Name: a flow-through exposure system for evaluating suspended sediments effects on aquatic life
Uploaded: 2017-01-09T13:00:00
Description: Watch this Scientific Journal Video about A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life at JoVE.com

This research was funded by the U.S. Army Corps of Engineers Dredging Operations and Environmental Research Program, Todd Bridges, Director. Permission was granted by the Chief of Engineers to publish this material.

All FLEES experiments with vertebrates were performed under the appropriate Engineer Research and Development Center (ERDC) Environmental Laboratory Institutional Animal Care and Use Protocols.
1. FLEES Modules, Water Bath, and Aquaria
<ol>
	<li>Obtain wood posts, studs and plywood for building the module. Construct the modules (in number and size) similar to a basic workbench to meet research objectives.
	<ol>
		<li>Cut the plywood (0.127 cm) for the top and shelf. Cut the posts (10.16 x 10...

<ol>
	<li>National Research Council (NRC). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+process+for+setting,+managing,+and+monitoring+environmental+windows+for+dredging+projects.">A process for setting, managing, and monitoring environmental windows for dredging projects.</a> Marine Board, Transportation Research Board, Special Report 262. , (2001).</li><li>Suedel, B. C., Kim, J., Clarke, D. G., Linkov, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+risk-informed+decision+framework+for+setting+environmental+windows+for+dredging+projects.">A risk-informed decision framework for setting environmental windows for dredging projects.</a> Sci Total Environ. 403, 1-11 (2008).</li><li>Reine, K. J., Dickerson, D. D., Clarke, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+windows+associated+with+dredging+operations.">Environmental windows associated with dredging operations.</a> DOER Technical Notes Collection. ERDC TN DOER-E2. , (1998).</li><li>Wilber, D. H., Clarke, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+effects+of+suspended+sediments:+A+review+of+suspended+sediment+impacts+on+fish+and+shellfish+with+relation+to+dredging+activities+in+estuaries.">Biological effects of suspended sediments: A review of suspended sediment impacts on fish and shellfish with relation to dredging activities in estuaries.</a> N Am J Fish Manag. 21, 855-875 (2001).</li><li>Suedel, B. C., Lutz, C. H., Clarke, J. U., Clarke, D. G. <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+suspended+sediment+on+walleye+(Sander+vitreus)+eggs.">The effects of suspended sediment on walleye (Sander vitreus) eggs.</a> J. Soils Sediments. 12, 995-1003 (2012).</li><li>Travis, J., Kring, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> LabVIEW for Everyone: Graphical Programming Made Easy and Fun (National Instruments Virtual Instrumentation Series). , (2006).</li><li>Hazardous Waste Test Methods/SW-846 On-line. Office of Solid Waste and Emergency Response (OSWER) Available from: <a target="_blank" href="https://www.epa.gov/hw-sw846/sw-846-compendium">https://www.epa.gov/hw-sw846/sw-846-compendium</a> (2016)</li><li>Plumb, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Procedure for handling and chemical analysis of sediment and water samples, EPA/CE-81-1. , (1981).</li><li>Clarke, D. G., Wilber, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Assessment of potential impacts of dredging operations due to sediment resuspension. , (2000).</li><li>Suedel, B. C., Clarke, J. U., Lutz, C. H., Clarke, D. G., Godard-Codding, C., Maul, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suspended+sediment+effects+on+walleye+(Sander+vitreus).">Suspended sediment effects on walleye (Sander vitreus).</a> J. Great Lakes Res. 40, 141-148 (2014).</li><li>Wilkens, J. L., Katzenmeyer, A. W., Hahn, N. M., Hoover, J. J., Suedel, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+test+of+suspended+sediment+effects+on+short-term+survival+and+swimming+performance+of+juvenile+Atlantic+Sturgeon+(Acipenser+oxyrinchus+oxyrinchus).">Laboratory test of suspended sediment effects on short-term survival and swimming performance of juvenile Atlantic Sturgeon (Acipenser oxyrinchus oxyrinchus).</a> J. Appl. Ichthy. 31, 984-990 (2015).</li><li>Suedel, B. C., Clarke, J. U., Wilkens, J., Lutz, C. H., Clarke, D. G. <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+a+simulated+sediment+plume+on+eastern+oyster+(Crassostrea+virginica)+survival,+growth,+and+condition.">The effects of a simulated sediment plume on eastern oyster (Crassostrea virginica) survival, growth, and condition.</a> Estuaries and Coasts. 38 (2), 578-589 (2015).</li><li>Bilotta, G. S., Brazier, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+influence+of+suspended+solids+on+water+chemistry+and+aquatic+biota.">Understanding the influence of suspended solids on water chemistry and aquatic biota.</a> Water Res. 42, 2849-2861 (2008).</li><li>Reine, K., Clarke, D., Dickerson, C., Pickard, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+potential+impacts+of+bucket+dredging+plumes+on+walleye+spawning+habitat+in+Maumee+Bay,+Ohio.">Assessment of potential impacts of bucket dredging plumes on walleye spawning habitat in Maumee Bay, Ohio.</a> Proceedings of the 18th World Dredging Congress (WODCON XVIII). , (2007).</li></ol>

The FLEES technology improves on existing methods4,9 by maintaining and controlling suspended sediments over a wide range of exposure times and suspended sediment concentrations using an automated, computer controlled system. The technology is flexible such that it can be used to evaluate the effects of suspended sediments to multiple aquatic species and life stages of varying sizes from eggs to adults depending on the species. In the future, the technology is capable of assessing suspended sediment effects to...

Dredging operations use mechanical methods to remove bottom sediments from harbors and navigation channels. During removal, some portion of the disturbed sediment is suspended into the water column, potentially making this a source of physical stress to aquatic species. In addition to being suspended, the sediment may be transported away from the dredge by ambient conditions before settling out of the water column. The combination of these two mechanisms means that an aquatic organism occurring near an operating dredge may be exposed to suspended sediments and suffer adverse effects. To address such concerns, environmental windows (seasonal dredging restrictions) are routinely used as a management practice to reduce or eliminate risk of potentially harmful impacts of suspended sediments from dredging activities on aquatic resources1,2.
Environmental windows are most commonly established to protect endangered, threatened or commercially valuable species such as the walleye (Sander vitreus) and eastern oyster (Crassostrea virginica)3. The supporting justification for imposing environmental windows often focuses on how dredge activities may potentially physically disturb (e.g., suspended sediment) an animal&#39;s ability to complete a specific part of its life history. The life stages commonly cited are eggs and larvae for keeping migration routes open for anadromous species3. However, there is limited information concerning species-specific biological effects relevant to suspended sediment4,5 available to inform using environmental windows as a risk management tool.
For these reasons, the FLEES was designed, built, and used to simulate the suspension of sediment, and to determine its effects on early life stages of aquatic organisms. FLEES studies use fine-grained sediment particles (i.e., predominantly silts, clays, and fine sands) which are most likely to remain in suspension and migrate furthest from the source. The FLEES is capable of testing fish eggs and larvae, but it can also be retrofitted to accommodate other aquatic organisms, making it a unique capability. The resulting biological response data can then be used to assess the effects of suspended sediments. The following procedures provide an overview of how the technology can be constructed and operated to yield repeatable suspended sediment concentrations and effects data using various aquatic species.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Parts List for one FLEES Module, Water Bath, and Aquarium</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>post, wood - used to build module (cut to 78 in)</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 4 
			Size: 4 in x 4 in x 8 ft</td>
		</tr>
		<tr>
			<td>plywood, marine grade - fastened to wooden posts about 18 in off ground - for holding water bath (60 in x 42 in)</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 1 
			Size: 3/4 in x 4 ft x 8 ft</td>
		</tr>
		<tr>
			<td>plywood - fastened on top of wooden posts - for holding pipes, solenoids and electrical (60 in x 42 in)</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 1 
			Size: 1/4 in x 4 ft x 8 ft</td>
		</tr>
		<tr>
			<td>stud, wood - used to brace plywood and wooden posts (cut to fit)</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 4 
			Size: 2 in x 4 in x 96 in</td>
		</tr>
		<tr>
			<td>tank, fiberglass - water bath with two drains: 1) to supply chiller; and 2) to drain water</td>
			<td>Hydro Composites, LLC, Stockdale, TX, USA</td>
			<td>FBT-226</td>
			<td>Quantity: 1 
			Size: 150-gal</td>
		</tr>
		<tr>
			<td>chiller, water with self contained pump - for water bath; chiller sits under module</td>
			<td>Remcor Products Co., Glendale Heights, IL, USA</td>
			<td>CFF-500</td>
			<td>Quantity: 1 
			Size: 1/2 hp</td>
		</tr>
		<tr>
			<td>tank, domed bottom - FLEES aquaria - sit inside water bath</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>5197</td>
			<td>Quantity: 5 
			Size: 19 L</td>
		</tr>
		<tr>
			<td>tank, stand - acrylic stand, 12 in x 12 in x 6 in - to hold aquaria</td>
			<td>custom built by ERDC shops</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: custom</td>
		</tr>
		<tr>
			<td>urethane, liquid- to seal carboy plug and prevent leaking</td>
			<td>Forsch Polymer Corporation, Englewood, CO</td>
			<td>60A Liquid Urethane</td>
			<td>Quantity: 5 
			Size: epoxy kit</td>
		</tr>
		<tr>
			<td>pump, magnetic drive - to suspend sediment in each aquarium</td>
			<td>March Manufacturing Inc., Glenview, IL, USA</td>
			<td>MDX-3-1/2 115 v</td>
			<td>Quantity: 5 
			Size: 28 liter per min</td>
		</tr>
		<tr>
			<td>light, LED - installed over water bath</td>
			<td>C2 Development, Inc., Ames, IA, USA</td>
			<td>Hydra 26</td>
			<td>Quantity: 2 
			Size: based on area to light</td>
		</tr>
		<tr>
			<td>pipe, PVC schedule 40 - installed in drain of water bath to control water level</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: - 
			Size: 1 in</td>
		</tr>
		<tr>
			<td>fittings, bulkhead - for aquaria/water bath connections to pumps, drains, water and slurry lines</td>
			<td>Lifegard Aquatics, Cerritos, CA, USA</td>
			<td>R270900</td>
			<td>Quantity: 20 
			Size: 1/2 in FPT x FPT</td>
		</tr>
		<tr>
			<td>fittings, quick-disconnect, male pipe threaded inserts - insert in tank bulkhead</td>
			<td>Cole-Parmer, Vernon Hills, IL, USA</td>
			<td>EW-31303-36</td>
			<td>Quantity: 10 
			Size: 1/2 in MPT</td>
		</tr>
		<tr>
			<td>fittings, quick-disconnect, valved hose barbs - connection between aquarium and insert in tank bulkhead</td>
			<td>Cole-Parmer, Vernon Hills, IL, USA</td>
			<td>EW-31303-11</td>
			<td>Quantity: 10 
			Size: 1/2 in</td>
		</tr>
		<tr>
			<td>fittings, black HDPE threaded elbow - for aquaria vinyl tube connections to slurry/water line and pump</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>62043</td>
			<td>Quantity: 20 
			Size: 1/2 NPT x 1/2 in Hose ID</td>
		</tr>
		<tr>
			<td>fittings, black HDPE threaded adapter - for connections between pump and tank bulkhead</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>62017</td>
			<td>Quantity: 10 
			Size: 1/2 NPT x 1/2 in Hose ID</td>
		</tr>
		<tr>
			<td>tube, vinyl - connect slurry/water line to aquaria and to connect pumps to aquaria</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 25 ft 
			Size: 1/2 in ID</td>
		</tr>
		<tr>
			<td>tube, vinyl - connect to aquaria drains inserts and water bath drain</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 25 ft 
			Size: 5/8 in ID</td>
		</tr>
		<tr>
			<td>clamp, hose, stainless steel - to clamp vinyl tube to hose barbs</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 40 
			Size: #8</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments 
			&#160;</td>
		</tr>
		<tr>
			<td>Parts list for Slurry System</td>
			<td></td>
			<td></td>
			<td> 
			&#160;</td>
		</tr>
		<tr>
			<td>chiller, water with self contained pump - sits off to side of slurry tank</td>
			<td>Remcor Products Co., Glendale Heights, IL, USA</td>
			<td>CFF-500</td>
			<td>Quantity: 1 
			Size: 1/2 hp</td>
		</tr>
		<tr>
			<td>125 gallon open top cone bottom tank w/Stand - 42 in x 35 in - contains the water and sediment to make slurry</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>8586</td>
			<td>Quantity: 1 
			Size: 125 gal</td>
		</tr>
		<tr>
			<td>Cover for 125 gallon tank</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>8935</td>
			<td>Quantity: 1 
			Size: 42 in x 35 in</td>
		</tr>
		<tr>
			<td>valve, PVC - connect tank drain to pump - isolate for maintenance</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 2 
			Size: 1-1/2 in</td>
		</tr>
		<tr>
			<td>pump, double diaphragm mounted on stand - used to recirculate slurry</td>
			<td>Wilden-pumps.co.uk &#38; Air Pumping Ltd., Essex, UK</td>
			<td>P2/PPPP/WF/WF/PTV/400</td>
			<td>Quantity: 1 
			 
			contact distributor</td>
		</tr>
		<tr>
			<td>sensor, optical backscatter - measure NTU in slurry tank</td>
			<td>Campbell Scientific, Logan, UT, USA</td>
			<td>OBS-3+</td>
			<td>Quantity: 1 
			Size: 0-1000 NTU</td>
		</tr>
		<tr>
			<td>pipe, PVC Schedule 40 - to recirculate slurry</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 20 ft 
			Size: 1 in</td>
		</tr>
		<tr>
			<td>pipe, flexible PVC - fitted with union and used to connect to next module</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 10 ft 
			Size: 1 in</td>
		</tr>
		<tr>
			<td>union, PVC Schedle 80 Socket - connect slurry line with next module</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 8 
			Size: 1/2 in</td>
		</tr>
		<tr>
			<td>solenoid, plastomatic (normally closed) - introduce slurry</td>
			<td>Plast-O-Matic Valves, Inc., Cedar Grove, NJ, USA</td>
			<td>EASYMT4V12R24-PV</td>
			<td>Quantity: 5 
			Size: 1/2 in NPT threaded, 24 VAC 
			contact distributor</td>
		</tr>
		<tr>
			<td>fitting, PVC tee - connect slurry pipe with solenoid</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 1 in x 1 in x 1 in slip x slip x FIPT</td>
		</tr>
		<tr>
			<td>fittings, 1 in PVC ball valve threaded - shut off for slurry delivery to solenoid/water lines</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 7 
			Size: 1/2 in</td>
		</tr>
		<tr>
			<td>fittings, 1 in PVC union threaded - connect slurry solenoid to shut off valve</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 1/2 in</td>
		</tr>
		<tr>
			<td>tube, vinyl - connection between water solenoid and slurry solenoid</td>
			<td>Local vendor</td>
			<td>N/A</td>
			<td>Quantity: 50 ft 
			Size: 1/4&#34; ID</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments 
			&#160;</td>
		</tr>
		<tr>
			<td>Parts list for Water System</td>
			<td></td>
			<td></td>
			<td> 
			&#160;</td>
		</tr>
		<tr>
			<td>chiller, water with self contained pump - sits off to side of reservoir</td>
			<td>Remcor Products Co., Glendale Heights, IL, USA</td>
			<td>CFF-500</td>
			<td>Quantity: 1 
			Size: 1/2 hp</td>
		</tr>
		<tr>
			<td>125 gallon open top cone bottom tank w/Stand - 42 in x 35 in - contains the water and sediment to make slurry</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>8586</td>
			<td>Quantity: 1 
			Size: 125 gal</td>
		</tr>
		<tr>
			<td>Cover for 125 gallon tank</td>
			<td>United States Plastic Corp, Lima, OH, USA</td>
			<td>8935</td>
			<td>Quantity: 1 
			Size: 42 in x 35 in</td>
		</tr>
		<tr>
			<td>valve, PVC - connect tank drain to water pump</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 2 
			Size: 1 in</td>
		</tr>
		<tr>
			<td>pump, magnetic drive, in-line use - used to recirculate water to aquaria and chiller</td>
			<td>Little Giant, Fort Wayne, IN, USA</td>
			<td>3-MD-SC</td>
			<td>Quantity: 1 
			Size: 1/12 hp</td>
		</tr>
		<tr>
			<td>solenoid, alco - introduce water</td>
			<td>discontinued; ASCO, Florham Park, NJ,USA for similar</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 24 v, 1/4 in NIPT</td>
		</tr>
		<tr>
			<td>fittings, black HDPE reducer connector - connect 1/4 in hose water line from solenoid&#160; to 1/2 in hose</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 1/2 in hose ID x 1/4 in hose ID</td>
		</tr>
		<tr>
			<td>fittings, black HDPE tee - connect 1/2 in hose water line and slurry to aquaria</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 1/2 in NPT x 1/2 in hose ID x 1/2 in hose ID</td>
		</tr>
		<tr>
			<td>fittings, street elbow</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 1/2 in 90&#176; MIPT x FIPT</td>
		</tr>
		<tr>
			<td>fittings, PVC threaded pipe nipples - connect union fittings with solenoids and other connections</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 12 
			Size: 1/2 in</td>
		</tr>
		<tr>
			<td>fittings, union threaded - connect slurry/water lines with next module</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 6 
			Size: 1 in PVC&#160;</td>
		</tr>
		<tr>
			<td>fittings, reducer bushing - connect to reducer tee in water line</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size: 1/2 in male by 1/4 in female FIPT</td>
		</tr>
		<tr>
			<td>fittings, threaded pipe nipples - connection between bushing and water solenoid</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 
			Size:&#160; 4 in long&#160; x 1/4 in&#160;</td>
		</tr>
		<tr>
			<td>pipe, PVC&#160; - make connections between tank, pump and chiller</td>
			<td>local plumbing vendor</td>
			<td>N/A</td>
			<td>Quantity: 5 ft 
			Size: Schedule 40</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments 
			&#160;</td>
		</tr>
		<tr>
			<td>Parts list for Sensors, Data Acquisition Device, and Computer Software</td>
			<td></td>
			<td></td>
			<td> 
			&#160;</td>
		</tr>
		<tr>
			<td>software, LabView</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>LabView 2015 Base</td>
			<td>Quantity: 1 
			Size: N/A</td>
		</tr>
		<tr>
			<td>SCXI-1001 12-Slot Chassis, U.S. 120 VAC</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>776571-01</td>
			<td>Quantity: 1 
			Size: N/A</td>
		</tr>
		<tr>
			<td>SCXI 1100 - 32-Channel, &#177;10 V Analog Input Module</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>776572-00</td>
			<td>Quantity: 1 
			Size: N/A</td>
		</tr>
		<tr>
			<td>SCXI 1303 - Terminal block designed for high-accuracy thermocouple measurements</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>777687-03</td>
			<td>Quantity: 2 
			Size: N/A</td>
		</tr>
		<tr>
			<td>SCXI 1102B - 32-Channel Thermocouple/Voltage Input Module</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>776572-02B</td>
			<td>Quantity: 1 
			Size: N/A</td>
		</tr>
		<tr>
			<td>SCXI 1161 - General-Purpose Relay Module</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>776572-61</td>
			<td>Quantity: 6 
			Size: N/A</td>
		</tr>
		<tr>
			<td>SCXI 1300 - General-Purpose Voltage Module</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>777687-00</td>
			<td>Quantity: 1 
			Size: N/A</td>
		</tr>
		<tr>
			<td>PCMCIA Card DAQCARD-AI-16E-4</td>
			<td>National Instruments, Austin, Texas, USA</td>
			<td>N/A - legacy</td>
			<td>Quantity: 1 
			Size: N/A 
			used cards available online</td>
		</tr>
		<tr>
			<td>sensor, optical backscatter - measure NTU in each aquarium</td>
			<td>Campbell Scientific Inc., Logan, UT, USA</td>
			<td>OBS-3+</td>
			<td>Quantity: 5 
			Size: 0-1000 NTU</td>
		</tr>
	</tbody>
</table>

a flow-through exposure system for evaluating suspended sediments effects on aquatic life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A series of operational runs is performed before beginning an experiment to ensure that the FLEES is delivering the appropriate concentrations of sediments to each aquarium (Sections 5.5 and 6.2). Figure 6 illustrates how NTU concentrations are maintained in experimental aquaria to achieve target suspended sediment concentrations. In this example, FLEES evaluated whether suspended sediment could be maintained over a three-day period with the proposed test sediment, a dura...

Watch this Scientific Journal Video about A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life at JoVE.com

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

The overall goal of this experimental technology is delivering a robust and flexible flow-through exposure system for maintaining sediment and suspension, to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory. This technology can help us answer key questions in the field of environmental toxicology to improve our understanding about the effects of physical stressors such as suspended sediments in aquatic environments. The main advantage of this technology is the ability to simulate sediment suspension for incoming management practices to reduce the interactions between aquatic organisms and suspended sediments.Though this method can provide insight into the effects of dredging, it can also be used to evaluate the effects of suspended sediments resulting from vessel traffic, freshets, and storms. Visual demonstration of this technology is critical, as steps in its construction combined with its calibration are critical to successful application. Begin building and assembly of the module and water bath as described in the text protocol.To construct an overflow drain, use a hole saw to cut a 2.54 centimeter diameter hole five centimeters from the top of a 19 liter domed bottom polyethylene tank. Install a bulkhead fitting and an insert on the exterior of the bulkhead to serve as the overflow drain. To construct the slurry water inlet, use a hole saw to cut another 2.54 centimeter diameter hole five centimeters from the top of the aquarium.Install another bulkhead fitting and a threaded elbow hose barb. For constructing the pump outlet, cut the same type of hole through the middle of the tank bottom and install a bulkhead fitting. Thread the exterior side of the bulkhead with an elbow hose fitting.To construct the pump inlet, cut another hole located off-center of the tank bottom and install a bulkhead fitting. Thread the exterior side of the bulkhead with an elbow hose fitting. On the exterior side of the water bath tank, measure nine centimeters from the bottom and draw a line along the length of the tank.Following the line, cut a pair of 2.54 centimeter diameter holes along the length of the water bath for each aquarium and install bulkhead fittings. Mount magnetic drive pumps to a stand that will fit under the water bath along the side containing the holes and bulkhead fittings for connecting to the aquaria. Install an inline cord switch for each pump, or wire the pumps to a switch box for power.Thread the exterior side of the water bath tank bulkheads with hose barbs. Attach vinyl tubing to the pump inlet and outlet before connection it to the bulkheads going to the appropriate aquarium. Inside the water bath, install a quick disconnect insert into the bulkhead.After connecting each aquarium to a pump, attach vinyl tubing to the hose barbs installed on the bottom of the aquaria and attach the tubing to the quick disconnect valve hose barb. Next, connect the quick disconnects between the pump and the aquarium. Install a ball valve in this connection to isolate the pump for maintenance purposes.Next, connect each aquarium's overflow drain to common drain via vinyl tubing. Then connect the common drain to the water bath drain. Connect each aquarium slurry water inlet to the slurry and water system installed on top of the module.Connect the slurry tank drain to the inlet of an air-operated double-diaphragm pump that has been mounted onto a stand next to a slurry tank. Incorporate a PVCT to direct the slurry to the pump or to the laboratory drain. Also incorporate valves connecting the tank and the pump to isolate them for maintenance.To power the pump, connect it to the laboratory building's air compressor. Mount a PVC pipe on top of the modules and create a recirculation line to provide the slurry to each aquarium upon demand. At the point of use located farthest from the slurry reservoir, install a return line to transport the unused slurry back to the reservoir.Use flexible PVC and union fittings to connect between the modules. Connect the slurry solenoid valves to the recirculating slurry pipe using tees, ball valves, and union fittings, to isolate solenoids from the main PVC pipe for maintenance. Insuring that each solenoid valve is located above the aquarium that it supplies, connect the solenoid valves to the corresponding aquaria as described in the text protocol.Install an optical backscatter sensor or OBS in each aquarium next to the slurry water inlet to measure turbidity. Position the sensor so that it is submerged about five centimeters below the water surface with the sensor facing towards the middle of the tank. Use a clamp or other device to mount the sensor.Install an OBS in the slurry reservoir and position the sensor so that it is completely submerged about 20 centimeters below the water surface. Next, wire the water and slurry solenoid valves. The OBS is located in each aquarium and slurry tank and a thermal couple located at each water bath, into electrical junction boxes mounted on top of the module into a data acquisition device.Install quick disconnects at the terminal ends of all wiring wherever possible. Proceed to design a computer application for data acquisition, instrument control, and automation as described in the text protocol. To prepare for the experiment start the pump to recirculate carbon filtered tap water that has been used to fill the slurry tank.In a separate container, use a mechanical mixer to homogenize the test sediment. After the sediment is homogenized, remove a small portion and introduce it into the slurry tank using a graduated polypropylene beaker. Continue to introduce sediment until 1000 NTU is achieved.In the program, navigate to the water bath tab and start all aquarium profiles. After operating the FLEES for at least one hour so NTUs can stabilize in each aquarium, turn the data logging on to record NTU readings by each aquarium OBS. For TSS measurement, collect three 100 milliliter water samples from each aquarium assigned a TSS treatment of less than 500 milligrams per liter.Separately collect three 50 milliliter water samples from each aquarium assigned a TSS treatment of greater than or equal to 500 milligrams per liter. Measure the TSS by vacuum filtering the samples through pre-weighed 0.45 micron filter paper. Immediately post-filtering, dry the filter and contents at 105 degrees Celsius for a minimum of four hours.And then reweigh to the nearest 0.1 milligram. Use the average of the three samples as a measure of TSS in each aquarium. To perform the experiment, first turn on all FLEES hardware used for data acquisition, instrument control, and automation.After filling the aquaria, water baths, and water reservoir with the desired test water, start all water chiller heat exchangers. Also, confirm and adjust the light cycle. Start the pump to recirculate carbon filtered tap water that has been used to fill the slurry tank.Then, remove a small portion of homogenize sediment and introduce it into the slurry tank. Continue to introduce sediment until 1000 NTU is reached. Then create a profile for each aquarium as described in the text protocol.After animals are stocked into the aquaria as described in the text protocol access the GUI and navigate to the water bath tab to start all aquarium profiles. Following operation of the FLEES for at least one hour, turn the data logging on to record NTU readings by each aquarium OBS. NTU and corresponding TSS concentrations are maintained in experimental aquaria to achieve target-suspended sediment concentrations.In this example, the FLEES evaluated whether the suspended sediment could be maintained over a three day period. Similarly, NTU and corresponding TSS concentrations can be maintained at considerably lower concentrations representing ambient conditions. The FLEES is calibrated using test sediment to quantify the relationship between TSS and turbidity to ensure target TSS concentrations are achieved.The relationship between NTU and TSS is displayed for an example test sediment evaluated in the FLEES. Once FLEES is constructed and operating properly, it is flexible enough to evaluate the effects of suspended sediments to multiple aquatic species in life stages of varying sizes, from eggs to adults, depending upon the species. It is important to remember that the most critical step in the methodology is to calibrate the FLEES with every test sediment.So the relationship between total suspended solids and turbidity can be quantified, thereby enabling the matching of target TSS concentrations. After its development, the FLEES technology paved the way for researchers in the field of environmental toxicology to investigate the effects of suspended sediments on a wide variety of aquatic organisms. After watching this video, you should have a good understanding of how to construct and utilize the technology in your own laboratory.As the automation and programming features enable total suspended solids concentrations to be easily adjusted to meet experimental needs.

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Research

JoVE Journal

Environment

An Aquatic Microbial Metaproteomics Workflow: From Cells to Tryptic Peptides Suitable for Tandem Mass Spectrometry-based Analysis

Meta-omic technologies such as metagenomics, metatranscriptomics and metaproteomics can aid in the understanding of microbial community structure and metabolism. Although powerful, metagenomics alone can only elucidate functional potential. On the other hand, metaproteomics enables the description of the expressed in situ metabolism and function of a community. Here we describe a protocol for cell lysis, protein and DNA isolation, as well as peptide digestion and extraction from marine microbial cells collected on a cartridge filter unit (such as the Sterivex filter unit) and preserved in an RNA stabilization solution (like RNAlater). In mass spectrometry-based proteomics studies, the identification of peptides and proteins is performed by comparing peptide tandem mass spectra to a database of translated nucleotide sequences. Including the metagenome of a sample in the search database increases the number of peptides and proteins that can be identified from the mass spectra. Hence, in this protocol DNA is isolated from the same filter, which can be used subsequently for metagenomic analysis.

This protocol is for the extraction and concentration of protein and DNA from microbial biomass collected from seawater, followed by the generation of tryptic peptides suitable for tandem mass spectrometry-based proteomic analysis.

Meta-omic technologies such as metagenomics, metatranscriptomics and metaproteomics can aid in the understanding of microbial community structure and ...

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most experiments examine only two or a few levels of CA concentration and a single soil type, but if CA can be varied as a gradient from subambient to superambient concentrations on multiple soils, we can discern whether past ecosystem responses may continue linearly in the future and whether responses may vary across the landscape. The Lysimeter Carbon Dioxide Gradient Facility applies a 250 to 500 &#181;l L-1 CA gradient to Blackland prairie plant communities established on lysimeters containing clay, silty clay, and sandy soils. The gradient is created as photosynthesis by vegetation enclosed in in temperature-controlled chambers progressively depletes carbon dioxide from air flowing directionally through the chambers. Maintaining proper air flow rate, adequate photosynthetic capacity, and temperature control are critical to overcome the main limitations of the system, which are declining photosynthetic rates and increased water stress during summer. The facility is an economical alternative to other techniques of CA enrichment, successfully discerns the shape of ecosystem responses to subambient to superambient CA enrichment, and can be adapted to test for interactions of carbon dioxide with other greenhouse gases such as methane or ozone.

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 &#181;l L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

Continuing increases in atmospheric carbon dioxide concentrations (CA) mandate techniques for examining impacts on terrestrial ecosystems. Most ...

Alternative Method of Removing Otoliths from Sturgeon

Extracting the otoliths (ear bones) from fish that have very thick skulls can be difficult and very time consuming. The common practice of making a transverse vertical incision on the top of the skull with a hand or electrical saw may damage the otolith if not performed correctly. Sturgeons (Acipenseridae) are one family in particular that have a very large and thick skull. A new laboratory method entering the brain cavity from the ventral side of the fish to expose the otoliths was easier than other otolith extraction methods found in the literature. Methods reviewed in the literature are designed for the field and are more efficient at processing large quantities of fish quickly. However, this new technique was designed to be more suited for a laboratory setting when time is not pressing and successful extraction from each specimen is critical. The success of finding and removing otoliths using this technique is very high and does not compromise the structure in any manner. This alternative technique is applicable to other similar fish species for extracting the otoliths.

The goal of the protocol is to show an effective method to extract the otoliths from sturgeon carcasses.

Extracting the otoliths (ear bones) from fish that have very thick skulls can be difficult and very time consuming. The common practice of making a ...

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and rates of mortality in populations, which has a variety of applications in ecology, including agricultural ecosystems. Horizontal, or cohort-based, life tables provide for the most direct and accurate method of quantifying vital population rates because they follow a group of individuals in a population from birth to death. Here, protocols are presented for conducting and analyzing cohort-based life tables in the field that takes advantage of the sessile nature of the immature life stages of a global insect pest, Bemisia tabaci. Individual insects are located on the underside of cotton leaves and are marked by drawing a small circle around the insect with a non-toxic pen. This insect can then be observed repeatedly over time with the aid of hand lenses to measure development from one stage to the next and to identify stage-specific causes of death associated with natural and introduced mortality forces. Analyses explain how to correctly measure multiple mortality forces that act contemporaneously within each stage and how to use such data to provide meaningful population dynamic metrics. The method does not directly account for adult survival and reproduction, which limits inference to dynamics of immature stages. An example is presented that focused on measuring the impact of bottom-up (plant quality) and top-down (natural enemies) effects on the mortality dynamics of B. tabaci in the cotton system.

Methodology for Developing Life Tables for Sessile Insects in the Field Using the Whitefly, Bemisia tabaci, in Cotton As a Model System

Life tables allow quantification of the sources and rates of mortality in insect populations and contribute to understanding, predicting and manipulating population dynamics in agroecosystems. Methods for conducting and analyzing cohort-based life tables in the field for an insect with sessile immature life stages are presented.

Life tables provide a means of measuring the schedules of birth and death from populations over time. They also can be used to quantify the sources and ...

Collection and Extraction of Occupational Air Samples for Analysis of Fungal DNA

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several limitations that have resulted in the exclusion of many species. Advances in the field over the last two decades have led occupational health researchers to turn to molecular-based approaches for identifying fungal hazards. These methods have resulted in the detection of many species within indoor and occupational environments that have not been detected using traditional methods. This protocol details an approach for determining fungal diversity within air samples through genomic DNA extraction, amplification, sequencing, and taxonomic identification of fungal internal transcribed spacer (ITS) regions. ITS sequencing results in the detection of many fungal species that are either not detected or difficult to identify to species level using culture or microscopy. While these methods do not provide quantitative measures of fungal burden, they offer a new approach to hazard identification and can be used to determine overall species richness and diversity within an occupational environment.

Determining the fungal diversity within an environment is a method utilized in occupational health studies to identify health hazards. This protocol describes DNA extraction from occupational air samples for amplification and sequencing of fungal ITS regions. This approach detects many fungal species that can be overlooked by traditional assessment methods.

Traditional methods of identifying fungal exposures in occupational environments, such as culture and microscopy-based approaches, have several ...

Experimental Methods of Dust Charging and Mobilization on Surfaces with Exposure to Ultraviolet Radiation or Plasmas

Electrostatic dust transport has been hypothesized to explain a number of observations of unusual planetary phenomena. Here, it is demonstrated using three recently developed experiments in which dust particles are exposed to thermal plasma with beam electrons, beam electrons only, or ultraviolet (UV) radiation only. The UV light source has a narrow bandwidth in wavelength centered at 172 nm. The beam electrons with the energy of 120 eV are created with a negatively biased hot filament. When the vacuum chamber is filled with the argon gas, a thermal plasma is created in addition to the electron beam. Insulating dust particles of a few tens of microns in diameter are used in the experiments. Dust particles are recorded to be lofted to a height up to a few centimeters with a launch speed up to 1 m/s. These experiments demonstrate that photo and/or secondary electron emission from a dusty surface changes the charging mechanism of dust particles. According to the recently developed &#34;patched charge model&#34;, the emitted electrons can be re-absorbed inside microcavities between neighboring dust particles below the surface, causing the accumulation of enhanced negative charges on the surrounding dust particles. The repulsive forces between these negatively charged particles may be large enough to mobilize and lift them off the surface. These experiments present the advanced understanding of dust charging and transport on dusty surfaces, and laid a foundation for future investigations of its role in the surface evolution of airless planetary bodies.

Dust charging and mobilization is demonstrated in three experiments with exposure to thermal plasma with beam electrons, beam electrons only, or ultraviolet (UV) radiation only. These experiments present the advanced understanding of electrostatic dust transport and its role in shaping the surfaces of airless planetary bodies.

Electrostatic dust transport has been hypothesized to explain a number of observations of unusual planetary phenomena. Here, it is demonstrated using ...

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol for performing in situ juvenile exposure tests within oligotrophic river catchments over one-month and three-month periods. Two methods (in both modifications) are presented to evaluate the juvenile growth and survival rate. The methods and modifications differ in value for the locality bioindication and each has its benefits as well as limitations. The sandy cage method works with a large set of individuals, but only some of the individuals are measured and the results are evaluated in bulk. In the mesh cage method, the individuals are kept and measured separately, but a low individual number is evaluated. The open water exposure modification is relatively easy to apply; it shows the juvenile growth potential of sites and can also be effective for water toxicity testing. The within-bed exposure modification needs a high workload but is closer to the conditions of a natural juvenile environment and it is better for reporting the real suitability of localities. On the other hand, more replications are needed in this modification due to its high-hyporheic environment variability.

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

In situ bioindications enable determination of the suitability of an environment for endangered mussel species. We describe two methods based on the juvenile exposure of freshwater pearl mussels in cages to oligotrophic river habitats. Both methods are implemented in variants for open water and hyporheic water environments.

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol ...

Characterization of Aquatic Biofilms with Flow Cytometry

Biofilms are dynamic consortia of microorganism that play a key role in freshwater ecosystems. By changing their community structure, biofilms respond quickly to environmental changes and can be thus used as indicators of water quality. Currently, biofilm assessment is mostly based on integrative and functional endpoints, such as photosynthetic or respiratory activity, which do not provide information on the biofilm community structure. Flow cytometry and computational visualization offer an alternative, sensitive, and easy-to-use method for assessment of the community composition, particularly of the photoautotrophic part of freshwater biofilms. It requires only basic sample preparation, after which the entire sample is run through the flow cytometer. The single-cell optical and fluorescent information is used for computational visualization and biological interpretation. Its main advantages over other methods are the speed of analysis and the high-information-content nature. Flow cytometry provides information on several cellular and biofilm traits in a single measurement: particle size, density, pigment content, abiotic content in the biofilm, and coarse taxonomic information. However, it does not provide information on biofilm composition on the species level. We see high potential in the use of the method for environmental monitoring of aquatic ecosystems and as an initial biofilm evaluation step that informs downstream detailed investigations by complementary and more detailed methods.

Flow cytometry in combination with visual clustering offers an easy-to-use and fast method for studying aquatic biofilms. It can be used for biofilm characterization, detection of changes in biofilm community structure, and detection of abiotic particles embedded in the biofilm.

Biofilms are dynamic consortia of microorganism that play a key role in freshwater ecosystems. By changing their community structure, biofilms respond ...

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and toxicological studies. Using automated digital tracking platforms, recent efforts in neuropharmacology are leveraging larval fish locomotor behaviors to identify potential therapeutic targets for novel small molecules. Similar to these efforts, research in the environmental sciences and comparative pharmacology and toxicology is examining various behaviors of fish models as diagnostic tools in tiered evaluation of contaminants and real-time monitoring of surface waters for contaminant threats. Whereas the zebrafish is a popular larval fish model in the biomedical sciences, the fathead minnow is a common larval fish model in ecotoxicology. Unfortunately, fathead minnow larvae have received considerably less attention in behavioral studies. Here, we develop and demonstrate a behavioral profile protocol using caffeine as a model neurostimulant. Though photomotor responses of fathead minnows were occasionally affected by caffeine, zebrafish&#160;were markedly more sensitive for photomotor and locomotor endpoints, which responded at environmentally relevant levels. Future studies are needed to understand comparative behavioral sensitivity differences among fish with age and time of day, and to determine whether similar behavioral effects would occur in nature and be indicative of adverse outcomes at the individual or population levels of biological organization.

Here, we present a protocol to examine larval zebrafish and fathead minnow locomotor activities and photomotor responses (PMR) using an automated tracking software. When incorporated in common toxicology bioassays, analyses of these behaviors provide a diagnostic tool to examine chemical bioactivity. This protocol is described using caffeine, a model neurostimulant.

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and ...

Sampling, Sorting, and Characterizing Microplastics in Aquatic Environments with High Suspended Sediment Loads and Large Floating Debris

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only recently have microplastics in freshwater systems, such as rivers and lakes, been quantified. Microplastic sampling at the surface usually consists of deploying drift nets behind either a stationary or moving boat, which limits the sampling to environments with low levels of suspended sediments and floating or submerged debris. Previous studies that employed drift nets to collect microplastic debris typically used nets with &#8805;300 &#181;m mesh size, allowing plastic debris (particles and fibers) below this size to pass through the net and elude quantification. The protocol detailed here enables: 1) sample collection in environments with high suspended loads and floating or submerged debris and 2) the capture and quantification of microplastic particles and fibers &lt;300 &#181;m. Water samples were collected using a peristaltic pump in low-density polyethylene (PE) containers to be stored before filtering and analysis in the lab. Filtration was done with a custom-made microplastic filtration device containing detachable union joints that housed nylon mesh sieves and mixed cellulose ester membrane filters. Mesh sieves and membrane filters were examined with a stereomicroscope to quantify and separate microplastic particulates and fibers. These materials were then examined using a micro-attenuated total reflectance Fourier transform infrared spectrometer (micro ATR-FTIR) to determine microplastic polymer type. Recovery was measured by spiking samples using blue PE particulates and green nylon fibers; percent recovery was determined to be 100% for particulates and 92% for fibers. This protocol will guide similar studies on microplastics in high velocity rivers with high concentrations of sediment. With simple modifications to the peristaltic pump and filtration device, users can collect and analyze various sample volumes and particulate sizes.

Most microplastic research to date has occurred in marine systems where suspended solid levels are relatively low. Focus is now shifting to freshwater systems, which may feature high sediment loads and floating debris. This protocol addresses collecting and analyzing microplastic samples from aquatic environments that contain high suspended solid loads.

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only ...