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Assessing Impacts of Land-Applied Wastes from Concentrated Animal Feeding Operations on 4 5 6 Fish Populations and Communities 7 8 9 10 1 2 3 1 2 11 Jessica K

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Assessing Impacts of Land-Applied Wastes from Concentrated Animal Feeding Operations on 4 5 6 Fish Populations and Communities 7 8 9 10 1 2 3 1 2 11 Jessica K Environmental Science & Technology Assessing impacts of land -applied wastes from concentrated animal feeding operations on fish populations and communities Journal: Environmental Science & Technology Manuscript ID: Draft Manuscript Type: Article Date Submitted by the Author: n/a Complete List of Authors: Leet, Jessica; Purdue University, Lee, Linda; Purdue University, Gall, Heather; Purdue University, Goforth, Reuben; Purdue University, Sassman, Stephen Gordon, Denise; US Environmental Protection Agency, Ecological Exposure Research Division, Lazorchak, James; US Environmental Protection Agency, Ecological Exposure Research Division, Smith, Mark; US Environmental Protection Agency, Ecological Exposure Research Division, Jafvert, Chad; Purdue University, Civil Engineering Sepulveda, Maria; Purdue University, Civil Engineering ACS Paragon Plus Environment Page 1 of 35 Environmental Science & Technology 1 2 3 Assessing impacts of land-applied wastes from concentrated animal feeding operations on 4 5 6 fish populations and communities 7 8 9 10 1 2 3 1 2 11 Jessica K. Leet , Linda S. Lee , Heather E. Gall , Reuben R. Goforth , Stephen Sassman , Denise 12 4 4 4 3 1, 3* 13 A. Gordon , James M. Lazorchak , Mark E. Smith , Chad T. Javfert , and Maria S. Sepúlveda 14 15 16 17 1 18 Department of Forestry and Natural Resources, Purdue University, 195 Marsteller Street, West 19 20 Lafayette, IN 47907 21 22 2 Department of Agronomy, Purdue University, 915 W. State Street, West Lafayette, IN 47907 23 24 3 25 School of Civil Engineering, Purdue University, 550 Stadium Mall Drive, Lafayette, IN 47907 26 27 4 US Environmental Protection Agency, Ecological Exposure Research Division, 26 West Martin 28 29 Luther King Drive, Cincinnati, OH 45268 30 31 32 33 34 *Corresponding author: Maria S. Sepúlveda, Purdue University, 195 Marsteller Street, (Phone) 35 36 37 765-496-3428, (Fax) 765-496-2422 (Email) [email protected] 38 39 40 41 Table of Content (TOC) Art 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 1 ACS Paragon Plus Environment Environmental Science & Technology Page 2 of 35 1 2 3 Abstract 4 5 6 Concentrated animal feeding operation (CAFO) waste is a cost effective fertilizer. In the 7 8 Midwest, networks of subsurface tile-drains expedite transport of animal hormones and nutrients 9 10 11 from land-applied CAFO waste to adjacent waterways. The objective of this study was to 12 13 evaluate impacts of land-applied CAFO waste on fish populations and communities. Water 14 15 chemistry including hormone, pesticide, and nutrient concentrations was characterized from 16 17 18 study sites along with fish assemblage structure, growth, and endocrine disruption were assessed 19 20 in selected fish species. Although most CAFO water samples had hormone concentrations < 1 21 22 ng/L, equivalent concentrations for 17 β-E2 and 17 α-TB peaked at > 30 ng/L each during the 23 24 25 period of spawning, hatching, and development for resident fishes. CAFO sites had lower fish 26 27 species richness, and fishes exhibited faster somatic growth and lower reproductive condition 28 29 compared to individuals from the reference site. Fathead minnows ( Pimephales promelas ) 30 31 32 exposed to CAFO ditchwater during early developmental stages exhibited significantly skewed 33 34 sex ratios towards males. Maximum observed hormone concentrations were well above the 35 36 37 lowest observable effect concentrations for these hormones; however, complexities at the field 38 39 scale make it difficult to directly relate hormone concentration and impacts on fish. 40 41 Complicating factors include the consistent presence of pesticides and nutrients, and the 42 43 44 difference in temperature and stream architecture of the CAFO-impacted ditches compared to the 45 46 reference site (e.g., channelization, bottom substrate, shallow pools, and riparian cover). 47 48 49 50 51 52 53 54 55 56 57 58 59 60 2 ACS Paragon Plus Environment Page 3 of 35 Environmental Science & Technology 1 2 3 Introduction 4 5 6 Wastes from concentrated animal feeding operations (CAFOs) are often applied to 7 8 agricultural fields as a waste management strategy and a source of inexpensive fertilizer and/or 9 10 11 irrigation water. Subsurface tile-drain networks are widely used in the Midwestern U.S. to lower 12 13 the water table in agricultural fields. However, these networks also expedite the transport of 14 15 nutrients, pesticides, and manure-borne constituents to receiving ditches and downstream 16 17 1 2, 3 18 waterways. Manure can transfer synthetic and natural hormones to the environment. Timing 19 20 is critical for organisms exposed to hormones. Thus, even though hormones have short half-lives 21 22 (days; 4), brief exposures during critical developmental periods can induce long-lasting effects.5 23 24 25 Few studies have investigated the effects of CAFO-impacted water on fish reproduction. 26 27 Sellin et al. 6, 7 observed a decrease in estrogen responsive genes in caged fathead minnows 28 29 (Pimephales promelas ) deployed for 7 d in a stream close to a beef CAFO. The only published 30 31 32 field study of feral fish inhabiting streams receiving feedlot effluent demonstrated reduced 33 34 reproductive fitness in male fathead minnows. 8 These studies suggest that fishes are affected by 35 36 37 CAFO-impacted waters, although more explicit studies are needed to assess the potential 38 39 biological and ecological impacts of land-applied CAFO waste. 40 41 Our objective was to further evaluate the impacts of land-applied CAFO waste on fishes. 42 43 44 We hypothesize that sites near fields receiving CAFO wastes have greater hormone and nutrient 45 46 concentrations, lowered fish species richness and evidence of endocrine disruption, compared to 47 48 a reference site. To test these hypotheses, we evaluated water quality including hormone, 49 50 51 pesticide, and nutrient concentrations, fish abundance and species richness, and growth and 52 53 reproductive characteristics of feral creek chub (Semotilus atromaculatus ). Finally, we 54 55 56 57 58 59 60 3 ACS Paragon Plus Environment Environmental Science & Technology Page 4 of 35 1 2 3 conducted in situ exposures of fathead minnow embryos and caged adults to CAFO-impacted 4 5 6 waters. 7 8 9 10 11 Materials and Methods 12 13 Study sites 14 15 Studies were conducted at two CAFO-impacted sites (Marshall and Box Ditches) and one 16 17 18 reference site (Ghost Creek). Ditches receive subsurface tile drainage and runoff from adjacent 19 20 agricultural fields located at Purdue University’s Animal Science Research and Education 21 22 Center, West Lafayette, IN (Figure 1). CAFO wastes are land-applied via solid broadcasting, 23 24 1 25 pivot irrigation, or subsurface injection (see Gall et al. for details). Ghost Creek is a tributary to 26 27 the Tippecanoe River ~ 25 km northeast of the CAFO sites near Brookston, IN (Figure 1). 28 29 Marshall and Box Ditches are channelized agricultural streams with extensive bank 30 31 32 erosion and sedimentation, few pools, and no riffle habitats. Base flow is low, and only a few 33 34 shallow pools persist during late summer and early fall. Only one riparian canopy was present 35 36 37 (transect M2). In contrast, Ghost Creek is a meandering stream surrounded by forest with 38 39 riparian canopy throughout. Aside from a stone path used to cross the stream at transect G2, 40 41 Ghost Creek has little bank erosion and sedimentation. Each Ghost Creek transect contained 42 43 44 several pool-riffle sequences characterized by substrates ranging from sand to cobbles. 45 46 47 48 Water quality and chemical analysis 49 50 51 Temperature (°C) and dissolved oxygen (DO, mg/L) were measured using YSI meters 52 53 (Yellow Springs, OH, USA). Water for chemical analysis was collected by automated sampling 54 55 stations S1, S2, and S3 (Figure 1). These sites corresponded to Marshall Ditch transects M3 and 56 57 58 59 60 4 ACS Paragon Plus Environment Page 5 of 35 Environmental Science & Technology 1 2 3 M2 and Box Ditch transect B3, respectively. Hormones and pesticides were quantified using 4 5 6 solid phase extraction, eluting with methanol, evaporating eluant and reconstituted residues in 7 8 methanol (0.5 mL) followed by high performance reverse-phase liquid chromatography tandem 9 10 11 electrospray ionization mass spectrometry (HPLC/ESI-MS/MS) for the hormones (details 12 1 13 provided in Gall et al. ) and GC/MS for the pesticides. Hormones included estrone (E1), 17 α- 14 15 estradiol (17 α-E2), 17 β-estradiol (17 β-E2), estriol (E3), testosterone (TST), androstenedione 16 17 18 (AND), 17 α-trenbolone (17 α-TB), 17 β-trenbolone (17 β-TB), and trendione (TND). Pesticides 19 20 included atrazine, deethyl-atrazine (DEA), deisopropyl-atrazine (DIA), and acetochlor. USEPA 21 22 - - 3- Methods 353.2 and 365.1 to analyze NO 3 -N + NO 2 -N and PO 4 -P with a Seal AQ2+ Advanced 23 24 25 Discrete Analyzer. Total nitrogen (TN) and total phosphorus (TP) were determined from grab 26 27 samples collected at all three sites during creek chub sampling by TN–persulfate (method 10072) 28 29 and TP–PhosVer®3 with acid persulfate digestion (method 8190) kits (Hach spectrophotometer, 30 31 32 DR 2700, Loveland, CO, USA). 33 34 Maximum observed hormone values were converted to equivalency values and summed 35 36 37 to determine total estrogen and androgen potency at each site. Data for relative binding to 38 39 fathead minnow estrogen (ER) and androgen (AR) receptors were used in these calculations 40 41 because this species is closely related to cyprinids found at the study sites. Estrogen equivalency 42 43 44 values were calculated based on binding of each estrogen relative to the binding affinity of 17 β- 45 9 46 E2 to fathead minnow ER, since 17 β-E2 was the most potent estrogen detected (Table 1).
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