Funding for the work described here came from the California Department of Pesticide Regulation and the California Department of Water Resources.

1. Urban Bioswale Efficacy Monitoring
<ol>
	<li>Storm Water Sampling
	<ol>
		<li>Sample 4 L of pre-treatment stormwater leaving the parking lot as it enters the bioswale inlet, and then 4 L of post-treatment stormwater as it leaves the bioswale through the 4&#34; outlet drain.</li>
		<li>Using local weather predictions, collect samples at the beginning, middle, and end of the storm&#39;s hydrograph. Composite the samples to characterize runoff variability during the storm event.</li>
		<li>Collect 1.3 L samples by hand and composite them in a 4 L amber bottle. Collect inlet samples at several curb openings where storm water flows into the bioswale.</li>
		<li>Collect 1.3 L outlet samples from the flow meter attached to the outlet drain (described below) and composite them in a 4 L amber bottle.</li>
		<li>Store the composited samples on ice until the final hydrograph sample is collected. Then transport them to the laboratory and hold in a refrigerator at 4 &#176;C prior to subsampling for chemistry and toxicity testing. Ship samples to the chemistry lab within 48 h of sample collection.</li>
	</ol>
	</li>
	<li>Load Calculation
	<ol>
		<li>Prior to the storm, install a tipping-bucket digital logger rain gauge by attaching it to a light or other pole adjacent to the bioswale site. Use the rain data to indicate instantaneous and total rainfall for the site.</li>
		<li>Install a mechanical geared pulse flow meter on the outlet drains of the bioswale. Record total flow exiting the bioswale. 
		NOTE: Reduction in runoff volume is presumed to reduce overall loading of contaminants in LID designs.</li>
		<li>Model the volume of water falling on the parking lot catchment area during the rain event by extrapolation using the inches of rain recorded by the rain gauge. Use these data to determine the volume entering the treatment system based on parking lot surface area.</li>
		<li>Use the total flow recorded by the outlet flow meter to calculate infiltration percentage. Calculate the difference between inlet and outlet volume to determine stormwater infiltration.</li>
		<li>Calculate contaminant loading and load reduction percentages during the storm using inlet and outlet volume in conjunction with contaminant analytical measurements.</li>
		<li>Measure chemical analytes that are relevant to surface water toxicity (as discussed below). Total chemical groups to simplify load calculations and base on their similar toxic modes of action (e.g., total polynuclear aromatic hydrocarbons [PAHs], total pyrethroids, and total fipronil and degradates).</li>
	</ol>
	</li>
	<li>Chemistry
	<ol>
		<li>Analyze all samples for the following parameters: total suspended solids (TSS), trace metals (USEPA method 200.85; inductively coupled plasma-mass spectrometry [ICP/MS]), and PAHs (USEPA method 6256).</li>
		<li>Analyze samples for current-use urban pesticides, including 9 pyrethroids (USEPA method SW846 8270 modified7; bifenthrin, cypermethrin, fenvalerate/esfenvalerate, permethrin, tetramethrin, L-cyhalothrin, cyfluthrin, and allethrin), and fipronil and its three primary degradates (fipronil sulfide, fipronil sulfone, fipronil desulfinyl).</li>
		<li>Analyze pyrethroids using gas chromatography-mass spectrometry (GC/MS) using negative chemical ionization or other appropriate method to provide adequate detection limits. Since most current-use pesticides are highly toxic at low concentrations, their analyses require low chemical reporting limits to be relevant for environmental risk assessment. The method reporting limits for pyrethroids are from 0.5 ng/L to 1.0 ng/L for all pyrethroids except permethrin (reporting limit = 10 ng/L).</li>
		<li>Use an analytical procedure for fipronil that provides a method reporting limit of 1.0 ng/L. Organophosphate pesticides do not need to be measured depending on local use patterns for example in urban areas in California8,9.</li>
		<li>Measure neonicotinoid pesticides (e.g., imidacloprid) using ultra performance liquid chromatography coupled to a triple quadrupole mass spectrometer, which has a reporting limit for imidacloprid of 50 ng/L.</li>
	</ol>
	</li>
	<li>Toxicity Testing
	<ol>
		<li>Conduct toxicity tests on the composited inlet and outlet stormwater samples using 3 test species, following modified US Environmental Protection Agency (USEPA) acute test protocols10. The test with the cladoceran Ceriodaphnia dubia measures survival after 96 h. The test with the amphipod Hyalella azteca measures survival after 10 days. The test with the midge Chironomus dilutus measures survival and growth after 10 days.</li>
		<li>Conduct acute 96 h survival tests with the cladoceran C. dubia following U.S. EPA guidance.
		<ol>
			<li>Expose five C. dubia neonates in each of five replicates of inlet and outlet stormwater samples. Replicates consist of 20-mL scintillation vials containing 15 mL of test solution.</li>
			<li>Feed neonates a mixture of yeast, cerophyll, trout chow (=YCT; following U.S. EPA guidance) and Selenastrum algae 2 h prior to daily 100% renewal of the stormwater test solutions. Record total number of surviving neonates daily.</li>
			<li>Compare final C. dubia survival after 96 h exposure to inlet and outlet stormwater samples to survival in moderately hard control water using a t-test. Follow statistical procedures recommended by U.S. EPA.</li>
		</ol>
		</li>
		<li>Conduct acute 10 d survival tests with the amphipod H. azteca following U.S. EPA guidance.
		<ol>
			<li>Expose 10, 9 days to 15 days old amphipods in each of five replicates. Replicates consist of 300-mL glass beakers containing 200 mL of test solution.</li>
			<li>Conduct amphipod tests for 10 days, count the number of surviving amphipods daily, and renew 50% of the test solution every 48 h. Feed each beaker every 48 h with 1.5 mL of YCT after the renewal.</li>
			<li>Compare final survival of amphipods in stormwater samples to 10 days survival in laboratory well water as described above.</li>
		</ol>
		</li>
		<li>Conduct chronic 10 d survival and growth tests with the midge C. dilutus following U.S. EPA guidance.
		<ol>
			<li>Expose 12, 7-d old animals in each of four replicates. Replicates consist of 300 mL glass beakers containing 200 mL of test solution. Supply each midge test container with 5 mL of sand as substrate for tube building by the larvae.</li>
			<li>Conduct tests for 10 d, and renew 50% of the test solution every 48 h for each beaker daily with an increasing amount of fish food slurry (4 g/L), as follows: days 0 to 3, 0.5 mL/day; days 4 to 6, 1.0 mL/day; days 7 to 10, 1.5 mL/day.</li>
			<li>Compare final survival and growth in stormwater samples to 10-d survival in laboratory well water as described above. Measure growth of surviving animals as ash-free dry weight at 10 d compared to initial weight of the test organisms.</li>
		</ol>
		</li>
		<li>For all toxicity tests, measure dissolved oxygen, pH, and conductivity using appropriate meters and electrodes. Measure un-ionized ammonia using a spectrophotometer.
		<ol>
			<li>Measure water hardness and alkalinity at initiation and termination of tests.10</li>
			<li>Record water temperature with a continuous recording thermometer.</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>
2. Integrated Vegetated Agricultural Drainage Ditch Efficacy Monitoring
<ol>
	<li>Integrated Ditch Construction 
	NOTE: The agricultural drainage ditch used in the current example is 152 m long and has a semi-V-shaped cross-section width of 5 m at the top and 1 m depth. The ditch vegetation is a combination of native grass species primarily seeded with red fescue (Festuca rubra). In this example, integrated vegetative ditch trials consisted of granulated activated carbon (GAC) and compost filter treatments integrated with the vegetated ditch.
	<ol>
		<li>Construct two compost filters and six carbon filters and install them in three different sections of the vegetated ditch (Figure 2). Use 2 m long 20 cm diameter sleeves filled with either carbon or compost.</li>
		<li>Fill six sleeves with 30 L of granulated activated carbon and place these across the ditch at the 146 m point, near the end of 152 m vegetated ditch. Anchor the GAC-filled sleeves to the ditch bottom with wire stakes on the upstream edge.</li>
		<li>Place a 2.5 m long 6&#34; wide section of pine board on the downstream edge of each of the GAC sleeves. Dig the pine boards into the two sides and bottom of the channel to minimize water bypassing and undercutting the carbon sleeves. The boards will also provide vertical support to maximize water contact time with the carbon.</li>
		<li>Fill the compost sleeves with approximately 15 kg each of partially decomposed yard waste from any clean source, such as a local landfill. Position two 2 m long compost sleeves across the vegetated channel at 64 m and at 123 m along the length of the 152 m vegetated ditch (Figure 2).</li>
	</ol>
	</li>
	<li>Runoff Simulation and Sampling 
	NOTE: This protocol describes methods for conducting simulated agricultural runoff trials and associated monitoring to evaluate treatment efficacy using the integrated vegetative treatment system. In the current example, the integrated vegetated-compost-carbon system was evaluated at two flow rates that represented rates typical off-field discharge from commercial farms in the Salinas Valley, 3.2 L/s and 6.3 L/s. The organophosphate pesticide chlorpyrifos was used as a model pesticide in these trials because it has a moderate solubility, and therefore represents the mid-range of solubility of representative pesticides commonly used in pest management. Chlorpyrifos is also the subject of on-going regulatory actions in central California because of its impacts on agriculture watersheds. The target chlorpyrifos dose was approximately 2,600 ng/L. Flow rates and target chlorpyrifos concentrations were within the ranges previously measured in local irrigation runoff 3,11. The hydraulic residence time for a pulse of water transiting the vegetated ditch was not monitored in the example given here. The residence time in these systems vary with water inflow rate, the degree of soil saturation due to previous irrigation and rain, the presence of structures to impede flow such as weirs and sedimentation basins, and the amount of surface area covered by vegetation. Previous studies have demonstrated residence times of several hours for small scale ditch systems in the Salinas Valley 3,4. Visual observations indicated the residence time for the GAC filters was one or two minutes.
	<ol>
		<li>Create simulated agricultural runoff using ground water mixed with suspended sediment. For trials with the model pesticide, chlorpyrifos, prepare a fresh stock solution of 10 mg/L for every 3.2 L/s trial by adding certified stock solution to a known volume of distilled water. Prepare a fresh chlorpyrifos stock solution of 20 mg/L for every 6.3 L/s trial.
		<ol>
			<li>Use a metering pump to provide a consistent volume of stock solution to the runoff water before it enters the vegetated treatment ditch inlet. Use the metering pump to deliver stock solution at 50 mL/min to the flow of simulated irrigation water.</li>
		</ol>
		</li>
		<li>Monitor the inlet flow rate with a digital meter and use these data to quantify total volume of runoff water applied to the ditch inlet.</li>
		<li>Construct a weir at the outlet of the ditch and plumb this with an outlet pipe connected to a digital flow meter. Use this meter to record the volume of runoff exiting the ditch.</li>
		<li>Use data loggers connected to the digital meters to record flow at 5 min intervals. Program the data loggers to activate peristaltic pumps located at the inlet and at various stations (e.g., 23 m, 45 m, and 68 m) below the inlet of the ditch to collect composite subsamples of runoff into stainless steel containers at 5 min intervals.</li>
	</ol>
	</li>
	<li>Chemistry
	<ol>
		<li>Transfer composite samples of runoff water from trials into amber glass bottles at the end of each runoff trial and maintain the samples on ice at 4 &#176;C for later toxicity and chemical analyses.</li>
		<li>Analyze the composite samples for total suspended solids (TSS), and chlorpyrifos using GC-MS or enzyme-linked immunosorbent assays (ELISA).</li>
		<li>Compare &#34;inlet&#34; composite samples (pre-treatment) to &#34;outlet&#34; composite samples (post-treatment) to evaluate efficacy of the integrated ditch system to reduce TSS and pesticide loads.</li>
	</ol>
	</li>
	<li>Toxicity Testing
	<ol>
		<li>Determine water column toxicity was in composite samples from the inlet (pre-treatment) and outlet (post-treatment) of each trial using 96 h Ceriodaphnia dubia toxicity tests10, as described above for the bioswale monitoring. C. dubia is an appropriate monitoring species for agriculture runoff toxicity due to its sensitivity to chlorpyrifos (median lethal concentration (LC50) = 53 ng/L12).</li>
	</ol>
	</li>
</ol>

<ol>
	<li>Anderson, B. S., Hunt, J. W., Markewicz, D., Larsen, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Toxicity in California Waters, Surface Water Ambient Monitoring Program. , (2011).</li><li>Anderson, B. S., Phillips, B. M., Voorhees, J. P., Siegler, K., Tjeerdema, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioswales+reduce+contaminants+associated+with+toxicity+in+urban+stormwater.">Bioswales reduce contaminants associated with toxicity in urban stormwater.</a> Environ Toxicol Chem. 35 (12), 3124-3134 (2016).</li><li>Anderson, B. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pesticide+and+toxicity+reduction+using+an+integrated+vegetated+treatment+system.">Pesticide and toxicity reduction using an integrated vegetated treatment system.</a> Environ Toxicol Chem. (30), 1036-1043 (2011).</li><li>Phillips, B. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mitigation Strategies for Reducing Aquatic Toxicity from Chlorpyrifos in Cole Crop Irrigation Runoff. , (2014).</li><li>U.S. EPA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Method 1640: Determination of Trace Elements in Ambient Waters by On-Line Chelation Pre-concentration and Inductively Coupled Plasma-Mass Spectrometry. , (1995).</li><li>U.S. EPA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods for organic chemical analysis of municipal and industrial wastetwater, Method 625- Base/neutrals and acids. , (1984).</li><li>U.S. EPA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> , (1993).</li><li>Johnson, H. M., Domagalski, J. L., Saleh, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trends+in+Pesticide+Concentrations+in+Streams+of+the+Western+United+States.">Trends in Pesticide Concentrations in Streams of the Western United States.</a> J Am Water Resour Assoc. 47 (2), 265-286 (1993).</li><li>Siegler, K., Phillips, B. M., Anderson, B. S., Voorhees, J. P., Tjeerdema, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+and+spatial+trends+in+sediment+contaminants+associated+with+toxicity+in+California+watersheds.">Temporal and spatial trends in sediment contaminants associated with toxicity in California watersheds.</a> Environ Poll. , 1-6 (2015).</li><li>U.S. EPA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods for measuring acute toxicity of effluents and receiving water to freshwater and marine organisms. , (2002).</li><li>Bailey, H. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Joint+acute+toxicity+of+diazinon+and+chlorpyrifos+to+Ceriodaphnia+dubia.">Joint acute toxicity of diazinon and chlorpyrifos to Ceriodaphnia dubia.</a> Environ Toxicol Chem. 16, 2304-2308 (1997).</li><li>Supowit, S., Sadaria, A. M., Reyes, E. J., Halden, R. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mass+balance+of+fipronil+and+total+toxicity+of+fipronil-related+compounds+in+process+streams+during+conventional+wastewater+and+wetland+treatment.">Mass balance of fipronil and total toxicity of fipronil-related compounds in process streams during conventional wastewater and wetland treatment.</a> Environ Sci Technol. 50 (3), 1519-1526 (2016).</li><li>Stang, C., Bakanov, N., Schulz, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experiments+in+water-macrophyte+systems+to+uncover+the+dynamics+of+pesticide+mitigation+processes+in+vegetated+surface+waters/streams.">Experiments in water-macrophyte systems to uncover the dynamics of pesticide mitigation processes in vegetated surface waters/streams.</a> Environ Sci Pollut Res. , (2015).</li><li>Schulz, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Field+studies+on+exposure,+effects,+and+risk+mitigation+of+aquatic+nonpoint-source+insecticide+pollution:+A+review.">Field studies on exposure, effects, and risk mitigation of aquatic nonpoint-source insecticide pollution: A review.</a> J Environ Qual. 33 (2), 419-448 (2004).</li><li>Moore, M. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transport+and+fate+of+atrazine+and+lambda-cyhalothrin+in+a+vegetated+drainage+ditch+in+the+Mississippi+Delta.">Transport and fate of atrazine and lambda-cyhalothrin in a vegetated drainage ditch in the Mississippi Delta.</a> Agric Ecosyst Environ. 87, 309-314 (2001).</li><li>Phillips, B. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effects+of+the+Landguard+A900+Enzyme+on+the+Macroinvertebrate+Community+in+the+Salinas+River,+California,+United+States+of+America.">The Effects of the Landguard A900 Enzyme on the Macroinvertebrate Community in the Salinas River, California, United States of America.</a> Arch Environ Contam Toxicol. 70 (2), 231-240 (2016).</li><li>Han, W., Fang, J., Liu, X., Tang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Techno-economic+feasibility+evaluation+of+a+combined+bioprocess+for+fermentative+hydrogen+production+from+food+waste.">Techno-economic feasibility evaluation of a combined bioprocess for fermentative hydrogen production from food waste.</a> Bioresource Technology. , 107-112 (2016).</li><li>Solomon, K. R., Giddings, J. M., Maund, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probabilistic+risk+assessment+of+cotton+pyrethroids:+I.+Distributional+analysis+of+laboratory+aquatic+toxicity+data.">Probabilistic risk assessment of cotton pyrethroids: I. Distributional analysis of laboratory aquatic toxicity data.</a> Environ Toxicol Chem. 20, 652-659 (2001).</li><li>Weston, D. P., Lydy, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toxicity+of+the+Insecticide+Fipronil+and+Its+Degradates+to+Benthic+Macroinvertebrates+of+Urban+Streams.">Toxicity of the Insecticide Fipronil and Its Degradates to Benthic Macroinvertebrates of Urban Streams.</a> Environ Sci Tech. , (2014).</li><li>Voorhees, J. P., Anderson, B. S., Phillips, B. M., Tjeerdema, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carbon+treatment+as+a+method+to+remove+imidacloprid+from+agriculture+runoff.">Carbon treatment as a method to remove imidacloprid from agriculture runoff.</a> Bull Environ Contam Toxicol. , (2017).</li></ol>

The authors declare that they have no competing financial interests.

The practices described in this protocol are intended as final steps in an overall strategy to remove pollutants in agriculture irrigation and stormwater runoff. Use of bioswales and other urban green-infrastructure LID practices are intended as a final piece of the puzzle to remove contaminants in runoff before they reach adjacent receiving waters. This protocol emphasizes methods to monitor urban bioswales to determine treatment efficacy for removing toxicity associated with urban contaminants, with emphasis on current...

Surface water toxicity is prevalent in California watersheds and decades of monitoring have shown that toxicity is often due to pesticides and other contaminants1. The primary sources of surface water contamination are stormwater and irrigation runoff from urban and agriculture sources. As waterbodies are listed as degraded due to contaminants and the toxicity is identified from urban and agricultural sources, water quality regulators partner with state and federal funding sources to implement practices to reduce contaminant loading. Green infrastructure is being promoted in California urban watersheds to reduce flooding and increase the recovery of stormwater through infiltration and storage. While Low Impact Development (LID) designs are being mandated for new construction in many regions, few studies have monitored the efficacy of these systems beyond measurements of conventional contaminants like dissolved solids, metals, and hydrocarbons. More intensive monitoring has recently evaluated reductions in chemical concentrations and chemical loading responsible for surface water toxicity, and to directly determine whether bioswales reduce toxicity of runoff. This has shown that bioswales are effective at removing toxicity associated with some contaminant classes2, but additional research is required for emerging chemicals of concern.
Vegetated treatment systems are also being implemented in agriculture watersheds in California, and these have been shown to be effective at reducing pesticides and other contaminants in agriculture irrigation runoff3,4. These systems represent components of a suite of approaches to reduce contaminant loading to surface waters. Because they are intended to mitigate contaminants responsible for surface water toxicity, a key component of the implementation process is monitoring to ensure their long-term effectiveness. Monitoring includes both chemical analyses of chemicals of concern, as well as toxicity testing with sensitive indicator species. This article describes protocols and monitoring results for an urban parking lot bioswale and an agricultural vegetated drainage ditch system.
The design attributes of a typical parking lot bioswale, such as may be used to treat storm runoff in a typical mixed-use urban shopping parking area depend on the area being treated. In the example described here, 53,286 square feet of asphalt create an impervious surface area that drains to a swale, which consists of 4,683 square feet of landscaping. To accommodate runoff from this surface area, a 215 feet long flat-bottom, semi-V shape channel comprises the swale with a side slope less than 50% and a longitudinal slope of 1% (Figure 1). This swale comprises three layers including native bunch grass planted in 6 inches of topsoil, layered over 2.5 feet of compacted subgrade. Stormwater flows from parking areas to multiple entry points along the swale. The water infiltrates the vegetated area, then permeates the subgrade and drains into a 4-inch perforated drain. This system drains water through a system plumbed to an adjacent wetland that eventually drains into a local creek.

<table><tbody><tr><td>HOBO tipping-bucket digital logger rain gauge&amp;nbsp;</td><td>Onset Computer Co., Bourne MA, USA)</td><td>Onset RG3</td><td>Rain gauge</td></tr><tr><td>Mechanical geared pulse flow meter&amp;nbsp;</td><td>Seametrics Inc., Kent WA</td><td>Seametrics MJ-R</td><td>Flow meter for measuring bioswale outlet flow</td></tr><tr><td>Filtrexx SafteySoxx</td><td>Filtrexx Co. - info@filtrexx.com</td><td>SafetySoxx</td><td>perforated synthetic cloth for granulated activated carbon and compost</td></tr><tr><td>Granulated activated carbon&amp;nbsp;</td><td>Evoqua - Siemens Corp., Oakland CA</td><td>AC380</td><td>GAC for agriculture irrigation water treatment</td></tr><tr><td>Digital flow meters&amp;nbsp;</td><td>Seametrics Inc. Kent WA</td><td>Ag2000; WMP101</td><td>Flow meters for agriculture irrigation treatment system monitoring</td></tr><tr><td>Data Loggers</td><td>Campbell Scientific Inc., Logan, UT</td><td>CR1000</td><td>Data loggers for recording flow data</td></tr><tr><td>Peristaltic pumps for composite sampling</td><td>Omega Engineering Inc. Stamford CT</td><td>Omegaflex FPU-122-12VDC&amp;nbsp;</td><td>Pumps for composite sampling</td></tr></tbody></table>

vegetated treatment systems for removing contaminants associated with surface water toxicity in agriculture and urban runoff

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may be treated with simple systems designed to promote sorption of contaminants to vegetation and soils and promote infiltration. Two example systems are described: a bioswale treatment system for urban stormwater treatment, and a vegetated drainage ditch for treating agriculture irrigation runoff. Both have similar attributes that reduce contaminant loading in runoff: vegetation that results in sorption of the contaminants to the soil and plant surfaces, and water infiltration. These systems may also include the integration of granulated activated carbon as a polishing step to remove residual contaminants. Implementation of these systems in agriculture and urban watersheds requires system monitoring to verify treatment efficacy. This includes chemical monitoring for specific contaminants responsible for toxicity. The current paper emphasizes monitoring of current use pesticides since these are responsible for surface water toxicity to aquatic invertebrates.

Urban Bioswale Efficacy
During the 18.5 h of the storm, 1.52&#34; of rain was recorded by the rain gauge, and this resulted in 50,490 gallons of water flowing from the parking lots into the bioswale. Of this total volume, 5,248 gallons were recorded by the outlet flow meter, resulting in a total infiltration of 90% of the stormwater that flowed into the bioswale. The bioswale reduced all of the chemicals monitored. Total suspended ...

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Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

This article summarizes the design attributes and the effectiveness of treatment systems that treat urban stormwater and agriculture irrigation runoff to remove pesticides and other contaminants associated with aquatic toxicity.

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may ...

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10.5K Views.  University of California, Davis. This article summarizes the design attributes and the effectiveness of treatment systems that treat urban stormwater and agriculture irrigation runoff to remove pesticides and other contaminants associated with aquatic toxicity.

Video: Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

Design and Construction of an Urban Runoff Research Facility

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use due to increased demand for landscape irrigation. Improper irrigation practices and large rainfall events can result in runoff from urban landscapes which has potential to carry nutrients and sediments into local streams and lakes where they may contribute to eutrophication. A 1,000 m2 facility was constructed which consists of 24 individual 33.6 m2 field plots, each equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated urban landscapes. Runoff volumes from the first and second trials had coefficient of variability (CV) values of 38.2 and 28.7%, respectively. CV values for runoff pH, EC, and Na concentration for both trials were all under 10%. Concentrations of DOC, TDN, DON, PO4-P, K+, Mg2+, and Ca2+ had CV values less than 50% in both trials. Overall, the results of testing performed after sod installation at the facility indicated good uniformity between plots for runoff volumes and chemical constituents. The large plot size is sufficient to include much of the natural variability and therefore provides better simulation of urban landscape ecosystems.

This paper describes the design, construction, and function of a 1,000 m2 facility containing 24 individual 33.6 m2 field plots equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated home lawns.

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use ...

Integrated Field Lysimetry and Porewater Sampling for Evaluation of Chemical Mobility in Soils and Established Vegetation

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals is often not well understood. Here we demonstrate an integrated field lysimetry and porewater sampling method for evaluating the mobility of chemicals applied to soils and established vegetation. Lysimeters, open columns made of metal or plastic, are driven into bareground or vegetated soils. Porewater samplers, which are commercially available and use vacuum to collect percolating soil water, are installed at predetermined depths within the lysimeters. At prearranged times following chemical application to experimental plots, porewater is collected, and lysimeters, containing soil and vegetation, are exhumed. By analyzing chemical concentrations in the lysimeter soil, vegetation, and porewater, downward leaching rates, soil retention capacities, and plant uptake for the chemical of interest may be quantified. Because field lysimetry and porewater sampling are conducted under natural environmental conditions and with minimal soil disturbance, derived results project real-case scenarios and provide valuable information for chemical management. As chemicals are increasingly applied to land worldwide, the described techniques may be utilized to determine whether applied chemicals pose adverse effects to human health or the environment.

Field lysimetry and porewater sampling allow researchers to evaluate the fate of chemicals applied to soils and established vegetation. The goal of this protocol is to demonstrate how to install required instrumentation and collect samples for chemical analysis during integrated field lysimetry and porewater sampling experiments.

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals ...

Protocols for Robust Herbicide Resistance Testing in Different Weed Species

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented protocols, based on whole-plant bioassays performed in a greenhouse, can be readily adapted to a wide range of weed species and herbicides through appropriate variants. Seed samples from plants that survived a field herbicide treatment are collected and stored dry at low temperature until used. Germination methods differ according to weed species and seed dormancy type. Seedlings at similar growth stage are transplanted and maintained in the greenhouse under appropriate conditions until plants have reached the right growth stage for herbicide treatment. Accuracy is required to prepare the herbicide solution to avoid unverifiable mistakes. Other critical steps such as the application volume and spray speed are also evaluated. The advantages of this protocol, compared to others based on whole plant bioassays using one herbicide dose, are related to the higher reliability and the possibility of inferring the resistance level. Quicker and less expensive in vivo or in vitro diagnostic screening tests have been proposed (Petri dish bioassays, spectrophotometric tests), but they provide only qualitative information and their widespread use is hindered by the laborious set-up that some species may require. For routine resistance testing, the proposed whole plant bioassay can be applied at only one herbicide dose, so reducing the costs.

A robust and flexible approach to confirm herbicide resistance in weed populations is presented. This protocol allows the herbicide resistance levels to be inferred and applied to a wide range of weed species and herbicides with minor adaptations.

Robust protocols to test putative herbicide resistant weed populations at whole plant level are essential to confirm the resistance status. The presented ...

Development of Sulfidogenic Sludge from Marine Sediments and Trichloroethylene Reduction in an Upflow Anaerobic Sludge Blanket Reactor

The importance of microbial sulfate reduction relies on the various applications that it offers in environmental biotechnology. Engineered sulfate reduction is used in industrial wastewater treatment to remove large concentrations of sulfate along with the chemical oxygen demand (COD) and heavy metals. The most common approach to the process is with anaerobic bioreactors in which sulfidogenic sludge is obtained through adaptation of predominantly methanogenic granular sludge to sulfidogenesis. This process may take a long time and does not always eliminate the competition for substrate due to the presence of methanogens in the sludge. In this work, we propose a novel approach to obtain sulfidogenic sludge in which hydrothermal vents sediments are the original source of microorganisms. The microbial community developed in the presence of sulfate and volatile fatty acids is wide enough to sustain sulfate reduction over a long period of time without exhibiting inhibition due to sulfide. 
This protocol describes the procedure to generate the sludge from the sediments in an upflow anaerobic sludge blanket (UASB) type of reactor. Furthermore, the protocol presents the procedure to demonstrate the capability of the sludge to remove by reductive dechlorination a model of a highly toxic organic pollutant such as trichloroethylene (TCE). The protocol is divided in three stages: (1) the formation of the sludge and the determination of its sulfate reducing activity in the UASB, (2) the experiment to remove the TCE by the sludge, and (3) the identification of microorganisms in the sludge after the TCE reduction. Although in this case the sediments were taken from a site located in Mexico, the generation of a sulfidogenic sludge by using this procedure may work if a different source of sediments is taken since marine sediments are a natural pool of microorganisms that may be enriched in sulfate reducing bacteria.

Microbial sulfate reduction is a process of great importance in environmental biotechnology. The success of the sulfidogenic reactors depends among other factors on the microbial composition of the sludge. Here, we present a protocol to develop sulfidogenic sludge from hydrothermal vents sediments in a UASB reactor for reductive dechlorination purposes.

The importance of microbial sulfate reduction relies on the various applications that it offers in environmental biotechnology. Engineered sulfate ...

Protocols for Quantifying Transferable Pesticide Residues in Turfgrass Systems

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes onto humans. For this reason, transferable pesticide residue experiments are required for registration and re-registration by the United States Environmental Protection Agency (USEPA). Although such experiments are required, limited specificity is required pertaining to experimental approach. Experimental approaches used to assess pesticide transfer to humans including hand wiping with cotton gloves, modified California roller (moving a roller of known mass over cotton cloth) and soccer ball roll (ball wrapped with sorbent strip) over three treated turfgrass species (creeping bentgrass, hybrid bermudagrass and tall fescue maintained at 0.4, 5 and 9 cm, respectively) are presented. The modified California roller is the most extensively utilized approach to date, and is best suited for use at low mowing heights due to its reproducibility and large sampling area. The soccer ball roll is a less aggressive transfer approach; however, it mimics a very common occurrence in the most popular international sport, and has many implications for nondietary pesticide exposure from hand-to-mouth contact. Additionally, this approach may be adjusted for other athletic activities with limited modification. Hand wiping is the best approach to transfer pesticides at higher mowing heights, as roller-based approaches can lay blades over; however, it is more subjective due to more variable sampling pressure. Utility of these methods across turfgrass species is presented, and additional considerations to conduct transferable pesticide residue research in turfgrass systems are discussed.

Transferable pesticide residue experiments in turfgrass systems are integral components of human risk exposure assessments. Experimental approaches to measure transferable residues should be adjusted to the human interaction of interest and turfgrass system dynamics. Three transferable pesticide residue protocols are presented and the suitability across three turfgrass systems is discussed.

Plant canopies in established turfgrass systems can intercept an appreciable amount of sprayed pesticides, which can be transferred through various routes ...

Continuous Instream Monitoring of Nutrients and Sediment in Agricultural Watersheds

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in water resources is a prerequisite for understanding the drivers of the pollutant loads and for making informed water resource management decisions. The commonly used &#34;grab sampling&#34; method provides the concentrations of pollutants at the time of sampling (i.e., a snapshot concentration) and may under- or overpredict the pollutant concentrations and loads. Continuous monitoring of nutrients and sediment has recently received more attention due to advances in computing, sensing technology, and storage devices. This protocol demonstrates the use of sensors, sondes, and instrumentation to continuously monitor in situ nitrate, ammonium, turbidity, pH, conductivity, temperature, and dissolved oxygen (DO) and to calculate the loads from two streams (ditches) in two agricultural watersheds. With the proper calibration, maintenance, and operation of sensors and sondes, good water quality data can be obtained by overcoming challenging conditions such as fouling and debris buildup. The method can also be used in watersheds of various sizes and characterized by agricultural, forested, and/or urban land.

With the advancement of technology and the rise in end-user expectations, the need and use of higher temporal resolution data for pollutant load estimation has increased. This protocol describes a method for continuous in situ water quality monitoring to obtain higher temporal resolution data for informed water resource management decisions.

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in ...

Wastewater Irrigation Impacts on Soil Hydraulic Conductivity: Coupled Field Sampling and Laboratory Determination of Saturated Hydraulic Conductivity

Since the early 1960s, an alternative wastewater discharge practice at The Pennsylvania State University has been researched and its impacts monitored. Rather than discharging treated wastewater to a stream, and thereby directly impacting the stream quality, the effluent is applied to forested and cropped land managed by the University. Concerns related to reductions in soil hydraulic conductivity occur when considering wastewater reuse. The methodology described in this manuscript, matching soil sample size with the size of the laboratory-based hydraulic conductivity measurement apparatus, provides the benefits of a relatively rapid collection of samples with the benefits of controlled laboratory boundary conditions. The results suggest that there may have been some impact of wastewater reuse on the soil&#39;s ability to transmit water at deeper depths in the depressional areas of the site. Most of the reductions in the soil hydraulic conductivity in the depressions appear to be related to the depth from which the sample was collected, and by inference, associated with the soil structural and textural differences.

Here we present a methodology which matches a soil sample size and a hydraulic conductivity measurement device to prevent the so-called wall flow along the inside of the soil container from being erroneously included in water flow measurements. Its use is demonstrated with samples collected from a wastewater irrigation site.

Since the early 1960s, an alternative wastewater discharge practice at The Pennsylvania State University has been researched and its impacts monitored. ...

Microalgal Cultivation for Biomethane Obtention from Swine Waste Biogas

Biogas Purification through the use of a Microalgae-Bacterial System in Semi-Industrial High Rate Algal Ponds

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of polluting gases such as carbon dioxide and hydrogen sulfide to increase the content of methane. In this study, we used a microalgal cultivation technology to treat and purify biogas produced from organic waste from the swine industry to obtain ready-to-use biomethane. For cultivation and purification, two 22.2 m3 open-pond photobioreactors coupled with an absorption-desorption column system were set up in San Juan de los Lagos, Mexico. Several recirculation liquid/biogas ratios (L/G) were tested to obtain the highest removal efficiencies; other parameters, such as pH, dissolved oxygen (DO), temperature, and biomass growth, were measured. The most efficient L/Gs were 1.6 and 2.5, resulting in a treated biogas effluent with a composition of 6.8%vol and 6.6%vol in CO2, respectively, and removal efficiencies for H2S up to 98.9%, as well as maintaining O2 contamination values of less than 2%vol. We found that pH greatly determines CO2 removal, more so than L/G, during cultivation because of its participation in the photosynthetic process of microalgae and its ability to vary pH when solubilized due to its acidic nature. DO, and temperature oscillated as expected from the light-dark natural cycles of photosynthesis and the time of day, respectively. Biomass growth varied with CO2 and nutrient feeding as well as reactor harvesting; however, the trend remained primed for growth.

Author Spotlight: Scaling Microalgal Biotechnology for Enhanced Biomethane Production 

Air pollution impacts the quality of life of all organisms. Here, we describe the use of microalgae biotechnology for the treatment of biogas (simultaneous removal of carbon dioxide and hydrogen sulfide) and the production of biomethane through semi-industrial open high-rate algal ponds and subsequent analysis of treatment efficiency, pH, dissolved oxygen, and microalgae growth.

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of ...

A Novel Bioreactor for High Density Cultivation of Diverse Microbial Communities

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Past studies have evaluated the performance of the reactor for the removal of COD1 and nitrogen species2-4 by heterotrophic and chemoautotrophic bacteria, respectively. The HDBR design eliminates the requirement for external flocculation/sedimentation processes while still yielding effluent containing low suspended solids. In this study, the HDBR is applied as a photobioreactor (PBR) in order to characterize the nitrogen removal characteristics of an algae-based photosynthetic microbial community. As previously reported for this HDBR design, a stable biomass zone was established with a clear delineation between the biologically active portion of the reactor and the recycling reactor fluid, which resulted in a low suspended solid effluent. The algal community in the HDBR was observed to remove 18.4% of total nitrogen species in the influent. Varying NH4+ and NO3- concentrations in the feed did not have an effect on NH4+ removal (n=44, p=0.993 and n=44, p=0.610 respectively) while NH4+ feed concentration was found to be negatively related with NO3- removal (n=44, p=0.000) and NO3- feed concentration was found to be positively correlated with NO3- removal (n=44, p=0.000). Consistent removal of NH4+, combined with the accumulation of oxidized nitrogen species at high NH4+ fluxes indicates the presence of ammonia- and nitrite-oxidizing bacteria within the microbial community.

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Here, the HDBR is successfully applied in a photobioreactor (PBR) configuration for the study of nitrogen metabolism by a mixed high density algal community.

A novel reactor design, coined a high density bioreactor (HDBR), is presented for the cultivation and study of high density microbial communities. Past ...

Bioengineering

Spatial Multiobjective Optimization of Agricultural Conservation Practices using a SWAT Model and an Evolutionary Algorithm

Finding the cost-efficient (i.e., lowest-cost) ways of targeting conservation practice investments for the achievement of specific water quality goals across the landscape is of primary importance in watershed management. Traditional economics methods of finding the lowest-cost solution in the watershed context (e.g.,5,12,20) assume that off-site impacts can be accurately described as a proportion of on-site pollution generated. Such approaches are unlikely to be representative of the actual pollution process in a watershed, where the impacts of polluting sources are often determined by complex biophysical processes. The use of modern physically-based, spatially distributed hydrologic simulation models allows for a greater degree of realism in terms of process representation but requires a development of a simulation-optimization framework where the model becomes an integral part of optimization.
Evolutionary algorithms appear to be a particularly useful optimization tool, able to deal with the combinatorial nature of a watershed simulation-optimization problem and allowing the use of the full water quality model. Evolutionary algorithms treat a particular spatial allocation of conservation practices in a watershed as a candidate solution and utilize sets (populations) of candidate solutions iteratively applying stochastic operators of selection, recombination, and mutation to find improvements with respect to the optimization objectives. The optimization objectives in this case are to minimize nonpoint-source pollution in the watershed, simultaneously minimizing the cost of conservation practices. A recent and expanding set of research is attempting to use similar methods and integrates water quality models with broadly defined evolutionary optimization methods3,4,9,10,13-15,17-19,22,23,25. In this application, we demonstrate a program which follows Rabotyagov et al.'s approach and integrates a modern and commonly used SWAT water quality model7 with a multiobjective evolutionary algorithm SPEA226, and user-specified set of conservation practices and their costs to search for the complete tradeoff frontiers between costs of conservation practices and user-specified water quality objectives. The frontiers quantify the tradeoffs faced by the watershed managers by presenting the full range of costs associated with various water quality improvement goals. The program allows for a selection of watershed configurations achieving specified water quality improvement goals and a production of maps of optimized placement of conservation practices.

This work demonstrates an integration of a water quality model with an optimization component utilizing evolutionary algorithms to solve for optimal (lowest-cost) placement of agricultural conservation practices for a specified set of water quality improvement objectives. The solutions are generated using a multi-objective approach, allowing for explicit quantification of tradeoffs.

Finding the cost-efficient (i.e., lowest-cost) ways of targeting conservation practice investments for the achievement of specific water quality goals ...