Progress 04/03/05 to 04/02/10

Outputs
Progress Report Objectives (from AD-416) Characterize long term phytoavailability of trace elements in soils amended with swine manure, poultry litter, biosolids, byproducts and composts. Conduct literature review of possible risks from trace elements that have not been evaluated for manure and biosolids and conduct experimental tests needed to provide more complete risk assessments for trace elements in byproducts or contaminated soils. Develop and demonstrate addition of Fe and Mn oxide rich byproducts to manure, biosolids or compost to increase specific metal adsorption capacity and reduce phyto and bio availability of soil accumulated trace elements and phosphate. Develop improved technology for phytoextraction of soil Cd from contaminated soils requiring remediation. Identify methods for bioremediation of munitions contaminated soils using phytoextraction and rumenal biodegradation. Determine if mycorrhizal protein "Glomalin" or soil humic materials give increased metal binding by long term biosolids amended or manured soils and could reduce potential future phytotoxicity of applied metals. Approach (from AD-416) Long-term swine manure amended soils will be sampled with cooperators and the phytoavailability of soil Zn and Cu to sensitive plants as a function of pH will be evaluated; all test and control soils will be adjusted to several pH levels and lettuce grown to evaluate soil element phytoavailability. In addition, methods to increase the amorphous Fe and Mn oxides in manure and other soil amendments will be evaluated with known chemical forms and Fe and Mn rich byproducts from industry. After addition of different test byproducts and Fe/Mn sources, the solubility of metals and phosphate will be evaluated and the treated manures mixed with control soils to test the effect of the oxide additions on element phytoavailability to lettuce. Besides changes in the chemisorption of soil/manure metals on Fe and Mn oxides, increased binding by soil organic matter may increase. One soil organic matter formed by microbes from organic amendments is Glomalin; this will be measured in long term manure or biosolids amended soils. Additionally, spectroscopic methods may be used to characterize changes in soil organic matter ability to bind metals. In the case of soil Cd phytoremediation, phytoextraction by Cd hyperaccumulator plant Thlaspi caerulescens will be developed. Diverse genetic types will be evaluated for properties needed in commercial Cd phytoextraction and improved cultivars will be produced by normal plant breeding. Agronomic practices required to maximize annual Cd removal will be field tested with CRADA cooperator. For some Cd contaminated soils, application of Zn and limestone may reduce Cd phytoavailability by preventing upregulation of Zn uptake which increases Cd accumulation even at neutral soil pH. Cooperators will meet to design experimental program to efficiently utilize available funds to further develop bioremediation of TNT using phytoextraction by plants followed by mineralization by rumen microbes. Cooperators will select plant species for testing after review of previous research on plant uptake of TNT; they will identify any soil management conditions which favor TNT uptake into plant shoots rather than immobilization by chemical reactions in the roots. Because preliminary test showed that chemical TNT could be biodegraded by rumen microbes, additional rumen studies should examine TNT that has been absorbed by plant roots and incorporated in plant shoots to validate that plant-TNT can also be biodegraded in the rumen. Chemically pure 14C-TNT will be used in these tests so that the balance of TNT can be quantitatively assessed. Partial degradation products will be identified as appropriate to determine effectiveness of this bioremediation technology. Cooperated with US-EPA Superfund in testing methods to revegetate a 50- year barren mine waste from producing asbestos in northern Vermont. About 400 A. of barren mine waste is subject to wind and water erosion. Our previous work showing that organic amendments plus alkaline byproducts could support inexpensive remediation and revegetation of Zn mine waste and Zn-smelter killed ecosystems at several locations. The mine wastes are serpentinite rock with Mg silicate (pH 8.5) and severely deficient in Ca and P for plant growth. Based on experience with growing Ni hyperaccumulator plants on serpentine soils in OR, evaluated remediating the soil fertility confirming the multiple severe plant nutrient deficiencies. Conducted greenhouse pot test of a mixture of composted livestock manure with composted yard debris and cafeteria waste, without and with added gypsum. Tested growth of 9 plant species on the control or fertilized mine waste, or compost and compost+gypsum over the mine waste. All species did very poorly or died on the mine waste alone, and very poorly on the fertilized mine waste. With added compost over the mine waste, all species did well, especially tall fescue, Kentucky bluegrass, perennial ryegrass and alsike clover recommended for revegetation of normal soils in that area of the U.S. Leaching of nutrients from the compost into the mine waste layer allowed extensive grown of plant roots into the otherwise hostile mine waste. Method relies on nutrients which can be leached from a surficial compost layer into the mineral mine waste to correct severe Ca deficiency, plus supplying all essential nutrients by the compost layer. Cooperated with US-EPA and other ARS scientists, Ohio State University and the Electric Power Research Institute in conducting a risk evaluation for beneficial use of fluidized gas desulfurizationm (FGD) gypsum produced after removal of fly ash from the exhaust of coal-fired power plants. Statistically evaluated the composition of representative FGD- gypsum from different sources compared to mined gypsum and US background soils, showing that for most elements the FGD-gypsum had lower levels of contaminants than normal soils. FGD-gypsum contained somewhat more Se and Hg than soils. Evaluated potential transfer and risk to humans, livestock and wildlife from all possible exposure pathways. Collected available field results from testing use of FGD-gypsum in the field to use in the risk evaluation and identified data quality needs of the project. Also identified data from other experiments which clarify the risks from trace elements in the FGD-gypsum. The project will continue for two more years, and limited field data are available for evaluation, so no conclusion has been reached at this date. Accomplishments 01 COOPERATED WITH US-EPA IN DEVELOPMENT OF METHODS TO REVEGETATE A 50-YEAR BARREN MINE WASTE FROM PRODUCING ASBESTOS IN NORTHERN VERMONT. About 40 A. of barren mine waste is subject to wind and water erosion. The serpentinite rock mine wastes are principally Mg silicate and severely deficient in Ca and P for plant growth. Evaluated the soil fertility confirming the multiple severe plant nutrient deficiencies, but no evidence of metal toxicities other than Mg. Conducted greenhouse pot tes of a mixture of composted livestock manure with composted yard debris an cafeteria waste, without and with added gypsum. Tested growth of 9 plant species on the control or fertilized mine waste, or compost and compost+gypsum over the mine waste. All species did very poorly or died on the mine waste alone, and very poorly on the fertilized mine waste. With added compost over the mine waste, all species did well, especially tall fescue, Kentucky bluegrass, perennial ryegrass and alsike clover recommended for revegetation of normal soils in that area of the U.S. Leaching of nutrients from the compost into the mine waste layer allowed extensive growth of plant roots into the otherwise hostile mine waste. Severely infertile long barren mine waste from a Superfund asbestos mine waste was readily revegetated using principles of soil remediation developed in this laboratory. Based on these results, ARS will cooperat with US-EPA and the Vermont Department of Environmental Conservation in installation of a field demonstration of revegetation of the mine wastes using local compost resources during this FY. 02 A mixture of composted food waste and yard debris with steel slag used i a compost filter was tested for infiltration rate and effectiveness of metal removal at different flow rates of Pb and Cu highly contaminated building runoff. The alkaline steel slag neutralized acidity generated reaction of Pb and Cu with the compost at even high flow rates tested. The method was sufficiently successful that APHIS funded a field evaluation of the simple and inexpensive technology at the building wher this runoff is generated as well parking lot runoff at the facility. 03 Ground rubber was shown to be a low grade or slow release Zn fertilizer. The kinetics of Zn release from ground rubber compared to other Zn fertilizers was examined showing that rubber Zn was a highly effective slow-release fertilizer for wheat grown on Zn-deficient soil in both pot and an Iranian field. Waste rubber tires comprise a significant problem in all states; this rubber contains about 1.5% Zn, and ground rubber cou be an excellent slow release inexpensive Zn fertilizer for Zn deficient soils.

Impacts
(N/A)

Publications

• Abou-Shanab, R.I., Van Berkum, P.B., Angle, J.S., Delorme, T.A., Chaney, R. L., Ghozlan, H.A., Ghanem, K., Moawad, H. 2010. Characterization of Ni- resistant bacteria in the rhizosphere of the hyperaccumulator alyssum murale by 16S RRNA gene sequence analysis. World Journal of Microbiology and Biotechnology. 26(1):101-108.
• Chaney, R.L., C.L. Broadhurst and T. Centofanti. 2010. Phytoremediation of Soil Trace Elements (Chapter 17). In: Hooda, P. editor. Trace Elements in Soils. Oxford, UK. Blackwell Publ. p. 311-352.
• Kukier, U., Chaney, R.L., Ryan, J.A., Daniels, W.L., Dowdy, R.H., Granato, T.C. 2010. Phytoavailability of Cadmium in Long-Term Biosolids Amended Soils. Journal of Environmental Quality. 39:519-530.
• Mcnear, D.H., Chaney, R.L., Sparks, D.L. 2010. The Metal Hyperaccumulator Alyssum murale Uses Nitrogen and Oxygen Donor Ligands for Ni Transport and Storage. Phytochemistry. 71:188-200.
• Scheckel, K.G., Chaney, R.L., Basta, N.T. and Ryan, J.A. 2009. Advances in assessing bioavailability of metal(loid)s in contaminated Soils. In: Sparks, D.L. editor. Advances in Agronomy. Volume 104. New York, NY. Academic Press. p 1-52.

Progress 10/01/08 to 09/30/09

Outputs
Progress Report Objectives (from AD-416) Characterize long term phytoavailability of trace elements in soils amended with swine manure, poultry litter, biosolids, byproducts and composts. Conduct literature review of possible risks from trace elements that have not been evaluated for manure and biosolids and conduct experimental tests needed to provide more complete risk assessments for trace elements in byproducts or contaminated soils. Develop and demonstrate addition of Fe and Mn oxide rich byproducts to manure, biosolids or compost to increase specific metal adsorption capacity and reduce phyto and bio availability of soil accumulated trace elements and phosphate. Develop improved technology for phytoextraction of soil Cd from contaminated soils requiring remediation. Identify methods for bioremediation of munitions contaminated soils using phytoextraction and rumenal biodegradation. Determine if mycorrhizal protein "Glomalin" or soil humic materials give increased metal binding by long term biosolids amended or manured soils and could reduce potential future phytotoxicity of applied metals. Approach (from AD-416) Long-term swine manure amended soils will be sampled with cooperators and the phytoavailability of soil Zn and Cu to sensitive plants as a function of pH will be evaluated; all test and control soils will be adjusted to several pH levels and lettuce grown to evaluate soil element phytoavailability. In addition, methods to increase the amorphous Fe and Mn oxides in manure and other soil amendments will be evaluated with known chemical forms and Fe and Mn rich byproducts from industry. After addition of different test byproducts and Fe/Mn sources, the solubility of metals and phosphate will be evaluated and the treated manures mixed with control soils to test the effect of the oxide additions on element phytoavailability to lettuce. Besides changes in the chemisorption of soil/manure metals on Fe and Mn oxides, increased binding by soil organic matter may increase. One soil organic matter formed by microbes from organic amendments is Glomalin; this will be measured in long term manure or biosolids amended soils. Additionally, spectroscopic methods may be used to characterize changes in soil organic matter ability to bind metals. In the case of soil Cd phytoremediation, phytoextraction by Cd hyperaccumulator plant Thlaspi caerulescens will be developed. Diverse genetic types will be evaluated for properties needed in commercial Cd phytoextraction and improved cultivars will be produced by normal plant breeding. Agronomic practices required to maximize annual Cd removal will be field tested with CRADA cooperator. For some Cd contaminated soils, application of Zn and limestone may reduce Cd phytoavailability by preventing upregulation of Zn uptake which increases Cd accumulation even at neutral soil pH. Cooperators will meet to design experimental program to efficiently utilize available funds to further develop bioremediation of TNT using phytoextraction by plants followed by mineralization by rumen microbes. Cooperators will select plant species for testing after review of previous research on plant uptake of TNT; they will identify any soil management conditions which favor TNT uptake into plant shoots rather than immobilization by chemical reactions in the roots. Because preliminary test showed that chemical TNT could be biodegraded by rumen microbes, additional rumen studies should examine TNT that has been absorbed by plant roots and incorporated in plant shoots to validate that plant-TNT can also be biodegraded in the rumen. Chemically pure 14C-TNT will be used in these tests so that the balance of TNT can be quantitatively assessed. Partial degradation products will be identified as appropriate to determine effectiveness of this bioremediation technology. Significant Activities that Support Special Target Populations Trace metals are required to sustain plant growth, however, excess metal concentrations in soil can be toxic and require remediation. Soils may also contain high levels of organic pollutants that require mitigation. Certain plants can be used to extract the organic pollutants from the soil and others that have a high affinity for metals can be used to mitigate soils with high metal concentrations. Agricultural and industrial byproducts and wastes can also be utilized in mitigation strategies or can be a source of nutritive trace metal for plants. The risks and benefits of several waste streams were examined. A risk assessment for beneficial use of spent foundry sand (SFS) was conducted in cooperation with the U.S.-EPA Office of Solid Wastes Industrial Materials Recycling team. Analysis showed that SFS from iron, steel, and aluminum casting operations were low in trace elements, as low or lower than background U.S. soils. Extensive discussion was focused on arsenic in SFS because EPA had estimated that the Soil Screening Level was 0.429 mg As/kg dry soil, which is below the normal range of as in U.S. soils. Ultimately it was agreed that levels of As in materials permitted for recycling and beneficial use need not be lower than background U.S. soils, which allows general use of SFS. Also, dioxins in SFS were analyzed and found to be present at very low levels, lower than background soils, which supports beneficial use of these SFS. The risk assessment and guidance document was submitted for external Peer Review by US-EPA. The risk assessment for SFS should serve as an example for development of risk assessment for other byproducts which may be land- applied. A plant uptake test for soil TNT was completed with perennial ryegrass, tall fescue, and orchard grass and a manuscript is in preparation. The three grass species were grown on an Oregon soils amended with labeled- TNT and were harvested repeatedly during the growing season. TNT in the crop was found to be incorporated (covalently-linked) with the organic matter of the plant and thus comprised no risk to the environment. This strategy is being examined for use by the Department of Defense. Technology Transfer Number of Other Technology Transfer: 1

Impacts
(N/A)

Publications

• Fellet, G., Centofanti, T., Chaney, R.L., Green, C.E. 2009. NiO(s) (Bunsenite) is not Available to Alyssum species. Plant and Soil Journal. 319:219-223.
• Broadhurst, C.L., Tappero, R.V., Maugel, T.K., Erbe, E.F., Sparks, D.L., Chaney, R.L. 2008. Nickel and Manganese Accumulation, Interaction and Localization in Leaves of the Ni Hyperaccumulators Alyssum murale and Alyssum corsicum. Plant and Soil Journal. 314:35-48.
• Chaney, R.L., Li, Y., Chen, K., Angle, J., Baker, A.J. 2008. Effects of Cadmium on Nickel Tolerance and Accumulation in Alyssum species and Cabbage Grown in Nutrient Solution. Plant and Soil Journal. 311:131-140.

Progress 10/01/07 to 09/30/08

Outputs
Progress Report Objectives (from AD-416) Characterize long term phytoavailability of trace elements in soils amended with swine manure, poultry litter, biosolids, byproducts and composts. Conduct literature review of possible risks from trace elements that have not been evaluated for manure and biosolids and conduct experimental tests needed to provide more complete risk assessments for trace elements in byproducts or contaminated soils. Develop and demonstrate addition of Fe and Mn oxide rich byproducts to manure, biosolids or compost to increase specific metal adsorption capacity and reduce phyto and bio availability of soil accumulated trace elements and phosphate. Develop improved technology for phytoextraction of soil Cd from contaminated soils requiring remediation. Identify methods for bioremediation of munitions contaminated soils using phytoextraction and rumenal biodegradation. Determine if mycorrhizal protein "Glomalin" or soil humic materials give increased metal binding by long term biosolids amended or manured soils and could reduce potential future phytotoxicity of applied metals. Approach (from AD-416) Long-term swine manure amended soils will be sampled with cooperators and the phytoavailability of soil Zn and Cu to sensitive plants as a function of pH will be evaluated; all test and control soils will be adjusted to several pH levels and lettuce grown to evaluate soil element phytoavailability. In addition, methods to increase the amorphous Fe and Mn oxides in manure and other soil amendments will be evaluated with known chemical forms and Fe and Mn rich byproducts from industry. After addition of different test byproducts and Fe/Mn sources, the solubility of metals and phosphate will be evaluated and the treated manures mixed with control soils to test the effect of the oxide additions on element phytoavailability to lettuce. Besides changes in the chemisorption of soil/manure metals on Fe and Mn oxides, increased binding by soil organic matter may increase. One soil organic matter formed by microbes from organic amendments is Glomalin; this will be measured in long term manure or biosolids amended soils. Additionally, spectroscopic methods may be used to characterize changes in soil organic matter ability to bind metals. In the case of soil Cd phytoremediation, phytoextraction by Cd hyperaccumulator plant Thlaspi caerulescens will be developed. Diverse genetic types will be evaluated for properties needed in commercial Cd phytoextraction and improved cultivars will be produced by normal plant breeding. Agronomic practices required to maximize annual Cd removal will be field tested with CRADA cooperator. For some Cd contaminated soils, application of Zn and limestone may reduce Cd phytoavailability by preventing upregulation of Zn uptake which increases Cd accumulation even at neutral soil pH. Cooperators will meet to design experimental program to efficiently utilize available funds to further develop bioremediation of TNT using phytoextraction by plants followed by mineralization by rumen microbes. Cooperators will select plant species for testing after review of previous research on plant uptake of TNT; they will identify any soil management conditions which favor TNT uptake into plant shoots rather than immobilization by chemical reactions in the roots. Because preliminary test showed that chemical TNT could be biodegraded by rumen microbes, additional rumen studies should examine TNT that has been absorbed by plant roots and incorporated in plant shoots to validate that plant-TNT can also be biodegraded in the rumen. Chemically pure 14C-TNT will be used in these tests so that the balance of TNT can be quantitatively assessed. Partial degradation products will be identified as appropriate to determine effectiveness of this bioremediation technology. Significant Activities that Support Special Target Populations The beneficial use of foundry sand was investigated by Dungan in EMBUL. At the end of his active research work, research agronomists working in this project undertook to develop a risk assessment for beneficial use of spent foundry sand (SFS). Text about risk assessement for soil trace elements in SFS was prepared, and by means of conference calls, we cooperated with the U.S.-EPA Office of Solid Wastes Industrial Materials Recycling team to conduct and summarize the risk assessment and prepare a guidance document. A multi-pathway risk assessment was completed for SFS. Analysis showed that SFS from iron, steel and aluminum casting operations were low in trace elements, as low or lower than background U.S. soils. Extensive discussion was focused on arsenic in SFS because EPA had estimated that the Soil Screening Level was 0.429 mg As/kg dry soil, which is below the normal range of as in U.S. soils. Ultimately it was agreed that levels of As in materials permitted for recycling and beneficial use need not be lower than background U.S. soils, which allows general use of SFS. Also, dioxins in SFS were analyzed and found to be present at very low levels, lower than background soils, which supports beneficial use of these SFS. The risk assessment for SFS should serve as an example for development of risk assessment for other byproducts which may be land-applied. Research on growing seed increases of highly promising genotypes of Thlaspi caerulescens continued. With proper vernalization, all strains evaluated flowered and set seed when returned to warm long days. Seed increase can be managed during breeding to improve this species for commercial Cd phytoextraction. In testing of a new approach for bioremediation of TNT contaminated soils, the uptake of TNT by grasses and bioremediation of TNT by sheep rumen processes were evaluated under a Specific Cooperative Agreement with Oregon State University. In one test, 14C-TNT was fed to sheep to assess absorption and extraction of TNT, retention of TNT in tissues, and metabolism of TNT in rumen or tissues. A manuscript was prepared on the study which shows that essentially all of the TNT which entered the rumen was reduced and cross-linked with feed ingrediets or metabolites in the sheep, which alleviated the potential adverse effects of TNT in the environment. The TNT plant uptake test was completed with perennial ryegrass, tall fescue and orchardgrass and a manuscript is nearly prepared. Three grass species were grown on an OR soil amended with 14C-TNT, and harvested repeatedly during the growing season. 14C-TNT in the crop was found to be covalently- linked with organic matter of the plant and thus comprised no risk to the environment. The progress made addressed the needs of National Program 206 (Goal 5.2.2: Develop agricultural practices that maintain or enhance soil resources, thus ensuring sustainable food, feed, and fiber production while protecting environmental quality) and in the Program Component II on Nutrient Management, and within the Problem Area 3 "Management Tools for Indexing and Evaluating Nutrient Fate and Transport".

Impacts
(N/A)

Publications

• Smith, D.J., Craig, A.M., Duringer, J.M., Chaney, R.L. 2008. Absorption, tissue distribution, and elimination of residues after 2,4,6- trinitro[14C]toluene administration to sheep. Environmental Science and Technology 42(7):2563-2569.
• Siebielec, G., Chaney, R.L., Kukier, U. 2007. Liming to Remediate Ni Contaminated Soils with Diverse Properties and a Wide Range of Ni Concentration. Plant and Soil. 299:117-130.
• Chaney, R.L., Angle, J., Broadhurst, C., Peters, C.A. 2007. Improved Understanding of Hyperaccumulation Yields Commercial Phytoextraction and Phytomining Technologies. Journal of Environmental Quality. 36:1429-1443.

Progress 10/01/06 to 09/30/07

Outputs
Progress Report Objectives (from AD-416) Characterize long term phytoavailability of trace elements in soils amended with swine manure, poultry litter, biosolids, byproducts and composts. Conduct literature review of possible risks from trace elements that have not been evaluated for manure and biosolids and conduct experimental tests needed to provide more complete risk assessments for trace elements in byproducts or contaminated soils. Develop and demonstrate addition of Fe and Mn oxide rich byproducts to manure, biosolids or compost to increase specific metal adsorption capacity and reduce phyto and bio availability of soil accumulated trace elements and phosphate. Develop improved technology for phytoextraction of soil Cd from contaminated soils requiring remediation. Identify methods for bioremediation of munitions contaminated soils using phytoextraction and rumenal biodegradation. Determine if mycorrhizal protein "Glomalin" or soil humic materials give increased metal binding by long term biosolids amended or manured soils and could reduce potential future phytotoxicity of applied metals. Approach (from AD-416) Long-term swine manure amended soils will be sampled with cooperators and the phytoavailability of soil Zn and Cu to sensitive plants as a function of pH will be evaluated; all test and control soils will be adjusted to several pH levels and lettuce grown to evaluate soil element phytoavailability. In addition, methods to increase the amorphous Fe and Mn oxides in manure and other soil amendments will be evaluated with known chemical forms and Fe and Mn rich byproducts from industry. After addition of different test byproducts and Fe/Mn sources, the solubility of metals and phosphate will be evaluated and the treated manures mixed with control soils to test the effect of the oxide additions on element phytoavailability to lettuce. Besides changes in the chemisorption of soil/manure metals on Fe and Mn oxides, increased binding by soil organic matter may increase. One soil organic matter formed by microbes from organic amendments is Glomalin; this will be measured in long term manure or biosolids amended soils. Additionally, spectroscopic methods may be used to characterize changes in soil organic matter ability to bind metals. In the case of soil Cd phytoremediation, phytoextraction by Cd hyperaccumulator plant Thlaspi caerulescens will be developed. Diverse genetic types will be evaluated for properties needed in commercial Cd phytoextraction and improved cultivars will be produced by normal plant breeding. Agronomic practices required to maximize annual Cd removal will be field tested with CRADA cooperator. For some Cd contaminated soils, application of Zn and limestone may reduce Cd phytoavailability by preventing upregulation of Zn uptake which increases Cd accumulation even at neutral soil pH. Cooperators will meet to design experimental program to efficiently utilize available funds to further develop bioremediation of TNT using phytoextraction by plants followed by mineralization by rumen microbes. Cooperators will select plant species for testing after review of previous research on plant uptake of TNT; they will identify any soil management conditions which favor TNT uptake into plant shoots rather than immobilization by chemical reactions in the roots. Because preliminary test showed that chemical TNT could be biodegraded by rumen microbes, additional rumen studies should examine TNT that has been absorbed by plant roots and incorporated in plant shoots to validate that plant-TNT can also be biodegraded in the rumen. Chemically pure 14C-TNT will be used in these tests so that the balance of TNT can be quantitatively assessed. Partial degradation products will be identified as appropriate to determine effectiveness of this bioremediation technology. Significant Activities that Support Special Target Populations Objective 1: The utility of ground rubber (contains 1.5% Zn) as a byproduct with potential value as a Zn-fertilizer was assessed in a pot experiment using Lockwood soil. The phytoavailability to spinach and lettuce of applied Zn from rubber compared to Zn sulfate was estimated by plant uptake of the added Zn. A second value of the test was the potential of the added Zn to reduce plant uptake of Cd from this Cd mineralized soil derived from marine shale. Because Zn in the rubber must be released from the rubber before it can be absorbed by the plant roots, rubber-applied Zn was only about 5% as effective as Zn sulfate in increasing Zn. Over time, this Zn should become increasingly plant available, and will be a low cost source of Zn fertilizer value. Added Zn sulfate reduced Cd in spinach by about 40% when 200 mg Zn was added per kg of soil. Alternative soil management practices to further reduce Cd uptake by leafy vegetables are being assessed, and the persistence of the effect of added Zn evaluated. Thus ground rubber appears to be a slow release Zn fertilizer with high purity Zn. Field tests are required to learn the cost effectiveness of using recycled ground rubber in place of virgin Zn in fertilizers. Objective 2: In cooperation with the International Water Management Institute in Bangkok, Thailand, and Phytoextraction Associates LLC, a field trial of Thlaspi caerulescens phytoextraction of Cd was conducted on land contaminated with Cd and Zn by mine waste discharge to irrigation waters. The best genetic lines available at this time were used in the test. Results to be obtained in September, 2007. Objective 3: In previous work, lettuce uptake of stable isotope labeled Cd in soils from long term biosolids utilization field experiments and control soils from the same experiments was assessed. In the lettuce test, the slope of Cd uptake was significantly lower for biosolids amended soil than control soils. The effect of several soil factors in limiting uptake of Cd was assessed by regression analysis, and soil organic matter was found to contribute to the persistent effect of biosolids application which reduced Cd uptake compared to unamended soil which also received the stable isotope applications. Testing of the possible role of glomalin in this organic matter effect on Cd phytoavailability remain to be conducted. Accomplishments Lockwood soil contained natural Cd accumulated to undesired levels in leafy vegetables. In a pot experiment, addition of Zn sulfate at high rates reduced spinach Cd by 40%, but added ground rubber was less effective in reducing Cd uptake because the Zn is only slowly released from the rubber to the soil. Ground rubber appears to be an inexpensive slow release source of high purity Zn fertilizer which could replace use of virgin Zn in fertilizers. An experiment was undertaken to evaluate phytoextraction of metals from soil which had received excessive metals applications from biosolids before present regulations were established. In each case, the effect of soil pH on plant growth and metal accumulation was assessed. The two phytoextraction species, Alyssum murale (Ni) and Thlaspi caerulescens (Cd and Zn) as well as the test leafy vegetable (Swiss chard) had substantially reduced yields at lower pH, but grew well at pH 6.5 and above despite the massive cumulative biosolids metals applications. Analyses of the plants is in progress. TNT feeding test showed that nearly all dietary TNT was reduced and became covalently linked to the diet matrix, greatly reducing the potential for TNT to enter ground water and comprise environmental risk. This confirms a rumen-beaker test conducted earlier that the highly reducing and organic matter rich environment of the rumen is capable of transforming TNT and reducing potential risk from TNT is environmental soils. Technology Transfer Number of Invention Disclosures submitted: 1 Number of Patent Applications filed: 1 Number of U.S. Patents granted: 1 Number of Non-Peer Reviewed Presentations and Proceedings: 5

Impacts
(N/A)

Publications

• Mcnear, D.H., Chaney, R.L., Sparks, D.L. 2007. The Effects of Soil Type and Chemical Treatment on Nickel Speciation in Refinery Enriched Soils: A Multi-Technique Investigation. Geochimica et Cosmochimica Acta. 71:2190- 2208.
• Stuczynski, T., Siebielec, G., Daniels, W.L., Mccarty, G.W., Chaney, R.L. 2007. Biological Aspects of Metal Waste Reclamation With Sewage Sludge. Journal of Environmental Quality. 36:1154-1162.
• Wang, A., Chaney, R.L., Angle, J.S., Mcintosh, M.S. 2006. Using hyperaccumulator plants to phytoextract soil Cd. In: Mackova, M., Dowling, D.N., Macek, T., editors. Phytoremediation and Rhizoremediation. Dordrecht, Netherlands: Springer Verlag. p. 103-114.
• Wood, B.W., Chaney, R.L., Crawford, M. 2006. Correcting micronutrient deficiency using metal hyperaccumulators: alyssum biomass as a natural product for nickel deficiency correction. HortScience. 41(5):1231-1234.
• Codling, E.E., Mulchi, C.L., Chaney, R.L. 2007. Grain yield and mineral element composition of maize grown on high phosphorus soils amended with water treatment residual. Journal of Plant Nutrition. 30:225-240.
• Abou-Shanab, R.I., Angle, J.S., Chaney, R.L. 2006. Bacterial Inoculants Affecting Nickel Uptake by Alyssum murale From Low, Moderate and High Ni Soils. Soil Biology and Biochemistry. 38:2882-2889.

Progress 10/01/05 to 09/30/06

Outputs
Progress Report 1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? Soils can become contaminated with Cd, Zn, Pb, As, Ni or other elements from agricultural practices (land application of manures, biosolids, composts, byproducts, P-fertilizers, pesticides), deposition of industrial aerosols (smelter fume contamination) and many other sources. High levels of Cu and Zn in swine and poultry manure may comprise phytotoxicity risk to sensitive crops and cause severe limits on cumulative manure application. Mineralized parent materials also generate metal rich soils. If Cd, Mo, Se, or Co are applied or occur naturally in excess, food-chain or feed-chain risks may result, while if soil Pb or As have accumulated in excess, soil ingestion may comprise risk. Additional elements may require evaluation for industrial byproducts considered for beneficial use on cropland. Previous work has shown that inorganic (Fe, Mn) and organic adsorption surfaces in manure or biosolids amended soils persistently reduce the phytoavailability of soil Cd, Zn and Cu compared to additions of metal salts. Organic matter may also be increased in manure and byproduct amended soils and limit Cu phytoavailability, and specific organic compounds such as glomalin may be increased by applied soil amendments. For manures rich in trace elements, it is important to determine whether applied Zn and Cu are likely to cause phytotoxicity after long-term use on cropland and the organic matter is biodegraded. Bioassay by pot tests of element phytoavailability in soils from long term well maintained field plots is needed to characterize how well adsorbents in the manure amended soils limit trace element phytoavailability. For some soils, trace element contamination may limit use in crop production (Cd, Pb), or may comprise risk to children if land use is changed to residential (Pb, As). More evidence is needed to win acceptance of in situ remediation of soils by incorporation of Fe-rich manures or byproducts. Some soils have become contaminated by Cd, especially rice soils, or sources lacking the 100-fold Zn co-contamination with Cd in mine wastes. Some Salinas Valley, CA, soils are rich in Cd from marine shale parent materials and cause production of high Cd vegetable crops which threaten market acceptance. Our research has clarified how such Cd contaminated soils comprise risk to humans who subsist on rice diets. Phytoextraction with hyperaccumulator plants appears to be the only cost-effective method to reduce such food-chain Cd risk. As with the Ni phytomining technology developed and commercialized in earlier research, improved cultivars of the Cd-Zn-hyperaccumulator Thlaspi caerulescens will have to be bred and practices for maximization of annual Cd phytoextraction demonstrated in order to commercialize our existing research. Additional byproducts may be found to provide benefit to growers but may contain trace elements for which risk characterization remains incomplete. Risk assessments need to be prepared for these additional byproducts, biosolids and manures to support State and Federal guidance and regulations for beneficial land application of these soil amendments. 2. List by year the currently approved milestones (indicators of research progress) FY-2006: Prepare manuscripts from ongoing study of reduced phytoavailability of Cd in Fe-rich biosolids amended soils. Collect long term manured soils; measure Zn & Cu phytoavailability. Conduct risk assessment for other elements in byproducts. FY-2007: Evaluate relative contribution of Fe and Mn oxides and organic matter in limiting metal uptake or toxicity in manured soils. Identify more Fe-rich byproducts which could be added to manures and byproducts, and low cost method to make amorphous. FY-2008: Report relative importance of Fe and Mn oxides vs. organic matter changes on Cd, Zn and Cu availability to plants. FY-2009: Prepare guidance for adding Fe and Mn oxides to manures and byproducts to reduce risks from metals and phosphate. Complete publication of risk assessments for elements in byproducts which have not been well evaluated in manure or biosolids. Develop methods to phytoextract soil Cd. FY-2006: Test germplasm to choose parents for breeding. Validate method for soils where method may be used commercially. FY-2007: Make genetic crosses and test progenies for high yield and Cd phytoextraction. Conduct field trial of improved plants. FY-2008: Extend field demonstrations to rice and vegetable soils requiring Cd remediation. Develop efficient harvest method for short plants. FY-2009: Submit patent applications for improved cultivars. Cooperate in commercial use of plants and methods to manage plants. Evaluate change in soil organic matter in long term biosolids amended soils. FY-2006: Evaluate methods to measure Glomalin in high Fe biosolids amended soils. FY-2007: Conduct analysis of soils from long-term biosolids and manure amended soils for levels of Glomalin and other forms of organic matter. FY-2008: Evaluate role of changes in soil organic matter on phytoavailability and potential food-chain transfer of metals in amended soils. FY-2008: Complete manuscripts on the role of organic matter relative to that of Fe and Mn oxides in limiting phytotoxicity and food-chain risk from metals in land applied manure, biosolids and byproducts. 4a List the single most significant research accomplishment during FY 2006. We identified that biosolids high in Fe had low water solubility of phosphate. Questions arose about whether this phosphate remained plant available or had been converted to forms which will not be useful in agriculture. Although adding Fe and Al may be able to reduce the water solubility of soil P and protect surface waters, if the less soluble P cannot be used by plants, the method would be less valuable. An experiment was conducted using soils from long term biosolids amended soils using wheat. All soils had high soil test phosphate, but low water soluble phosphate. Nutrients other than P needed to grow wheat on the soils were added. In all treatments tested, sorbed phosphate supplied adequate P for full yield of the plants. The results indicate that adding Fe or Al to manures or biosolids to reduce P solubility does not prevent plants and mycorrhizae from accessing the insoluble phosphate. 4b List other significant research accomplishment(s), if any. Thlaspi caerulescens grown on rice soils in Thailand grew well in a greenhouse test. A field test of high Cd accumulating southern France genotypes was initiated. Sought parties which need to remove Cd from contaminated or mineralized soils to obtain funding needed to conduct this research and prepared several proposals to text Cd remediation and phytoextraction on soils with high Cd:Zn ratios in addition to the tests of rice soils with geogenic ratio of Cd and Zn (1:100). 5. Describe the major accomplishments to date and their predicted or actual impact. New project just beginning in FY-2005; earlier accomplishments reported in previous project. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Worked to communicate methods to combine byproducts rich in Fe, Mn and Al to manures and excessively manured soils to groups which need this technology. Gave invited presentation to regional and national meetings and discussed how risk assessment for other elements must be conducted so that any byproduct which may be used to limit P solubility will not cause other long term risks. In particular, the need to obtain balance between Fe and Mn to prevent induced-Mn deficiency, and between Ca and Mg to prevent future Mg deficiency must be achieved in byproduct utilization planning. Because more farmers are finding they will have less ability to apply manures on their own land due to excessive accumulated P, interest in using byproducts to reduce P solubility has increased. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Invited to present Cost-effective Phytoextraction of Cd from Rice Soils using Thlaspi caerulescens at the International Symposium of Phytoremediation and Ecosystem Health, Hangzhou, China, Sept. 10-13, 2005. Invited to present Soil metal bioavailability to plants at the Partners in Environmental Technology Technical Symposium and Workshop, SERDP, Washington, DC, Nov. 28-30, 2005. Invited to present Phytoextraction of soil Cd, Ni and Zn using hyperaccumulator plant to alleviate risks of metal contaminated soils requiring remediation at the International Workshop on Current Developments in Remediation of Contaminated Lands Pulawy, Poland, Oct. 27- 29, 2005.

Impacts
(N/A)

Publications

• Sukkariyah, B.F., Evanylo, G., Zelazny, L., Chaney, R.L. 2005. Cadmium, cu, ni, and zn availability in a biosolids-amended piedmont soil years after application. Journal of Environmental Quality. 34(6):2255-2262.
• Zhang, L., Angle, J.S., Delorme, T.A., Chaney, R.L. 2003. Degradation of alyssum murale biomass in soil. International Journal of Phytoremediation. 7(3):169-176.
• Wang, A., Angle, J.S., Chaney, R.L., Mcintosh, M.S. 2006. Soil ph effects on uptake of cd and zn by thlaspi caerulescens. Plant and Soil. 281(1-2) :325-337.
• Wang, A.S., Angle, J.S., Chaney, R.L., Delorme, T.A. 2006. Changes in soil biological activities under reduced soil ph during thlaspi caerulescens phytoextraction. Soil Biology and Biochemistry. 38(6):1451-1461.
• Chaney, R.L., Angle, J.S., Wang, A., Mcintosh, M.S., Reeves, R.D. 2005. Cost-effective phytoextraction of cd from rice soils using thlaspi caerulescens. [abstract]. Abstract book of the International Symposium of Phytoremediation and Ecosystem Health, Hangzhou, China. September 10-13, 2005. pp. 33-34.
• Chaney, R.L., Angle, J.S., Wang, A., Mcintosh, M.S., Broadhurst, L., Reeves, R.D. 2005. Phytoextraction of soil cd, ni and zn using hyperaccumulator plants to alleviate risks of metal contaminated soils requiring remediation. [abstract]. Proceedings of the International Workshop on Current Developments in Remediation of Contaminated Lands. Pulaway, Poland, October 27-29, 2005. p. 39.
• Mcnear, D., Chaney, R.L., Sparks, D.L. 2005. The plant-soil interface: soil nickel speciation and the mechanisms of nickel hyperaccumulation [abstract]. 2005:171-2.
• Tappero, R.V., Peltier, E., Chaney, R.L., Sparks, D.L. 2005. In situ speciation of cobalt in ni/co hyperaccumulator alyssum murale using bulk and micro-focused x-ray absorption spectroscopy. American Society of Agronomy [abstract]. 2005:206-1.
• Chaney, R.L. 2005. Soil metal bioavailability to plants. Partners in Environmental Technology Technical Symposium and Workshop [abstract]. Proceedings of Partners in Environmental Technology Technical Symposium and Workshop, Washington, DC, No. D-15 (T420).
• Angle, J., Zhang, L., Chaney, R.L. 2006. Degradation of alyssum biomass in soil. International Conference on Serpentine Ecology. Sienna, Italy, May 2006. p. 6.
• Chaney, R.L., Broadhurst, C., Mcintosh, M.S., Reeves, R.D., Angle, J. 2006. Phytoextraction of heavy metals with hyperaccumulator plants [abstract]. International Bioavailability Workshop, Seville, Spain. September 11-14, 2006. pp. 65-66.
• Mcnear, D.H., Chaney, R.L., Sparks, D.L. 2006. The influence of soil ni speciation on the phytoremediation potential of soils surrounding an historic Ni refinery at Port Colborne, Ontario, Canada [abstract]. World Congress of Soil Science. Abstract 17158.
• Tappero, R., Chaney, R.L., Sparks, D.L. 2006. Spectromicroscopic investigation of co speciation in a ni/co hyperaccumulator plant used for phytoremediation and phytomining [abstract]. World Congress of Soil Science. Abstract 12480.
• Sukkariyah, B.F., Evanylo, G., Zelazny, L., Chaney, R.L. 2005. Recovery and distribution of biosolids-derived trace metals in a davidson clay loam soil. Journal of Environmental Quality. 34(5):1843-1850.

Progress 10/01/04 to 09/30/05

Outputs
1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Soils can become contaminated with Cd, Zn, Pb, As, Ni or other elements from agricultural practices (land application of manures, biosolids, composts, byproducts, P-fertilizers, pesticides), deposition of industrial aerosols (smelter fume contamination) and many other sources. High levels of Cu and Zn in swine and poultry manure may comprise phytotoxicity risk to sensitive crops and cause severe limits on cumulative manure application. Mineralized parent materials also generate metal rich soils. If Cd, Mo, Se, or Co are applied or occur naturally in excess, food-chain or feed-chain risks may result, while if soil Pb or As have accumulated in excess, soil ingestion may comprise risk. Additional elements may require evaluation for industrial byproducts considered for beneficial use on cropland. Previous work has shown that inorganic (Fe, Mn) and organic adsorption surfaces in manure or biosolids amended soils persistently reduce the phytoavailability of soil Cd, Zn and Cu compared to additions of metal salts. Organic matter may also be increased in manure and byproduct amended soils and limit Cu phytoavailability, and specific organic compounds such as glomalin may be increased by applied soil amendments. For manures rich in trace elements, it is important to determine whether applied Zn and Cu are likely to cause phytotoxicity after long-term use on cropland and the organic matter is biodegraded. Bioassay by pot tests of element phytoavailability in soils from long term well maintained field plots is needed to characterize how well adsorbents in the manure amended soils limit trace element phytoavailability. For some soils, trace element contamination may limit use in crop production (Cd, Pb), or may comprise risk to children if land use is changed to residential (Pb, As). More evidence is needed to win acceptance of in situ remediation of soils by incorporation of Fe-rich manures or byproducts. Some soils have become contaminated by Cd, especially rice soils, or sources lacking the 100-fold Zn co-contamination with Cd in mine wastes. Some Salinas Valley, CA, soils are rich in Cd from marine shale parent materials and cause production of high Cd vegetable crops which threaten market acceptance. Our research has clarified how such Cd contaminated soils comprise risk to humans who subsist on rice diets. Phytoextraction with hyperaccumulator plants appears to be the only cost-effective method to reduce such food-chain Cd risk. As with the Ni phytomining technology developed and commercialized in earlier research, improved cultivars of the Cd-Zn-hyperaccumulator Thlaspi caerulescens will have to be bred and practices for maximization of annual Cd phytoextraction demonstrated in order to commercialize our existing research. Additional byproducts may be found to provide benefit to growers but may contain trace elements for which risk characterization remains incomplete. Risk assessments need to be prepared for these additional byproducts, biosolids and manures to support State and Federal guidance and regulations for beneficial land application of these soil amendments. 2. List the milestones (indicators of progress) from your Project Plan. Objective 1: Characterize and reduce long-term phytoavailability of metals applied in manures and byproducts. At 15 months: Prepare manuscripts from ongoing study of reduced phytoavailability of Cd in Fe-rich biosolids amended soils. Collect long term manured soils; measure Zn & Cu phytoavailability. Evaluate methods to measure Glomalin in high Fe biosolids amended soils. Conduct risk assessment for other elements in byproducts. At 30 months: Evaluate relative contribution of Fe and Mn oxides and organic matter in limiting metal uptake or toxicity in manured soils. Identify more Fe-rich byproducts which could be added to manures and byproducts, and low cost method to make amorphous. By 45 months: Report relative importance of Fe and Mn oxides vs. organic matter changes on Cd, Zn and Cu availability to plants. At 60 months: Prepare guidance for adding Fe and Mn oxides to manures and byproducts to reduce risks from metals and phosphate. Complete publication of risk assessments for elements in byproducts which have not been well evaluated in manure or biosolids. Objective 2: Develop methods to phytoextract soil Cd. By 15 months: Test germplasm to choose parents for breeding. Validate method for soils where method may be used commercially. By 30 months: Make genetic crosses and test progenies for high yield and Cd phytoextraction. Conduct field trial of improved plants. By 45 months: Extend field demonstrations to rice and vegetable soils requiring Cd remediation. Develop efficient harvest method for short plants. By 60 months: Submit patent applications for improved cultivars. Cooperate in commercial use of plants and methods to manage plants. 3b List the milestones that you expect to address over the next 3 years (FY 2006, 2007, and 2008). What do you expect to accomplish, year by year, over the next 3 years under each milestone? FY 2006: Complete preparation of manuscripts on study of reduced phytoavailability of Cd in Fe-rich biosolids amended soils, and the labile pool of Cd in these soils. Both lettuce and wheat studies are completed and being written. With cooperator Evanylo, design experimental studies on long- term manures soils and collect soils from their long-term field plots. With assistance from an investigator from SASL, test methods to recover glomalin from biosolids amended soils and apply to long-term field soils used in earlier experiments. Prepare manuscripts from risk assessment of other elements in byproducts being considered for use in agriculture. FY 2007: Complete collection of Fe and Mn rich byproducts which might be used to reduce P solubility and increase metal adsorption capacity of manured soils. Evaluate relative contribution of Fe and Mn oxides and organic matter in limiting metal uptake or toxicity in manured soils. Prepare manuscript from cooperative study with Davis and Kim on using anaerobic digestion to convert hematite wastes into higher surface area Fe oxides which are more effective in sorbing metals and phosphate. FY 2008: A detailed plan to test use of Cd phytoextraction for phosphate mine wastes soils with high Cd and high Cd:Zn. Such soils allow higher uptake of Cd into plants and regulatory agencies are concerned that this high soil Cd could cause harm to wildlife. Previous experience indicated that appropriate genetic evaluation of collected Thlaspi genotypes needed to be done in the field rather than greenhouse or growth chamber, in order to see the full expression of the potential growth of the plants and their ability to remove soil Cd. Greenhouse or growth chambers could be used to compare additional germplasm as part of the overall testing. In addition, the application of the selected genotypes which do well on the phosphate mine waste will be evaluated on other high Cd soils; such as the high Cd Lockwood soils rich in geogenic Cd and high Cd:Zn ratio, or soils which had been amended with biosolids which exceeded Cd levels presently allowed. Conversion of land use for these soils is not allowed, but could be allowed if Cd was removed. When the genotype comparisons are completed, breeding of improved strains will begin. Both yield, timing of leaf loss during flowering, height, ease of harvest, and annual shoot Cd will be considered in selecting parents for the breeding work. 4a What was the single most significant accomplishment this past year? We identified that biosolids high in Fe had low water solubility of phosphate. Questions arose about whether this phosphate remained plant available or had been converted to forms which will not be useful in agriculture. Although adding Fe and Al may be able to reduce the water solubility of soil P and protect surface waters, if the less soluble P cannot be used by plants, the method would be less valuable. An experiment was conducted using soils from long term biosolids amended soils using wheat. All soils had high soil test phosphate, but low water soluble phosphate. Nutrients other than P needed to grow wheat on the soils were added. In all treatments tested, sorbed phosphate supplied adequate P for full yield of the plants. The results indicate that adding Fe or Al to manures or biosolids to reduce P solubility does not prevent plants and mycorrhizae from accessing the insoluble phosphate. 4b List other significant accomplishments, if any. Thlaspi caerulescens grown on rice soils in Thailand grew well in a greenhouse test. A field test of high Cd accumulating southern France genotypes was initiated. Sought parties which need to remove Cd from contaminated or mineralized soils to obtain funding needed to conduct this research and prepared several proposals to text Cd remediation and phytoextraction on soils with high Cd:Zn ratios in addition to the tests of rice soils with geogenic ratio of Cd and Zn (1:100). 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. New project just beginning in FY-2005; earlier accomplishments reported in predecessor project. 6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Worked to communicate methods to combine byproducts rich in Fe, Mn and Al to manures and excessively manured soils to groups which need this technology. Gave invited presentation to regional and national meetings and discussed how risk assessment for other elements must be conducted so that any byproduct which may be used to limit P solubility will not cause other long term risks. In particular, the need to obtain balance between Fe and Mn to prevent induced-Mn deficiency, and between Ca and Mg to prevent future Mg deficiency must be achieved in byproduct utilization planning. Because more farmers are finding they will have less ability to apply manures on their own land due to excessive accumulated P, interest in using byproducts to reduce P solubility has increased. 7. List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). R.L. Chaney presented a lecture "Use and Abuse of 503C: Improved Risk Assessment for Contaminants in Non-Biosolids By-Products." at the Great Lakes Byproduct Management Association Annual Meeting and Conference (Decision Making for Blending By-Products: Putting the Pieces Together.) in Chicago, Dec. 2, 2004. Addressed why the CFR 503 rule should not be applied directly to other byproducts or wastes without consideration of the levels of metal sorbents, and the balance among elements for these amendments because they may be very unlike biosolids (the focus of the CFR 503 Rule).

Impacts
(N/A)

Publications