INVERTEBRATE COMMUNITY SAMPLING PROTOCOL FOR STREAM MONITORING SITES

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Biologial Monitoring Program I. PURPOSE INVERTEBRATE COMMUNITY SAMPLING PROTOCOL FOR STREAM MONITORING SITES To desribe methods used in the olletion of stream invertebrates for the purpose of developing biologial riteria used in assessing water quality. II. REFERENCES A. Soure Douments U.S. Environmental Protetion Ageny (USEPA). 1994. Environmental Monitoring and Assessment Program - Surfae Waters and Region 3 Regional Environmental Monitoring and Assessment Program: 1994 pilot field operations and methods manual for streams. U.S. Environmental Protetion Ageny, Environmental Monitoring Systems Laboratory. Cininnati, OH. EPA/620/5-94/004. Barbour, M. T., J. Gerritsen, and J. S. White. 1996. Development of the Stream Condition Index (SCI) for Florida. Florida Department of Environmental Protetion, Tallahassee, Florida. 105 pp. B. Other Referenes U.S. Environmental Protetion Ageny (USEPA). 1996. Biologial Criteria: Tehnial Guidane for Streams and Small Rivers. Revised Edition. Offie of Water, Washington DC. EPA/822/B- 96/001. U.S. Environmental Protetion Ageny (USEPA). 1997. Revision to Rapid Bioassessment Protools for Use in Streams and Rivers (Draft). Offie of Water, Washington D.C. EPA/841/D- 97/002. III. SCOPE/LIMITATIONS This proedure applies to all site visits in whih stream invertebrates are to be olleted for the development of biologial riteria and/or the assessment of water quality. wq-bsm3-08

IV. DEFINITIONS Integrated monitoring: A stream monitoring tehnique to assess water quality using hemial, biologial and physial indiators. Environmental Monitoring and Assessment Program (EMAP): U.S. Environmental Protetion Ageny program designed to determine the status, extent, hanges, and trends in the ondition of our national eologial resoures on regional and national sales. Biologial Criteria: Narrative expressions or numerial values that desribe the referene biologial integrity of a speified habitat. Biologial riteria are the benhmarks for judging the ondition of aquati ommunities. Qualitative Multihabitat Sample (QMH): A method of sampling invertebrates whih involves sampling a variety of invertebrate habitats, inluding the following: roky substrates, inluding riffles and runs, submerged and emergent aquati vegetation, underut banks, overhanging vegetation, woody debris, and leaf paks. Intensive Watershed Monitoring: A watershed monitoring plan designed to assess the aquati health of major watersheds through intensive biologial and water hemistry sampling. This intensive approah allows assessment of watersheds for aquati life, aquati rereation, and aquati onsumption use support of the state s streams in eah of the state s 84 major watersheds on a rotating 10 year yle. V. GENERAL INFORMATION The methods desribed herein are to be applied to all wadeable and non-wadeable streams inluded in the MPCA s integrated stream ondition monitoring program. This doument is intended to be used in onert with the douments indiated in the box below if any of the desribed situations apply. For most effiient use of time and resoures, rew leaders should be able to easily ommuniate with eah other and with rews sampling for fish to ensure safe aess to sampling loations, and to avoid dupliation of effort. Data generated from samples olleted using the desribed method an be used for any of the following reasons: 1) assessment of aquati life uses as part of intensive watershed monitoring program, 2) data supplementation to aid the stressor identifiation proess, 3) development of regional biologial riteria, 4) alibration of biologial riteria.

NOTE SOP1 - Site Reonnaissane: A site reonnaissane should be done in the fall or spring prior to sampling. SOP2 - Chemial Assessment: A hemial assessment, inluding the olletion of field and laboratory hemistry samples, should be done by the fish rews. Maroinvertebrate field rews are responsible for olleting field hemistry unless a site is only being sampled for maroinvertebrates, in whih ase a laboratory sample should also be olleted. SOP3 Minnesota Stream Habitat Assessment: A habitat assessment should be onduted by the fish rews unless a site only being sampled for maroinvertebrates, in whih ase the maroinvertebrate field rew should ondut the assessment. If a habitat assessment is to be done during the same visit as an invertebrate olletion, the invertebrate olletion should be done first. VI. REQUIREMENTS A. Qualifiations of Crew Leaders A rew leader must be a professional aquati biologist with a minimum of a Bahelor of Siene degree in biology with an aquati entomology, invertebrate zoology, fisheries, or losely related speialization, or equivalent experiene in a related field. Additionally, they should have previous professional experiene working as a field biologist, inluding sampling maroinvertebrates, and onduting habitat assessments. Field rew leaders must possess exellent map reading skills, have a demonstrated profiieny in the use of a GPS (Global Positioning System), and have good interpersonal skills for dealing with landowners, and other interested stakeholders.. B. Qualifiations of field tehniians/interns A field tehniian/intern must have at least one year of ollege eduation and had oursework in environmental and/or biologial siene. C. General Qualifiations All personnel onduting this proedure must have exellent map reading skills and a demonstrated profiieny in the use of a GPS reeiver and an orienteering ompass. Beause sites may be loated miles from the nearest vehile assessable road, it is often neessary to wade through streams and/or wetlands, anoe, or hike for long distanes to reah a site. Personnel onduting this proedure must have the physial ability to aomplish this.

II. RESPONSIBILITIES A. Field Crew Leader Ensures that data generated using this proedure meet the standards and objetives of the integrated ondition monitoring program. Carries out the proedures outlined in the ation steps. B. Tehnial personnel Carries out the proedures outlined in the ation steps, inluding maintenane and stoking of equipment, date olletion and reording. VII. QUALITY ASSURANCE AND QUALITY CONTROL Compliane with this proedure will be maintained through annual internal reviews. Tehnial personnel will ondut periodi self-heks by omparing their results with other trained personnel. Calibration and maintenane of equipment will be onduted aording to the guidelines speified in the manufaturer manuals. In addition to adhering to the speifi requirements of this sampling protool and any supplementary site speifi proedures, the QA/QC requirements for this protool are as follows: A. Control of Deviations Deviations from the proedure shall be suffiiently doumented to allow repetition of the ativity as atually performed. B. QC Samples Ten perent of all sites sampled on any given year are resampled as a means of determining sampling error. C. Verifiation The field rew leader will ondut periodi reviews of field personnel to ensure that tehnial personnel are following the proedures aording to this SOP. IX. TRAINING A. All personnel, inluding experiened staff, will reeive annual instrution from a trainer designated by the program manager. B. Training ativities will inlude instrution in the field as well as a field test to ensure that personnel an implement this proedure. C. The field rew leader will provide instrution in the field to untrained personnel, suh as interns and tehniians, to ensure they an effetively exeute this proedure.

X. ACTION STEPS A. Equipment List Ensure that all of the following items are presents before implementing this proedure: Two D-frame dipnets with 500 miron mesh nets, equivalent to Wildo, turtox design Two sieve bukets with 500 miron sieves Stream Invertebrate Visit FormEletroni or hardopy of information inluded in Stream Verifiation Form, along with aerial imagery and opies of 1:24,000 USGS topographial map highlighting stream reah. Minnesota Atlas and Gazateer (Delorme) Penils Permanent/Alohol proof marker Labeling tape Internal and External invertebrate sample identifiation labels 100% reagent alohol, adequate volume to preserve four days worth of samples, a. 10-15 gallons Waterproof notebook Chest-high waders Rain-gear Jars or bottles in whih sample is to be preserved; preferably non-breakable syntheti, minimum 1 litre apaity Box or rate to store sample bottles Canoe or Kayak if needed Bakpak B. Method The multi-habitat method entails olleting a omposite sample from up to five different habitat types. The goal of this method is to get a sample representative of the invertebrate ommunity of a partiular sampling reah, it is also to ollet and proess that sample in a time and ost effetive manner. For that reason the habitats desribed below are relatively non-speifi, being hosen to represent broad ategories rather than mirohabitats. Every broad ategory inludes numerous mirohabitats, some of whih will not be sampled. It is to the disretion of the sampler whih mirohabitats are most representative of a reah. As a general rule, sample in manner that reflets the most ommon mirohabitat of any given broad habitat ategory. The habitats to be sampled inlude: Hard bottom (riffle/obble/boulder) This ategory is intended to over all hard, roky substrates, not just riffles. Runs and wadeable pools often have suitable hard substrates, and should not be exluded from sampling. The surfaes of large boulders and areas of flat, exposed bedrok are generally quite unprodutive, avoid inluding these habitats in the sampling area if possible. This is a general rule, if a partiular stream has produtive exposed bedrok, or boulder surfaes, those habitats should be onsidered sampleable. Aquati Marophytes (submerged/emergent vegetation) Any vegetation found at or below the water surfae should be onsidered in this ategory. Emergent vegetation is inluded beause all emergent plants have stems that extend below the water surfae, serving as suitable substrate for maroinvertebrates. Do not sample the emergent portion of any plant.

Underut Banks (underut banks/overhanging veg) This ategory is meant to over in-bank or near-bank habitats, shaded areas away from the main hannel that typially are buffered from high water veloities. Snags (snags/rootwads) Snags inlude any piee of large woody debris found in the stream hannel. Logs, tree trunks, entire trees, tree branhes, large piees of bark, and dense aumulations of twigs should all be onsidered snags. Rootwads are masses of roots extending from the stream bank. Leaf Paks Leaf paks are dense aumulations of leaves typially present in the early spring and late fall They are found in deposition zones, generally near stream banks, around logjams, or in urrent breaks behind large boulders. B. Method (ontinued) Sampling onsists of dividing 20 sampling efforts equally among the dominant, produtive habitats present in the reah. If 2 habitats are present, eah habitat should reeive 10 sampling efforts. If 3 habitats are present, eah habitat should reeive 7 sampling efforts. If a produtive habitat is present in a reah but not in great enough abundane to reeive an equal proportion of sampling efforts, it should be thoroughly sampled and the remaining samples should be divided among the remaining habitat types present. A sample effort is defined as taking a single dip or sweep in a ommon habitat. A sweep is taken by plaing the D-net on the substrate and disturbing the area diretly in front of the net opening equal to the net width, a. 1ft². The net should be swept several times over the same area to ensure that an adequate sample is olleted. Eah effort should over approximately.09m² of substrate. Total area sampled is a. 1.8m². One a site reah has been found or newly established, invertebrate sampling should follow. If a habitat assessment is to be onduted and/or a hemistry sample olleted it should follow invertebrate sampling. NOTE Before leaving the vehile be sure that the following equipment is brought to the site: two d-frame dipnets, two sieve bukets, site visit form, site file, GPS reeiver, amera, sample bottles, multimeter, and hemistry sample bottles if needed. 1. Assessing Stream Habitats - Before sampling an begin, the Crew Leader and field teh must determine whih habitats are present in the reah. This should be a ooperative effort. This is done by walking the length of the stream and determining whih produtive habitats dominate the stream reah. A site visit form should be filled out during this proess or immediately following sample olletion. Ideally the stream should be viewed from the top of the stream bank, but this is generally the exeption rather than the rule. For this reason, great are must be taken to walk gingerly along the stream edge, or any streamside exposed areas. If this is not possible, stay to one side of the stream so as to disturb as little substrate as possible.

B. Method (ontinued) NOTE Sine sampling should be onduted in a downstream to upstream fashion, it will save time to start the initial visual inspetion of the stream from the upstream end of the sampling reah, and walk downstream. This will allow you to start sampling at the down stream end of the reah as soon the inspetion is ompleted. It an be diffiult to estimate total stream overage of ertain habitats due to their appearane as linear or two dimensional features. Underut banks and overhanging vegetation an appear as linear features despite their depth, while snags, woody debris, vegetation mats, and emergent vegetation an appear flat despite their three dimensional nature. For these reasons best professional judgment must be used to determine what level of effort is adequate to equal one sample effort for any given substrate. Keep in mind that this method is onsidered qualitative, rulers and grids are not neessary to effetively implement this proedure. Following are some suggestions as to how approah eah habitat for the perspetive of Hard bottom: Riffles and roky runs are basially two dimensional areas, and should be thought of as suh when trying to determine how dominant the riffle habitat is in a stream. It must be kept in mind that riffles are often the most produtive and diverse habitat in the reah, relatively speaking. The field personnel must be areful to not oversample riffles. The purpose of this method is to get a representative sample. Sampling in this habitat type is relatively simple. The D-net should be plaed firmly and squarely on the substrate downstream of the area to be sampled. If the water is shallow enough, the area diretly in front of the net should be disturbed with the hands, taking are to wash large roks off diretly into the net. If the water is too deep for this, kiking the substrate in front of the net is adequate. Wath for stoneflies and mayflies trying to rawl out of the net. Vegetation: Aquati vegetation is either ompletely submerged, mostly submerged and partially floating on the waters surfae, or partially submerged and mostly extended above the waters surfae. Things like pondweed, oontail, and milfoil tend to lump and float at the waters surfae. These types of plants should be sampled with an upward sweep of the net. If the net fills with weeds, the weeds should be hand washed vigorously or jostled in the net for a few moments and then disarded. Emergent plants suh as reed anary grass and various plants in the rush family, should be sampled with horizontal and vertial sweeps of the net until it is felt that the area being swept has been adequately sampled. Plants like floating bur reed, and water elery tend to float in long strands with the urrent. They an be floating on the surfae of ompletely submerged. These plants should be sample as emergent plants with horizontal and vertial sweeps in a downstream to upstream motion. Underut banks/ Overhanging Vegetation: Underut banks and overhanging vegetation follow the line of the stream bank. Underut banks an vary in how underut they are. An additional problem is that many banks appear underut, but when investigated prove not to be. For these reasons banks should be prodded to determine how deeply they are underut. Overhanging vegetation should be treated the same way. Sampling should onsist of upward thrusts of the net, beating the underut portion of the bank or the overhanging vegetation, so as to dislodge any linging organisms.

Snags: Snags and rootwads an be large or small, long or wide, simple or twisted masses of logs or twigs that don t have any onsistent shape. Best professional judgment must be used to determine what a sampling effort is. Approximating the amount of sampleable surfae area is a sensible method with larger tree trunks or branhes. Masses of smaller branhes and twigs must be given a best guess. Given their variable nature, there is not one best method for sampling snags. Using something like a toilet brush or kithen brush works well for large piees of wood, whereas kiking and beating with the net works best for masses of smaller branhes. The person taking the sample must determine the best method for eah partiular situation. Leaf paks: Leaf paks are simple, but messy to sample. One square foot of leaf pak surfae area that has two ubi feet of leaf underneath should be sampled near the surfae. Whereas a shallow leaf pak an be sampled in it s entirety. Sweeping to the bottom of every leaf pak ould reate a disproportionately large amount of sample volume being olleted for relatively small sample area. In most situations leaf paks will not be dominate enough to be inluded in a sample. If leaf paks are sampled, it is suggested that time be spent streamside washing invertebrates off of leaves and disarding the leaves, as a leaf pak sample an easily beome overwhelmingly large. B. Method (ontinued) 2. Sampling the maroinvertebrates - After the number of produtive sampleable habitats have been determined, the sampling team should proeed in a downstream to upstream manner, sampling the various habitats present. NOTE In order to get omplete samples, the ontents of the D-net should be emptied into a sieve buket frequently. This prevents the bak flow of water resulting from a logged net. In larger streams it is onvenient for eah sampler to have a sieve buket. This allows samplers to sample independent of eah other, avoiding frequent stream rossings whih an alter the stream bed. NOTE While sampling it may beome neessary to lean the sample of muddy, fine sediment. This an be done by filling the sieve buket with lean water and allowing the resulting muky water to drain. Care must be taken not twist and turn the buket to muh, this reates a washing mahine ation whih separates insets from their deliate parts quite effetively. 3. Preserving the sample - One sampling is omplete the sample material should be preserved as quikly as possible. Transfer the sample material from the sieve buket to the sample ontainers. Sample ontainers should ontain no more than 30% of their volume as wet weight. Fill sample ontainers with 100% reagent alohol to a level that ensures a final alohol onentration of at least 70%. Be sure to thoroughly lean the buket and sampling nets of all invertebrates. The use of foreps might be neessary to dislodge some of the smaller organisms. 4. Labeling the sample Fill out internal and external sample labels for eah sample ontainer using preprinted sample labels (see attahement). Be sure to use water and alohol proof writing medium.

XI. REQUIRED RECORDS Stream Invertebrate Visit Form A. The Stream Invertebrate Visit Form should be filled out during the streamside survey, or notes should be taken on field note books and transferred to visit forms. This information will be plaed in the biologial database.

MPCA Stream Monitoring Program STREAM INVERTEBRATE VISIT FORM Stream Name: Date: Field Number: County: Crew: Water Chemistry Tape Down:. (1/100ths ft) Loation: Time: (24 hr) : Air Temp: ( o C) Water Temp: ( o C) Condutivity: (umhos@25 o C) DO: (mg/l) DO % Saturation: ph: Sehi -Tube: (m) Water Level: Normal Below (m) Above (m) Color (pu) ***If Flagging is not found or if establishing a new site, fill out GPS info*** Coordinates LATITUDE LONGITUTDE Time: Field GPS:.. Name: Notes: Flow Habitat Substrate Stream Classifiation Information Flow over riffle(s) High / Med / Low / NA Exavated, trapezoidal hannel % Flow at reah onstrition High / Med / Low / NA Shallow exavation, hannelized wetland % Flow over run High / Med / Low / NA Natural hannel % General flow pattern High / Med / Low / NA Emergent, aquati vegetation in hannel Ext / Mod / Sparse / NA Intermittent setions Yes / No Emergent, aquati vegetation along bank Ext / Mod / Sparse / NA Riffle (with flow) present in reah Floating or submerged aquati vegetation Ext / Mod / Sparse / NA Riffle (with flow) present outside of reah Loosely attahed filamentous algae Ext / Mod / Sparse / NA (riffles do not inlude riprap assoiated with bridges or bank stabilization) Firmly attahed algae or submerged veg Ext / Mod / Sparse / NA Dominant invertebrate habitat (irle two) Riffle Roky Run-Pool Aquati Marophyte Bank-Overhanging Veg Wood Leaf Dominant Run Substrate bedrok / boulder / obble / gravel / sand / silt Dominant Pool Substrate bedrok / boulder / obble / gravel / sand / silt Dominant Substrate reeiving flow bedrok / boulder / obble / gravel / sand / silt Dominant Substrate in reah bedrok / boulder / obble / gravel / sand / silt Channel Vegetation Stream displays a typial riffle-run pool morphology adequate flow to maintain riffle organisms inadequate flow to maintain riffle organisms Stream has adquate flow to maintain riffle organism, but does not have suitable oarse substrate to support these assemblages (riffles, rok substrate in runs or pools) Stream has adquate flow to maintain riffle dwelling organism, woody debris has replaed roks as primary oarse substrate Stream is low gradient, stream bed is predominately fine substrate, inadequate flow to maintain riffle organisms Invertebrate Sample Information Qualitative Multi-Habitat Sample (QMH) Divide 20 samples equally among habitat types present in the reah. If three habitat types are present take 7 samples in eah of the three dominant habitats (for a total of 21). If a habitat is present, but not in abundane to sample in equal proportion to other habitats, sample as muh as possible and divide the remaining samples between the dominant habitat types. a Habitat # rok riffle/run Flow adequate to arry insets into net rok substrate Artifiial flow needed to arry inset i Additional Biologial Information Presene of freshwater sponge ----------- yes / no Presene of exoti speies ---------------- yes / no Name of exoti(s) if present: (vouher a speimen if not present in sample) Presene of mussels ----------------------yes / no Desription of mussel density and/or mussel bed loation: aquati marophyte Notes underut bank overhanging veg snag woody debris root wad leaf pak Number of multihabitat ontainers: Pitures #: DD DU MD MU UD UU

Stream Sample External Label MPCA Bioassassment Invertebrate Sample Sample Preservative - 100% reagent alohol / 10% formalin Sample Type: QMH / RTH Sample Composition: Riffle / Bank / Wood / Veg Date / /20 (mm/dd/yyyy) Station Name Station ID Site Visit 1 / 2 Sample Jar of Colletors Stream Sample Internal Label Invertebrate Sample sample type Site Name: Field Number Date: / / Bottle No. of Colleted by: