Source: PURDUE UNIVERSITY submitted to
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
Funding Source
Reporting Frequency
Accession No.
Grant No.
Project No.
Proposal No.
Multistate No.
Program Code
Project Start Date
Oct 1, 2010
Project End Date
Sep 30, 2015
Grant Year
Project Director
Sepulveda, M. S.
Recipient Organization
Performing Department
Forestry & Natural Resources
Non Technical Summary
The results of these studies will provide insights into the effects of low-dose exposure to various agrochemicals on non-target species. State and Federal regulatory agencies may use this information in decisions regarding the registration and use of these and other agrochemicals based on a similar mechanism of toxicity. There is a critical need to develop animal models to indicate the impact of agrochemicals, including endocrine-disrupting compounds, on non-target species. The development of fish models into research and regulatory tools for assessing the impact of an agrochemical on non-target species will be very valuable in protecting fish and wildlife. These models will also be useful to the agrochemical industry in evaluating the toxicity of existing and proposed products.
Animal Health Component
Research Effort Categories

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
Goals / Objectives
Determine adverse impacts from agrochemical exposure to cells, organisms, and ecosystems. This research will investigate the environmental impact of agrochemical exposure to non-target organisms from the cellular level to the population level.
Project Methods
Researchers at Purdue University (Indiana) will apply genomic, proteomic, and metabolomic tools to better understand the mechanisms of toxicity as well as develop biomarkers of exposure for a range of environmental contaminants. The test models will include a fish (fathead minnow, Pimephales promelas) and invertebrates (Hyalella azteca and Daphnia magna). For the microarrays, the researchers will utilize a recently developed array that is targeted towards early life stages of fathead minnow, as well as a microarray for H. azteca recently developed in collaboration with scientists from Indiana University and that interrogates over 500 K reads. Proteomic analyzes will be conducted using two-dimensional gel electrophoresis coupled with MALDI TOF/TOF mass spectrometry. For the metabolomic analyses, the researchers will use a combination of GC-MS and LC-MS.

Progress 10/01/11 to 09/30/12

OUTPUTS: The goals of this project are to: 1. Identify, develop, and/or validate trace residue analytical methods, immunological procedures, and biomarkers of chemical exposure and effects. 2. Characterize abiotic and biotic reaction mechanisms, transformation rates, and fate of chemicals in agricultural and natural ecosystems. 3. Determine adverse impacts from agrochemical exposure to cells, organisms, and ecosystems. 4. Develop technologies that mitigate adverse human and environmental impacts. PARTICIPANTS: Indiviaduals: Jessica Leet, PhD student, FNR, Purdue University Linda Lee, Professor, Agronomy, Purdue University Tomas Hook, Assistant Professor, Purdue University Partner Organizations: USEPA TARGET AUDIENCES: Formal classroom instruction and laboratory instruction for FNR undergraduate students. Scientific and technical presentations to scientists from academia, government and industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Concentrated animal feeding operation (CAFO) waste is a cost effective fertilizer. In the Midwest, a network of subsurface tile-drains facilitates transport of animal hormones and nutrients from land-applied CAFO waste to adjacent waterways. Our objective was to better understand impacts of land-applied CAFO waste on fish populations and communities. Water hormone concentrations were characterized from study sites. Fish assemblage structure, reproductive condition, and potential endocrine disruption were assessed in selected fish species. Although most CAFO water samples had hormone concentrations < 1 ng/L, peaks in 17&#946;-E2 (32.95 ng/L) and 17&#945;-TB (34.28 ng/L) equivalent concentrations were registered when resident fishes spawn, hatch, and develop. CAFO sites had lower fish species richness, and fishes exhibited lower reproductive condition compared to individuals from the reference site. Fathead minnows (Pimephales promelas) exposed to CAFO ditchwater during early developmental stages exhibited significantly skewed sex ratios towards males. Maximum observed hormone loads were well above the lowest observable effect concentrations for these hormones. Decrease species richness, altered gonadal development, increased growth rates, and skewed sex ratios towards males could be a result of hormone exposure. Further research is therefore needed that investigates potential links between hormone loads and aquatic biotic assemblages.


  • Baumgarner, B.L., Riley, C.P., Sepulveda, M.S., Brown, P.B., Meyer, J.L., Adamec J., and Sepulveda, M.S. (2012) Increased expression of GAPDH protein is not indicative of nitrosative stress or apoptosis in liver of starved rainbow trout (Oncorhynchus mykiss), Fish Physiology and Biochemistry, 38:379-398.
  • Maity, S., Jannash, A., Adamec, J., Gribskov, M., Nalepa, T.F, Hook TO, Sepulveda MS. (2012 a) Metabolite profiles in starved Diporeia spp. using liquid chromatography-mass spectrometry (LC-MS) based metabolomics, Journal of Crustacean Biology, 32:239-248.
  • Maity. S., Jannasch, A., Adamec, J., Nalepa, T.F., Hook, T.O., and Sepulveda, M.S. (2012 b) Starvation causes disturbance in amino acid and fatty acid metabolism in Diporeia spp., Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology, 161:348-355.
  • Moraga. P., Kinsella, J.M., and Sepulveda, M.S. (2012) Helminth parasites of Eastern box turtles (Terrapene carolina carolina) from Southern Indiana, USA, Journal of Helminthology, 86:38-40.
  • Nutile, S., Amberg, J.J., Sepulveda, M.S., and Goforth, R.R. (2012) Effects of multiple electrical field exposures on Cyprinid embryo survival, North American Journal of Fisheries Management, 32:875-879.
  • Ryan, D.J., Sepulveda, M.S., Nalepa, T.F., and Hook, T.O. (2012) Spatial variation in RNA:DNA ratios of Diporeia spp. in the Great Lakes region, Journal of Great Lakes Research, 38:187-195.
  • Sepulveda, M.S., Patrick, H.K., and Sutton, T.M. 2012a) A single se lamprey attack causes acute anemia and mortality in lake sturgeon, Journal of Aquatic Animal Health, 24:91-99.
  • Sepulveda, M.S., Sutton, T.M., Patrick, H.K., and Amberg, J.J. 2012b) Blood-chemistry values for shovelnose and lake sturgeon, Journal of Aquatic Animal Health, 24:135-140.

Progress 10/01/10 to 09/30/11

OUTPUTS: Results from our research was disseminated through attendance to scientific national and international meetings (total of 49 meetings which were attended by the PI and/or her graduate and undergraduate students); seminars (7) given by the PI; publications in peer-reviewed journals (16); publications in proceedings (1); book chapters (2); invited lectures (29); and extension events (18). PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Undergraduate and graduate students (PhD and MS) General public Federal and state agencies Scientific community, including other researchers PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

These studies have helped in the development of a set of sensitive and robust biomarkers for the quantification of exposure and effects of environmental contaminants on fish and wildlife. The long-term goal of this research is the utilization of these biological responses as early warning signs of problems and for later application in ecological risk assessment evaluations.


  • Leet J, Gall H, Sepulveda MS. 2011 A review of studies on androgen and estrogen exposure in fish early life stages: effects on gene and hormonal control of sexual differentiation. Journal of Applied Toxicology 31:379-398.
  • Stensberg M, McLamore ES, Porterfield DM, Wei Q, Wei A, Sepulveda MS 2011. Toxicological studies on silver nanoparticles: challenges and opportunities in assessment, monitoring and imaging. Nanomedicine 6:879-898.
  • McLamore ES, Sttensberg M, Sepulveda MS, Zhang W, Banks MK, Porterfield DM. 2011 A self-referencing electrode for real time measurements of ionic silver flux. Sens. Actuat. (B: Chem.) 153:445-452.
  • Johns SM, Denslow ND, Kane MD, Watanabe KH, Orlando EF, Sepulveda MS. 2011 Effects of estrogens and antiestrogens on gene expression of fathead minnow (Pimephales promelas) early life stages. Environmental Toxicology 26:195-206.
  • Ralston-Hooper K, Sanchez BC, Jannash A, Adamec J, Sepulveda MS. 2011 Proteomics in aquatic amphipods: Can it be used to determine mechanisms of toxicity and interspecies responses after exposure to atrazine Environmental Toxicology and Chemistry 30:1197-1203.
  • Sanchez BC, Ralston-Hooper K, Sepulveda MS. 2011 A review of recent proteomic applications in aquatic toxicology. Environmental Toxicology and Chemistry 30:274-282.
  • Li Z, Kroll KJ, Jensen KM, Villeneuve DL, Ankley GT, Brian JV, Sepulveda MS, Orlando EF, Lazorchak JM, Kostich M, Armstrong B, Denslow ND, Watanabe KH. 2011 A computational model of the hypothalamic: pituitary: gonadal axis in female fathead minnows (Pimephales promelas) exposed to 17α-ethynylestradiol and 17β-trenbolone. BMC Systems Biology 5:63. doi: 10.1186/1752-0509-5-63.
  • Ralston-Hooper KJ, Adamec J, Jannash A, Mollenhauer R, Ochoa-Acuna H, Sepulveda MS. 2011 Use of GCxGC/TOF-MS and LC/TOF-MS for metabolomic analysis of Hyalella azteca chronically exposed to atrazine and Its primary metabolite, desethylatrazine. Journal of Applied Toxicology 31:399-410.
  • Carlson-Bremer D, Norton TM, Gilardi KV, Dierenfeld ES, Winn B, Sanders FJ, Cray C, Oliva M, Chen TC, Gibbs SE, Sepulveda MS, Johnson CK. 2010 Health assessment of American oystercatchers (Haematopus palliatus palliatus) in Georgia and South Carolina. Journal of Wildlife Diseases 46:772-780.
  • Sepulveda MS, Stefanavage T, Goforth R. 2010 First record of Polypodium sp. parasitizing eggs from female shovelnose sturgeon from the Wabash River, Indiana. Journal of Aquatic Animal Health 22:36-38.
  • Sanchez BC, Carter BJ, Hammers HR, Sepulveda MS. 2011 Transcriptional response of hepatic largemouth bass (Micropterus salmoides) mRNA upon exposure to environmental contaminants. Journal of Applied Toxicology 31:108-116.
  • Laban G, Nies LF, Turco RF, Bickham JW, Sepulveda MS. 2010 The effects of silver nanoparticles on development of fathead minnow (Pimephales promelas) embryos. Ecotoxicology 19:185-195.
  • Yin-Ming K, Sepulveda MS, Sutton TM, Ochoa-Acuna HG, Muir AM, Miller B, Hua I. 2010 Bioaccumulation and biotransformation of decabromodiphenyl ether in juvenile lake whitefish (Coregonus clupeaformis). Ecotoxicology 19:751-760.
  • Yin-Ming K, Sepulveda MS, Hua I, Ochoa-Acuna HG, Sutton, TM. 2010 Biomagnification and trophic transfer of polybrominated diphenyl ethers (PBDEs) in a food web of Lake Michigan. Ecotoxicology 19:623-634.
  • Sepulveda MS, Ralston-Hooper K, Sanchez BC, Hopf-Jannash A, Baker SD, Diaz N, Adamec J 2010. Use of Proteomics and Metabolomics Techniques in Ecotoxicology. In: Handbook of Systems Toxicology: From Omics Technology to Nanotechnology. Sahu SC and Casciano DA (eds.). John Wiley & Sons Ltd. By Invitation Only.
  • Ralston-Hooper K, Hopf A, Adamec J, Sepulveda MS 2010. The Use of GCxGC/TOF-MS for Metabolomic Analysis of Polar Metabolites. In: Methods in Molecular Biology: Metabolic Profiling. Metz T (ed.). Humana Press. By Invitation Only.
  • McLamore ES, Stensberg M, Yale G, Ochoa-Acuna H, Sepulveda MS, Sun X, Akkus O, Porterfield DM. 2010. An optical imaging technique for monitoring real time changes in morphology within the cell, tissue, or whole organism spatial domain. Proceedings of SPIE-International Society of Optical Engineering, Defense, Security, and Sensing Conference, Orlando, FL. Paper No. 7674-14.