulk form to soil. Seven day old zucchini seedlings were subsequently planted and grown for 28 days. At harvest, a portion of the root, stem, leaf, and flower tissues were digested and analyzed by ICP-MS for Ce content. In addition, a portion of the leaf tissue was used as food for crickets during a subsequent 14-day exposure. After two weeks of feeding, most crickets are digested and analyzed by ICP-MS for Ce content. However, a portion of the crickets are used as food for mantids for a one week exposure. Preliminary findings have shown that Ce accumulation in exposed zucchini is significantly greater when the oxide is present in NP form. In addition, crickets fed zucchini leaves containing approximately 0.7 ppm Ce from a bulk particle exposure contain about 0.2 ppm in their tissues, although collected feces also contain 0.2 ppm Ce. Mantids consuming those bulk Ce-exposed crickets only contain 0.003 ppm Ce in their tissues, suggesting that trophic transfer from primary to secondary consumers of bulk CeO2 may be minimal. Experiments focusing on NP CeO2 are currently underway, as well as a second trophic transfer experiment involving NP and bulk La2O3 in soil, with lettuce, crickets and wolf spiders as the receptors. Objective 4- Co-contaminant interaction studies have continued in year three. As described previously, two initial studies were published in 2012 and 2013, and noted that NP co-exposure (fullerenes, Ag) significantly altered the accumulation of pesticides (DDE) by several crops grown in vermiculite. A third study has now been published (December 2013); this most recent study was conducted in soil showed that although MWCNT exposure consistently decreased the availability of weathered chlordane and DDx (DDT and metabolites) as expected. However, fullerene co-exposure effects were far more variable, ranging from modest decreases in pesticide bioaccumulation to no effect to actual increases in contaminant uptake by the plant. An additional study focusing on the impact of MWCNT functionalization and humic acid presence on the uptake of chlordane and DDE by lettuce grown in vermiculite has been completed by Dr. Helmi Hamdi. Dr. Hamdi was a visiting Fulbright Scholar from the University of Carthage (Tunisia) that was awarded a fellowship to work with PD White for five months (December 2012-April 2013). The focus of this study was on edible tissues, thereby providing more relevant data with regard to human exposure. The primary result of significance was differential pesticide accumulation as a result of functionalization; the reductions in pesticide accumulation within edible tissues were significantly greater with the non-functionalized (NF) MWCNT. In addition, although the NH2-functionalization still suppressed pesticide accumulation relative to controls, the presence of humic acid completely reversed that effect; a phenomenon not observed with the NF tubes.The mechanisms of these differential effects are now a subject investigation; a manuscript has been submitted to Environment International. One last set of experiments is being conducted by a UMass graduate student in the laboratory of PD White. The student is investigating the impact of carboxyl functionalization (COOH-) of MWCNT on the uptake of carbamazepine (a common pharmaceutical) by collard greens. In addition to toxicity, carbamazepine and metabolites are being quantified tissues by LC/MS-MS. Although this contaminant and plant are not mentioned in the current proposal, this parallel study still falls under the general topic area of NM-organic contaminant interactions.

Publications

• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Koelmel, J.; Leland, T.; Wang, H.H.; Amarasiriwardena, D.; Xing, B. 2013. Investigation of gold nanoparticles uptake and their tissue level distribution in rice plants by laser ablation-inductively coupled-mass spectrometry. Environ. Poll. 174: 222-228.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: De La Torre-Roche, R.; Hawthorne, Musante, C.; Xing, B.; Newman, L.A.; Ma, X.; White, J.C. 2013. Impact of Ag nanoparticle exposure on p,p-DDE bioaccumulation by Cucurbita pepo (zucchini) and Glycine max (soybean). Environ. Sci. Technol. 47:718725.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Kelsey, J.W.; White, J.C. 2013. Effect of C60 fullerenes on the accumulation of weathered p,p-DDE by plant and worm species under single and multi-species conditions. Environ. Toxicol. Chem. 32:1117-123.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Ma, C.; Chhikara, S.; Xing, B.; Musante, C.; White, J.C.; Parkash Dhankher, O. 2013. Physiological and molecular response of Arabidopsis thaliana to nanoparticle cerium and indium oxide exposure. ACS Sustain. Chem. Eng. 1:768-778.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: De La Torre-Roche, R.; Hawthorne, J.; Deng, Y; Xing, B.; Cai, W.; Newman, L.A.; Wang, Q.; Ma, X.; Hamdi, H.; White, J.C. 2013. Multiwalled carbon nanotubes and C60 fullerenes differentially impact the accumulation of weathered pesticides in four agricultural plants. Environ. Sci. Technol. 47:12539-12547.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Wang, H.; Wu, F.; White, J.C.; Holder, P.A.; Xing, B. 2013. Engineered nanoparticles may induce genotoxicity. Environ. Sci. Technol. 47:13212-13214.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Ma, X.; Uddin, S. 2013. Desorption of 1,3,5-trichlorobenzene from multi-walled carbon nanotubes: Impact of solution chemistry and surface chemistry. Nanomaterials 3:289:302.
• Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Wang, Q.; Ebbs, S.E.; Chen, Y.; Ma, X. 2013. Trans-generational impact of cerium oxide nanoparticles on tomato plants. Metallomics 5:753-759.

Progress 03/15/12 to 03/14/13

Outputs
OUTPUTS: Nanomaterials (NM), particles less than 100 nm, possess unique properties as a function of small size. Over 1300 nanomaterial containing products are now commercially available, including pesticide and fertilizer formulations. The current magnitude of exposure and the unique nature of engineered particulates warrant caution, especially given the regulatory assumption that nanomaterials possess equivalent toxicity and risk to corresponding bulk materials. As a consequence of this assumption, little data have been generated on nanomaterial toxicity to plants. Significant progress has been made on objective 1, which is a high throughput screening study assessing the acute toxicity of 12 NM, as well as the corresponding bulk and ion controls, to 12 crop species. The goal of the screening study is to isolate the plant-NM systems of greatest concern so as to guide detailed mechanistic investigations. The screen is nearly half way complete. To date, nanoparticle silver, zinc oxide, copper oxide and cerium oxide have been studied with all crops. From one to five plants have been tested with aluminum oxide, fullerenes, silicon dioxide or titanium oxide. In all cases, toxicity (biomass, transpiration) data have been collected and analyzed. The elemental content data as determined by ICP-MS are proceeding; we have acquired data for about half of the shoots and roots of exposed plants. The true value of the screening study will be evident only when the study is complete. However, current findings indicate species- and particle type-specific phytotoxicity, as well as concentration dependent effects. Most importantly, particle size-specific toxicity has been observed repeatedly, with nanoparticles being more toxic and accumulated to a greater extent than equivalent bulk or ion controls. For example, silver nanoparticles reduced the biomass and transpiration of soybean, tomato, alfalfa, corn and wheat significantly more than did silver ions or bulk particles. This toxicity correlated well with greater silver tissue content of nanoparticle exposed plants. In addition, significant progress has also been made on objective 4 of the proposal, which is quantifying the altered fate and effects of pesticides as a function of NM co-exposure. In a study published late this year, fullerene co-exposure in vermiculite increased DDE accumulation in zucchini, tomato, and soybean by up to 65%. Significant increases in lipid peroxidation where observed in two of the plants and fullerenes were detected in zucchini shoots. In a follow up study of similar design, nanoparticle silver co-exposure was found to generally suppress DDE uptake by soybean and zucchini. Lipid peroxidation was detected in many of the plants but preliminary evidence suggests that silver exposure closed aquaporin channels and eliminated one pathway by which contaminants cross plant cell membranes. Last, a series of seed germination assays has been initiated by a University of New Haven graduate student. These investigations have just begun but are seeking to determine the effect of surface functionalization of multi-walled carbon nanotubes on soybean and zucchini germination. PARTICIPANTS: Dr. Jason C. White of the Analytical Chemistry Department at the CT Agricultural Experiment Station (CAES) is the primary participant in this project. Dr. Roberto De La Torre-Roche is a Post-doctoral Research Scientist working exclusively on this project at CAES. In addition, Mr. Joseph Hawthorne, a Research Technician in the Analytical Chemistry Department at CAES, and Mr. Craig Musante, a scientist in the Analytical Chemistry Department at CAES, are working part time on this project. Mr. Hawthorne is being funded from this project; Mr. Musante is being funded separately. Dr. Baoshan Xing of the University of Massachusetts-Amherst (UMass) Department of Plant, Soil, and Insect Sciences is the first Co-Project Director on this grant. At UMass, Ms. Yingqing (Iris) Deng is the graduate student working on this project. The second Co-Project Director is Dr. Lee Newman of the State University of New York College of Environmental Science and Forestry (SUNY ESF) Department of Environmental and Forest Biology, as well as of Brookhaven National Laboratory (BNL). At SUNY ESF, Ms. Wenjun Cai is a graduate student working on this project and Ms. Beverly Jose Agtuca is an undergraduate student working part time on this project. The final Co-Project Director is Dr. Xingmao (Samuel) Ma of the Southern Illinois University-Carbondale (SIU) Department of Civil and Environmental Engineering. At SIU, Dennis Wang is the graduate student working on this project. Professor Saion Sinha of the University of New Haven and his graduate student, Soumya Rinda, have joined the project in an unfunded collaboration. TARGET AUDIENCES: The target audiences for this project include agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, and local and community gardeners. In terms of efforts made to disseminate findings and deliver science to a wider audience, research results have been published in peer-reviewed technical journals and non-technical publications. Presentations of the results at technical and non-technical meetings have and will continue to occur (approximately ten during the current reporting period). In addition to post-doctoral and graduate student participation, undergraduate student participation is an important component of this project. PROJECT MODIFICATIONS: Mr. Joseph Hawthorne has been hired as a full time Research Technician to work on this project from August 20, 2012 to March 1, 2013. Effective March 1, 2013, Mr. Hawthorne will be transferred to a separate project.

Impacts
Engineered nanomaterials are clearly being released into agricultural ecosystems, but information on fate, exposure and risk from these materials is lacking. Significant preliminary evidence from our laboratories and others suggests that NM exposure negatively impacts agricultural plants and that this is greater than the bulk material effects. In addition, nanomaterial accumulation by agricultural plants seems likely and represents a significant uncharacterized route of human exposure to these particles. This project is quantitatively characterizing the impact of NMs on common food crops, including a mechanistic assessment of toxicity, accumulation, and trophic transfer, as well as the potential effects on the fate of co-existing insecticides and fungicides. This research described in this project crosses multiple disciplines, including plant physiology, toxicology, environmental and analytical chemistry. Such an effort will provide fundamental knowledge to reduce human exposure to nanomaterials through food chain contamination. Separate but related collaborations with colleagues in China (through Co-PD Xing and PD White with Ocean University of China in Qingdao), the Czech Republic (PD White with the Institute of Experimental Botany in Prague), and Italy (PD White with the University of Parma) will help to advance this research and contribute to overall project outputs and outcomes. The collaboration with China has produced one peer review publication this year and the Czech Republic collaboration has yielded one publication; these manuscripts are separate from those listed as publications this year for the current grant.

Publications

• De La Torre-Roche, R.; Hawthorne, J.; Deng, Y; Xing, B.; Cai, W.; Newman, L.A.; Wang, C.; Ma, X.; White, J.C. 2012. Fullerene-enhanced accumulation of p,p-DDE in agricultural crop species. Environ. Sci. Technol. 46:9315−9323.
• Pan, B.; B. Xing. 2012. Applications and implications of manufactured nanoparticles in soils: a review. Eur. J. Soil Sci., 63:437-456.
• Zhu, Z.J.; Wang, H.H.; Yan, B.; Zheng, H.; Jiang, Y.; Miranda, O.R.; Rotello, V.M.; Xing, B.; Vachet, R.W. 2012. Effect of surface charge on the uptake and distribution of gold nanoparticles in four plant species. Environ. Sci. Technol. 46:12391-12398.
• Wang, Z.Y., Xie, X.Y.; Zhao, J.;. Liu, X.Y.; Feng, W.Q.; White, J.C.; Xing, B. 2012. Xylem- and phloem-based transport of CuO nanoparticles in maize (Zea mays L.). Environ. Sci. Technol. 46:4434-4441.
• Atha, D.H.; Wang, H.H.; Petersen, E.J.; Cleveland, D.; Holbrook, R.D.; Jaruga, P.; Dizdaroglu, M.; Xing, B; Nelson; B.C. 2012. Copper oxide nanoparticle mediated DNA damage in terrestrial plant models. Environ. Sci. Technol. 46:1819-1827.

Progress 03/15/11 to 03/14/12

Outputs
OUTPUTS: Nanomaterials (NM), particles less than 100 nm, possess unique properties as a function of particle size. Over 1300 nanomaterial containing products are now commercially available, including pesticide and fertilizer formulations. The current magnitude of exposure and the unique nature of engineered particulates warrant caution, especially given the regulatory assumption that nanomaterials are equivalent to corresponding bulk materials. Consequently, the impact of nanomaterials on plants has only recently been explored. Significant progress has been made on objective 1, which involves a screening study in which the acute toxicity of 12 NMs, as well as the corresponding bulk and ion controls, is being determined for 12 crops. The screening study will highlight which plant-NM systems are most sensitive under hydroponic, sand and soil conditions so as to guide more detailed mechanistic investigations. The 12 plants were divided among the participating institutions to evaluate the toxicity and accumulation of the NMs at 50 and 500 mg/L under hydroponic conditions. To date, nanoparticle silver has been studied with zucchini, alfalfa, corn, rye, rice, tomato, pea, spinach, wheat and soybean. Alfalfa, rye, soybean, pea, tomato and wheat also have been exposed to nanoparticle copper oxide. Soybean, pea, and tomato also have been evaluated with cerium oxide and zinc oxide. The toxicity of silver was found to be dependent on particle size and plant species. For example, silver nanoparticles reduced the biomass and transpiration of soybean and wheat significantly more than did silver ions or bulk particles. This toxicity correlated well with greater amounts of silver in the tissues of nanoparticle exposed plants. However, silver had little impact on zucchini and rice growth and the elemental content of these plants was not impacted by particle size. For wheat exposed to copper oxide, the differential toxicity as a function of particle size was also dependent on concentration. Nanoparticle copper oxide at 50 mg/L reduced wheat biomass significantly more than corresponding bulk amounts but at 500 mg/L exposure, phytotoxicity was equivalent for both particle sizes. Once the screening study is complete, a more thorough characterization of plant-NM interactions will be possible. Progress has also been made on objective 4 of the proposal, which is quantifying the facilitated uptake of pesticides through NM-chemical interactions. In a preliminary study, zucchini was grown in vermiculite containing 0 or 40mg of fullerenes for 20 days, and during growth, plants were watered with a solution containing 100 ng/ml DDE with 0 or 100mg/L humic acid. The DDE content of zucchini shoots in the absence or presence of fullerenes was 81 and 143 ng/g, respectively (significantly different); humic acid further increased shoot DDE levels to 250 ng/g. Based on these findings, the impact of fullerenes and humic acid on DDE accumulation by five plants (zucchini, pumpkin, pea, tomato, bean) is currently being studied. The DDE content of the roots and shoots will be determined and an analytical procedure is being developed for the analysis of fullerenes in plant shoots. PARTICIPANTS: Dr. Jason C. White of the Analytical Chemistry Department at the CT Agricultural Experiment Station (CAES) is the primary participant in this project. Dr. Roberto De La Torre-Roche is a Post-doctoral Research Scientist working exclusively on this project at CAES. In addition, Mr. Joseph Hawthorne, an undergraduate at the University of New Haven, and Mr. Craig Musante, a scientist in the Analytical Chemistry Department at CAES, are working part time on this project (both being funded separately). Dr. Baoshan Xing of the University of Massachusetts-Amherst (UMass) Department of Plant, Soil, and Insect Sciences is the first Co-Project Director on this grant. At UMass, Ms. Yingqing (Iris) Deng is the graduate student working on this project. The second Co-Project Director is Dr. Lee Newman of the State University of New York College of Environmental Science and Forestry (SUNY ESF) Department of Environmental and Forest Biology, as well as of Brookhaven National Laboratory (BNL). At SUNY ESF, Ms. Wenjun Cai is a graduate student working on this project and Ms. Beverly Jose Agtuca is an undergraduate student working part time on this project. The final Co-Project Director is Dr. Xingmao (Samuel) Ma of the Southern Illinois University-Carbondale (SIU) Department of Civil and Environmental Engineering. At SIU, Dennis Wang is the graduate student working on this project. TARGET AUDIENCES: The target audiences for this project include agricultural scientists and engineers, toxicologists, risk assessment professionals, federal and state regulators, consumer protection and product safety professionals, farmers, and local and community gardeners. In terms of efforts made to disseminate findings and deliver science to a wider audience, research results will be published in peer-reviewed technical journals and non-technical publications. Presentations of the results at technical and non-technical meetings have and will continue to occur. In addition to post-doctoral and graduate student participation, undergraduate student participation is an important component of this project. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Engineered nanomaterials are clearly being released into agricultural ecosystems, but information on fate, exposure and risk from these materials is lacking. Significant preliminary evidence from our laboratories and others suggests that NM exposure negatively impacts agricultural plants and that this is greater than the bulk material effects. In addition, nanomaterial accumulation by agricultural plants seems likely and represents a significant uncharacterized route of human exposure to these particles. This project is quantitatively characterizing the impact of NMs on common food crops, including a mechanistic assessment of toxicity, accumulation, and trophic transfer, as well as the potential effects on the fate of co-existing insecticides and fungicides. This research described in this project crosses multiple disciplines, including plant physiology, toxicology, environmental and analytical chemistry. Such an effort will provide fundamental knowledge to reduce human exposure to nanomaterials through food chain contamination. Separate but related collaborations with colleagues in China (through Co-PD Xing and PD White with Ocean University of China in Qingdao), the Czech Republic (PD White with the Institute of Experimental Botany in Prague), and Italy (PD White with the University of Parma) will help to advance this research and to contribute to overall project outputs and outcomes. The collaboration with China has produced two peer review submissions this year and the Czech Republic collaboration has yielded one submission.

Publications

• No publications reported this period