Multi-habitat (MHAB) Macroinvertebrate Collection...

Multi-habitat (MHAB) Macroinvertebrate Collection Procedure

S-001-OWQ-W-BS-10-T-R0 Technical Standard Operating Procedure

Office: Office of Water Quality Branch: Watershed Planning and Assessment Branch

Section: Biological Studies Section

Revised: N/A Revision Cycle: Every 2 years Effective date: October 30, 2010

Scope of operations

The purpose of this technical standard operating procedure (TSOP) is to ensure that the sampling of benthic macroinvertebrate communities using the Multi-Habitat (MHAB) macroinvertebrate sampling method is completed in a consistent manner.

Scope of applicability

This TSOP applies to Indiana Department of Environmental Management (IDEM) Office of Water Quality (OWQ) Biological Studies Section (BSS) staff who collect macroinvertebrate community samples from rivers and streams as part of the Indiana Department of Environmental Managements (IDEM’s) ambient water quality monitoring program. Authorizing Signatures

Multi-habitat (MHAB) Macroinvertebrate Collection Procedure S-001-OWQ-W-BS-10-T-R0

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Table of Contents 1.0 Overview “Work-flow” Flowchart ...................................................................................................... 3 2.0 Definitions ........................................................................................................................................ 3 3.0 Roles ................................................................................................................................................ 4

3.1. Responsibilities: ........................................................................................................................... 4 3.2. Training and Experience Requirements: ..................................................................................... 5

4.0 Description of Equipment, Forms, and/or Software to be Used....................................................... 6 5.0 Procedure ......................................................................................................................................... 8

5.1. Procedural Flowchart ................................................................................................................... 8 5.2. Procedure .................................................................................................................................... 9 5.3. Related Technical Issues ........................................................................................................... 13

5.3.1. Health and Safety Warnings ...................................................................................................... 13 5.3.2. Cautions ..................................................................................................................................... 14 5.3.3. Interferences .............................................................................................................................. 14 5.3.4. Calibration .................................................................................................................................. 14 5.3.5. Troubleshooting ......................................................................................................................... 14

6.0 Records Management .................................................................................................................... 14 7.0 Quality Assurance / Quality Control ............................................................................................... 15 8.0 References ..................................................................................................................................... 15 9.0 History of Revisions ....................................................................................................................... 16 10.0 Appendices .................................................................................................................................... 16


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1.0 Overview “work-flow” flowchart

Project Generation

Initial Site Recon

Health and Safety

Training

Travel to Site

Complete Stream Sampling Datasheet with explanation as to why site was not sampled

In-situ Water Chemistry

Biological Sampling of Macroinvertebrate

Community Assemblage

QHEIReturn to Laboratory

Site Sampled?

Time for another

sample?

No

Yes

No

Yes

Take photos of site

2.0 Definitions 2.1. “AA/AB Number” – Number assigned to each individual watershed sampling event conducted

by Indiana Department of Environmental Management (IDEM) Field Crews. This number is used to identify the sampling event in the Assessment Information Management System database (AIMS data base).

2.2. "Agency staff” – Any employee or representative of the Indiana Department of Environmental Management including regular employees, temporary employees, contractors and interns.

2.3. "Ambient” – Surrounding environmental conditions.

2.4. "Assessment Information Management System database (AIMS database)" – IDEM database containing information related to water chemistry, aquatic habitat, macroinvertebrate and fish communities, fish tissue analyses, sediments and E. coli bacteria data collected by Agency staff from watershed sampling events.

2.5. "Biological Studies Section (BSS)" - A section within the Watershed Planning and Assessment Branch of the Office of Water Quality

2.6. "Deionized water" – Water that has had its mineral ions removed.

2.7. “Crew Leader” – The Agency staff person who leads a Field Crew when conducting field sampling activities.

2.8. “Field Crew” – The team of Agency staff who conduct field sampling activities. Field Crews must contain at least one full-time Agency staff member from the Biological Studies Section in the Crew Leader position and one or more fulltime IDEM employees, Governor’s Summer Interns, or uncompensated interns.

2.9. “Glide” – An area common to most modified stream channels that do not have distinguishable pool, run, and riffle habitats; the current and flow is similar to that of a canal; the water surface gradient is nearly zero (Ohio EPA 2006).

2.10. “Governor’s Summer Intern” – An intern selected under the Governor’s Public Service Internship program; a program created to introduce college students to the operations and officials of state government. Governor’s Summer Interns are compensated, intermittent employees usually working full-time hours from May to September.

2.11. "Hydrolab readings" – The measurements of water quality of both ambient and effluent waters taken from a Hydrolab.


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2.12. "Indiana Department of Environmental Management (IDEM)” – An agency of Indiana State Government whose mission is to implement federal and state regulations to protect human health and the environment while allowing the environmentally sound operations of industrial, agricultural, commercial and government activities vital to a prosperous economy.

2.13. “Laboratory Custodian” – The Agency staff person responsible for checking in and initial curation of algae, fish and macroinvertebrate samples brought into the laboratory from field sampling.

2.14. “Macroinvertebrate Sample Number” – An identification number used to identify a macroinvertebrate sample. The macroinvertebrate sample number is constructed by combining the last two digits of the year, the two digit approximation of the month, the two digit approximation of the day, the number of the field notebook used by the crew leader to record the sampling event, and a two digit number indicating the order in which the current sample was collected on that particular day. For example, the fourth sample collected by the field crew using field notebook #3 on July 15, 2010 would read: MHAB 100715304 (“10” =2010, “07” = July, “15” = 15th day of the month, “3” = field notebook #3, “04” = fourth sampled collected that day). The macroinvertebrate sample number does not take precedence over the “AA/AB Number”.

2.15. “Office of Water Quality (OWQ)” - The Office of Water Quality within the Indiana Department of Environmental Management.

2.16. "Ohio Environmental Protection Agency (Ohio EPA)” – An agency of the Ohio State Government whose mission is to protect the environment and public health by ensuring compliance with environmental laws and demonstrating leadership in environmental stewardship.

2.17. "Periphyton sampling" – The sampling of algae attached to submerged substrates in aquatic environments.

2.18. “Pool” – An area of the stream with slow current velocity and a depth greater than riffle and run areas; the stream bed is often concave and stream width frequently is the greatest; the water surface slope is nearly zero (Ohio EPA 2006).

2.19. “Riffle” – The shallow parts of the stream where water flows swiftly over completely or partially submerged pebble to boulder sized rocks to produce surface agitation.

2.20. “Run” – Areas of the stream that have a rapid non-turbulent flow; runs are deeper than riffles with a faster current velocity than pools and are generally located downstream from riffles where the stream narrows; the stream bed is often flat beneath a run and the water surface is not visibly broken (Ohio EPA 2006).

2.21. “Uncompensated (unpaid) intern” – Interns who work voluntarily and do not receive monetary compensation for their work at IDEM, but often receive college credit hours from the accredited university in which they are enrolled.

3.0 Roles 3.1. Responsibilities:

The “Crew Leader” is the IDEM Biological Studies Section staff person who leads a team of IDEM employees when conducting macroinvertebrate field sampling activities. The crew leader is responsible for collecting the MHAB macroinvertebrate sample, filling out field paperwork and verifying that tasks performed by the remaining field crew members are completed correctly. The “Field Crew” consists of the members of the team of IDEM Biological Studies Section staff persons who conduct field sampling activities. The field crew (minus the crew leader) is responsible for conducting field water chemistry data collection, habitat measurements, and any other field assessment activities as designated by the crew leader. A field crew for a macroinvertebrate sampling event usually consists of two staff persons: a crew leader who collects the macroinvertebrate sample and another crew member who is responsible for


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obtaining Hydrolab readings, densiometer measurements and stream width measurements. However, a staff person who normally functions as a crew leader and collects macroinvertebrate samples may perform this function while also working as a crew member on a fish or periphyton sampling field crew. The “Laboratory Custodian” is the IDEM Biological Studies Section staff person responsible for receiving and initial curation of algae, fish and macroinvertebrate samples collected during field sampling activities. The laboratory custodian checks that the information written on sample labels, in the field notebook and in the Biological Sample Chain of Custody form (Appendix 10.1) is correct.

3.2. Training and Experience requirements: The crew leader of the macroinvertebrate sampling team will usually be the person who conducts the MHAB macroinvertebrate sampling procedure. This person will typically have had the most experience conducting macroinvertebrate sampling, particularly with the MHAB sampling method. The crew leader may choose to conduct on-site training with other field crew members to help them reach an acceptable level of proficiency with the MHAB sampling method (with the ultimate goal of being able to collect a MHAB sample that is comparable to a MHAB sample collected by the crew leader). Presently though, only MHAB samples collected by the crew leader will be used for water quality assessment purposes. As the remainder of the field crew becomes more proficient with the MHAB method, they may be expected to collect MHAB samples for assessment purposes. It is recommended that field staff conducting MHAB macroinvertebrate sampling should have previous experience with the identification of aquatic macroinvertebrates. This experience may be gained through college courses that feature the study of aquatic macroinvertebrates as part of the curriculum (e.g., Entomology, Invertebrate Zoology or Limnology), attendance at workshops focusing on aquatic macroinvertebrate identification, pre-IDEM job related experience (conducting aquatic macroinvertebrate sampling and/or laboratory processing with a consulting firm or a water quality agency), on-site training by experienced IDEM staff or by time spent processing aquatic macroinvertebrate samples in an IDEM laboratory. It is important that field staff conducting MHAB macroinvertebrate sampling be able to recognize by visual observation all orders and most families of aquatic macroinvertebrates that are likely to be encountered in order to collect a sample that is representative of the macroinvertebrate community composition at that location.

All field staff are required to have an IDEM-approved 24 hour initial health and safety training course and thereafter, an IDEM-approved 8 hour refresher course annually to maintain field

status (IDEM 2007a). Until initial health and safety training is received by new employees or temporary staff, they must be accompanied in the field by a staff member who is approved for field status and has taken the initial 24 hour health and safety training.

Biological Studies Section field personnel will comply with the Biological Studies Section (BSS) Standard Operating Procedures Manual: Section II. Hazard Communications Manual (IDEM 1992) which includes four, yellow, 3-ring binders consisting of 1) BSS Safety Manual 2) BSS Hazard Communication and Standard Operating Procedures 3) Occupational Safety and Health Administration Handbooks 4) Material Safety Data Sheets.

Sampling on surface waters requires safety consciousness of staff members. If an injury or illness arises in the field, staff will follow the IDEM Policies and Procedures entitled "Injury and Illness Standard Operating Procedures" (IDEM 2007b). Operating in and around waterbodies carries inherent risks of drowning; thus, personnel involved in sample collection will wear appropriate clothing and personal protective equipment (IDEM 2008) when operating boats or sampling in deep water or swift currents. According to the memorandum “Use of Personal Flotation Devices (PFDs) by Branch Personnel” dated February 29, 2000, staff must wear USCG approved Type I, II, or III PFDs whenever

the planned work requires them to enter the water and the maximum water depth at any place at the work site is over their knee (note that this depth depends on the


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employee but it will usually be between 12 and 20 inches or 300-500 mm) or the employee is in a watercraft of any kind that is being launched, is in the water, or

is being retrieved from the water or the employee must work from structures that do not possess guard rails and are over

or alongside water where the water depth is or could reasonably be expected to be 3 feet deep.

In addition, when work is being done in boats on co-jurisdictional waters (as defined by Indiana Code § 14-8-2-315) or during hours of darkness on any waters of the state, all personnel in the watercraft must wear a high intensity whistle and SOLAS certified strobe light. All questions or concerns of safety issues arising in the field should be brought to the attention of the field crew leader.

4.0 Description of equipment, forms, and/or software to be used 4.1. “Denatured Ethanol (70% ethanol)” – An aqueous mixture composed of 70% 200 Proof

Reagent Alcohol (89.5-91.5% ethanol (Cas No. 64-17-5), 4.0-5.0% methanol (Cas No. 67-56-1) and 4.5-5.0% Isopropyl alcohol (Cas No. 67-63-0) (Ultra Pure 2009)) and 30% deionized water, prepared in the lab prior to field sampling. Found in the flammable liquids cabinet in Room 124 at IDEM Shadeland office (Fish processing lab). The 70% ethanol is used by the crew leader as the primary preservative for the macroinvertebrate sample.

4.2. “Alcohol Safety Can” – Metal container with spring closing lid, spout cover and flash

arresting screen used to store and transport 70% ethanol. Found in Room 125 at IDEM Shadeland office (Macroinvertebrate processing lab). Used by the field crew to safely transport 70% ethanol during field activities.

4.3. “Backpack” – External frame backpack. Found in Room 125 at IDEM Shadeland Office.

Used by the field crew to transport macroinvertebrate sampling equipment to sampling site.

4.4. “Biological Studies Field Chain of Custody form (chain of custody form)” – Form used to

record samples collected during a biological stream sampling event (see Appendix 10.1). Used by the crew leader to record the macroinvertebrate sample number, AA/AB number, type of sample container, and date and time sample was collected.

4.5. “D-frame Net” – Macroinvertebrate sampling net consisting of a wooden 52 inch handle

attached to a 1 foot x 1 foot D-shaped frame fitted with a net with a mesh size of 500 micrometers. Found in Room 125 at IDEM Shadeland office. Used by the crew leader to conduct MHAB macroinvertebrate sampling.

4.6. “Digital Camera” – Kodak EasyShare four megapixel digital camera. Found in the

electronics storage cabinet in Room 125 at IDEM Shadeland office. Used by field crew to take upstream and downstream pictures of sampling site.

4.7. “Face Shield” – Clear plastic face shield. Found in Room 125 at IDEM Shadeland office.

Used by the crew leader when handling formalin to prevent possible injury.

4.8. “Field Notebook” – Nalgene field notebook, 9.25” by 11.25”, 48 pages, with blue polyethylene cover and coated Polypaper pages. Found in Room 125 at IDEM Shadeland office. Used by crew leader to record site information for each sample collected (see Appendix 10.2)

4.9. “Forceps” – 4A or “fine-tip” forceps. Found in Room 125 at IDEM Shadeland office. Used

by the crew leader to pick macroinvertebrate specimens from the sample tray and deposit them into the sample jar.

4.10. “Formalin” – An aqueous mixture (Cas No. 82115-62-6) composed of 37% formaldehyde,

10-15% methanol and 48-53% de-ionized water, diluted in the lab to a 10% solution with


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de-ionized water and stored in a 1-liter squeeze and pour dispensing bottle. Found in the flammable liquids cabinet in Room 124 at IDEM Shadeland office. The crew leader adds a small amount of formalin to the 70% ethanol in the sample container as an additional preservative for the macroinvertebrate sample. See Newhouse 1998 for instructions on the safe handling and usage of formalin.

4.11. “Global Positioning Satellite (GPS) Unit” – Garmin eTrek Legend GPS units used for

navigation. Found in the electronics storage cabinet in Room 125 at IDEM Shadeland office. Used by field crew to navigate to the sampling site.

4.12. “Hydrolab” – Multiprobe Hydrolab Quanta unit used to measure basic water chemistry

parameters. Found in Room 125 at IDEM Shadeland office. Used by the field crew to measure field chemistry parameters at a sampling site.

4.13. “Measuring Tape” – Keson Open Reel Fiberglass Measuring Tape, 50 meters (165 feet)

in length. Found in Room 125 at IDEM Shadeland office. Used by field crew to measure width of the stream and the 50 meter zone used for the sweep portion of the MHAB sample.

4.14. “No. 35 Sieve” – No. 35 U.S. Standard Testing Sieve, mesh size of 500 micrometers

(0.0197 inches). Found in Room 125 at IDEM Shadeland office. The crew leader pours the contents of the sample bucket through the sieve to separate the macroinvertebrate specimens from the water in the sample.

4.15. “Office of Water Quality Macroinvertebrate Header form (macroinvertebrate header form)”

– Form used to record general habitat information during a biological stream sampling event (see Appendix 10.3). Forms are individually generated for sites by AIMS. Used by the crew leader to record basic observations on stream morphology, substrate composition and riparian zone features conditions.

4.16. “Office of Water Quality Biological Studies QHEI (Qualitative Habitat Evaluation Index)

form (QHEI form)” – Form used to record specific habitat information during a biological stream sampling event (see Appendix 10.4). Forms are individually generated for sites by AIMS. Used by the crew leader to record qualitative observations on stream substrate composition, instream cover, channel morphology, bank erosion, riparian zone quality, and pool/glide and riffle/run quality.

4.17. “Office of Water Quality Original Photographic Chain of Custody form (photo chain of

custody form)” – Form used to record photographs taken during a biological stream sampling event (see Appendix 10.5). Used by the crew leader to record the site name (including stream name and Corvalis and AIMS L-Site numbers), number of photos taken at a site and photographer name.

4.18. “Rangefinder” – Bushnell Laser Rangefinder Yardage Pro Scout. Found in electronics

storage cabinet in Room 125 at IDEM Shadeland office. Used by field crew to measure width of the stream and the 50 meter zone used for the sweep portion of the MHAB sample.

4.19. “Sample Bucket” – Rubbermaid 2963 Round Bucket, 10 quart capacity, with handle and

pouring spout. Found in Room 125 at IDEM Shadeland office. Used by the crew leader to store riffle kick and 50 meter sweep components of the MHAB sample until elutriation.

4.20. “Sample Container” – Nalgene 250 ml clear plastic jar with white lid. Found in Room 125

at IDEM Shadeland office. Filled with 70% ethanol and used by the crew leader to store the collected macroinvertebrate sample until the sample can be processed.

4.21. “Sample Label” – Label printed on waterproof paper that is inserted into a sample

container at the time of collection of the sample. Label is printed prior to field sampling


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and contains pertinent information such as the stream name, county name, nearest road, and AA/AB Number. At the time a sample is collected, the date, initials of the collector, and MHAB Number are written on the label before it is inserted into the sample container.

4.22. “Sample Tray” – White plastic sample tray, 18 inches x 14 inches x 3.5 inches. Found in

Room 125 at IDEM Shadeland office. Used by the crew leader to hold contents of sieve after elutriation and retain macroinvertebrate specimens during the pick process.

4.23. “Stream Sampling Field Data Sheet (stream data sheet)” – Form used to record general

water chemistry information during a biological stream sampling event (see Appendix 10.6). Forms are individually generated for sites by AIMS. Used by the crew leader to record basic observations on stream appearance, water chemistry parameters and meteorological conditions.

5.0 Procedure 5.1. Procedural Flowchart

Is there time to sample

another site?

Locate riffle/run to be

sampled

Determine 50m of shore

to be sampled

Conduct riffle kick

Conduct shoreline

sweep

Remove large debris from sample

Elutriation of sample

15 minute sample pick

Complete sample label

Macroinvertebrate Header Form

QHEI

Chain of custody form

Field notebook

entry

Add formalin to sample

Return to office

Check in macroinverterbrate

samples

Begin:Drive to

sampling site

Put sampling items in

backpack

Navigate to x-point

Can site be sampled?

Complete appropriate

forms and take site photo

Is there time to sample

another site?

No

Yes

No

NoYes

Yes

In-situ water chemistry

Site photo


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5.2. Procedure

5.2.1. Using hand held GPS units, Indiana gazetteers and other site-specific maps (Appendix 10.7, 10.8) the field crew will drive to a sites designated parking location as described in the site reconnaissance form (Appendix 10.9).

5.2.2. The crew leader will fill three sample containers two-thirds full with 70% ethanol from

the alcohol safety can. The crew leader, using the checklist provided in Appendix 10.10, will check to make sure that all of the equipment necessary to conduct macroinvertebrate sampling is present and stored in the backpack for transportation to the probabilistic sampling site (the “x-point”).

5.2.3. Using hand held GPS units and the site reconnaissance form the field crew will walk to

the x-point. Upon arrival at the x-point the crew leader will determine if the site can be sampled. In order to be sampled, water must be present in 50% of the stream reach, defined as 15 times the wetted width of the stream (IDEM 2005). If the site can be sampled the crew leader will select an area of the stream shore to serve as a sample processing location and a place to leave equipment that is not currently in use. If the site cannot be sampled the crew leader will fill out the following portions of the stream data sheet,

Sample Collectors: Three letter initials of the crew leader and all members of the field crew. Date: Date site was visited, entered as MM-DD-YYYY. Time: Time sample was collected, entered in a 12-hour time format. Sample Taken?: Select the appropriate reason the sample was not collected (“Stream Dry”, “Owner Refused Access”, “Frozen”, “Other”). If “Other”, describe the reason the sample could not be collected in the “Special Notes” area of the stream data sheet. The crew leader will also fill out an entry for the site in the field notebook with the reason the

site was not sampled, take upstream and downstream pictures of the site, and return to the vehicle and travel to the next site, if time allows.

5.2.4. The crew leader will visually survey the in-stream and surrounding riparian habitat of

both shores of a stream reach 50 meters upstream and 50 meters downstream of the x-point. These 50 meter areas of the stream should be accurately measured using a 50 meter measuring tape or a rangefinder. Ideally, a good quality, wadeable riffle would be located within 50 meters of the x-point; if this is not the case, then a riffle located no more than 15 times the streams wetted width (to a maximum of 500 meters) from the x-point may be sampled. If a riffle is not available, the crew leader should locate a good quality wadeable run or glide habitat to be sampled during the kick portion of the MHAB sampling procedure. A 1-minute kick sample is not collected in deep U-shaped (or bowl shaped) streams and in streams where low-flow has reduced the stream to interstitial flow between a series of pools; in these instances the 50m MHAB sweep by itself is sufficient.

Using best professional judgment, the crew leader will then determine the 50 meter section of stream shoreline habitat that will potentially yield the most diverse macroinvertebrate sample (the quality of the sample can be improved by selecting the section of stream featuring the largest number of productive in-stream macroinvertebrate habitat types, avoiding collecting samples downstream of bridges and tributaries flowing into the stream and attempting to collect the sample from an area that contains a riffle-run-pool complex). The x-point (which is fixed and must, under normal circumstances, be included in the macroinvertebrate sample) may be located at the “top” of the sampling reach (by sampling a 50 meter stretch of one shore downstream of the x-point), at the “bottom” of the sampling reach (by sampling a 50 meter stretch of one shore upstream of the x-point), or may be in the middle of the sampling reach (for instance, sampling 20 meters upstream and 30 meters downstream of the x-point on one shore). If necessary, the sampling reach may also be defined as two alternating or parallel sections of shore on opposite banks that, combined, total 50 meters and includes the x-point.

If the actual x-point is located on a section of the stream that is currently dry or inaccessible, then the sample may be collected in a section of the stream containing water that is within 15 times the wetted width of the stream from the x-point. This adjustment to the sampling method


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should be used with reservation and only if the resulting sample location will provide similar habitat to what would have been found at the actual x-point. The reason for such a modification and the area of the stream sampled should be described in the field notebook and on the drawing of the site made in the QHEI form.

5.2.5. In the riffle kick portion of the MHAB sample the crew leader will locate an area in the

lower (downstream) area of the riffle or run habitat that is to be sampled. While holding the d-frame net handle, the flat end of the d-frame net is placed securely onto the substrate so that there are no gaps under the frame. The crew leader will disturb (by kicking) the riffle/run substrate one foot directly in front of the d-frame net so that any dislodged macroinvertebrates are carried by the current into the net. The crew leader moves in an upstream or diagonal upstream direction across the riffle/run, continuously disturbing the substrate in front of the d-frame net, for a distance of about 10 feet (this is roughly equivalent to sampling one square meter of riffle/run habitat). The riffle kick portion of the MHAB sample is timed and should be completed in 60 seconds (time may be kept by the crew leader or, preferably, by another crew member using a stopwatch). After completion of the kick, the crew leader should inspect the contents of the net. Any stones (gravel sized or smaller) or debris (leaves, small sticks) that are kicked into the net are retained. Cobble and large sticks kicked into the net should be visually inspected and any attached macroinvertebrates removed and placed in the net; the cobble or large sticks are then returned to the stream. Streams with little or no flow may require the crew leader to disturb the substrate and sweep the d-frame net through the disturbed water column above the substrate to collect this portion of the sample. After completion of the kick the contents of the d-frame net are emptied into a plastic bucket filled half full with water. This bucket should be left at the sample processing location to reduce the risk of losing the macroinvertebrate sample and may be covered to prevent the escape of any flying species (mostly adult Coleoptera and Hemiptera species).

5.2.6. The sweep portion of the sample begins with the crew leader at the lower

(downstream) portion of the 50 meters of shoreline habitat that is to be sampled. The d-frame net is swept over the shoreline habitat so that the net dislodges any macroinvertebrates on the substrate surface and pushes them into the water column. The d-frame net is then swept through this disturbed area above the substrate two or three times to collect any dislodged macroinvertebrates. Sweeps are conducted at roughly 1.5-2 meter intervals for the entire length of the 50 meters of shoreline habitat for a total of about 25-30 sweeps. Using best professional judgment and experience, the crew chief should sample as many microhabitats (rootwads, emergent vegetation, woody debris, leaf packs, sandy and silty substrates, and cobble and gravel substrates) as possible, sampling these microhabitats in relative proportion to their occurrence in the 50 meters of shoreline habitat. Care should be taken to avoid collecting large amounts of debris in the net as this will slow down the elutriation and picking processes. After completion of the 50 meter habitat sweep, the crew chief empties the contents of the net into the same bucket with the contents of the kick component of the sample.

5.2.7. The crew leader removes and visually inspects any cobble, large gravel, woody debris and large leaves that were emptied into the bucket (during this and the following portion of the sample, the crew leader should wear elbow length rubber gloves to minimize the risk of injury, see section 5.3.1). Any macroinvertebrates found on these items are removed with forceps and placed into the white plastic tray. After inspection, these debris items are discarded. Due to the conservation status of many species, any freshwater mussels of the family Unionidae collected in the sample should be returned to a suitable area of the stream as soon as possible (their presence and species, if known, should be recorded on the stream data sheet and in the field notebook).

5.2.8. After removal of large items, the crew leader will use their arm to swirl the contents of

the bucket and then pour the water through a No. 30 sieve. The water flow from the bucket should be continuous but stopped before any sand is poured into the sieve. After each pour, the contents of the sieve are emptied into the white plastic tray. The bucket containing the sample is then refilled with stream water using a separate bucket (taking care to not add any additional macroinvertebrates via the refill bucket). This process is repeated until the water poured through


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the sieve is clear and free of silt, a minimum of five times. After the elutriations, the crew leader will inspect the contents of the bucket and remove any heavier organisms that were not emptied into the sieve (e.g. crayfish, snails, and Asian or fingernail clams). Handfuls of any remaining sand, leaves or fine gravel are then removed, visually inspected for any remaining organisms, and discarded.

. 5.2.9. The objectives of the pick are to collect a sample that contains the greatest diversity of

organisms and also reflects the relative abundance of organisms in the entire sample. The crew leader uses a pair of forceps to individually pick macroinvertebrates from the contents of the tray for 15 minutes. Picked organisms are deposited into a sample container two-thirds filled with 70% ethanol. A second container may be used for crayfish or dobsonflies if they are numerous in the sample (these organisms are usually large and take longer to expire; as a result they can damage or destroy smaller, fragile specimens in the sample container). Another member of the field crew may use a stopwatch to time the pick for the crew leader and announce when the pick time remaining reaches 10, 5 or 1 minute(s).

5.2.10. After completion of the 15 minute pick, the crew leader fills out a sample label and

inserts it into the macroinvertebrate sample jar(s). These labels should be pre-printed with the AA/AB Number, stream name, county, distance from nearest road or town, and latitude and longitude. Using a lead pencil, the sample collector’s initials, date of collection, and the Macroinvertebrate sample number are added to the sample label. If a duplicate sample is collected or if multiple sample containers are used, additional sample labels can be made in the field using a small piece of Rite-in-the-Rain paper, and should include all of the previously listed types of information. If multiple sample containers are used for a single sample, the labels should also include “1 of 2” and “2 of 2“.

5.2.11. While the crew leader is completing the MHAB macroinvertebrate sample, other

members of the field crew should complete the Hydrolab sample of field water chemistry and enter this information into the stream data sheet. They should also take measurements of the stream width (recorded in the macroinvertebrate header form), conduct densitometer measurements of the canopy cover (entered onto the back of the QHEI form (Appendix 10.4)), and take upstream and downstream pictures of the sampling site (recorded in the photo chain of custody form). These forms should be checked by the crew leader before leaving the site.

5.2.12. Upon completion of the MHAB macroinvertebrate sample the crew leader will

complete the following portions of the macroinvertebrate header form: General Site Information Sample #: The AA/AB sample number. Should be automatically filled out by AIMS prior to sampling. Macro #: The nine-digit Macro Sample Number (see Section 2.13 for instructions on constructing this number). Macro Sample Type: Currently, the MHAB sampling method is not an option on this form. “MHAB” should be written in this area underneath the entry for “CPOM”. # Containers: The number of containers used at the site to contain the sample. Notes: Any notes pertinent to the sample or site conditions when sample was collected. Stream Name/Location: The name of the stream and the road nearest to the stream. Should be automatically filled out by AIMS prior to sampling. County: County where sample is collected. Should be automatically filled out by AIMS prior to sampling. Sample Date: Date when sample was collected. Riparian Zone/Instream Features Erosion: Check one to provide an estimation of the severity of bank erosion in the sampling reach. NPS Pollution: Check one to provide an estimation of any potential non-point sources of pollution in the sampling reach. Stream Depth: Enter an estimation (in meters) of the average riffle, run, and pool depths, if


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they were present at the site. If these stream features are not present, leave blank. Distances: Enter an estimation (in meters) of the distance between riffles or bends if more than one riffle or bend are present within the stream reach visible from the x-point. If one or no riffles or bends are present, leave blank. Predominant Surrounding Land Use: Check one to provide an estimation of the predominant land use in the area surrounding the sampling reach. If the stream has been channelized or if a dam is present, check the appropriate box. Stream Width: Enter (in meters) the width of the stream at the x-point. High Water Mark: If there is evidence of recent high flow (debris collected in tree limbs, traces of mud left on riparian vegetation) enter (in meters) an estimation of the maximum height of flow, above the current stream level. Canopy Cover (% Open): Enter an estimation of the percent of stream area in the sampling reach not shaded by riparian vegetation. Sediment Sediment Odors: Check any odors that were detected during the sampling event. Sediment Deposits: Check any sediment deposits that were encountered in the sampling reach. Sediment Oils: Check one to provide an estimation of sediment oils (if any) that were encountered during the sampling event. If the undersides of un-embedded stones are black, check the appropriate box. Substrate Components Inorganic Substrate Components: Enter in percentages of multiples of 5 (0%, 5%, 10%, etc.) the amount of substrate composed by Bedrock, Boulders, Cobble, Gravel, Sand, Silt or Clay. Organic Substrate Components: Enter in percentages of multiples of 5 (0%, 5%, 10%, etc.) the amount of substrate composed by Detritus (as sticks or wood), Detritus (as CPOM), Muck/Mud or Marl. Water Quality Water Odors: Check any water odors that were detected during the sampling event. Water Surface Oils: Check any water surface oils that were detected in the sampling reach. Stream Type: Check whether the stream was warm or cold water. Criteria for this designation can be found in Aquatic Research Center of the Indiana Biological Survey 2007. Turbidity: Provide an estimation of the streams turbidity in the sampling reach.

5.2.13. The crew leader will complete the QHEI for the site following instructions described by Sobat 2006 and Ohio EPA 2006. Macroinvertebrate sample number, AA/AB Number, Number of Sample Containers, and Date and Time Collected, will then be filled out on the biological samples chain of custody form. The crew leader will also fill out an entry in the field notebook listing the same information found on the sample labels, whether the riffle portion of the sample was collected, the side of the stream bank where the 50 meter sweep sample was collected and its position relative to the x-point, and any other pertinent information. If any modifications were made to the sample method, they should be described here. 5.2.14. After checking to ensure that all samples have been properly labeled and all site forms have been completed, the field crew will store all of the equipment used to complete the sample in the backpacks, leave the site and navigate to the field vehicle. Upon arrival at the field vehicle the crew leader will add 2-4 ml of formalin to the sample containers, depending on the amount of organic material in the sample container (the crew leader should wear appropriate personal protection and exercise extreme caution when handling formalin, see section 5.3.1). The crew leader will then affix a piece of white tape to the lid of the sample container listing the stream name, AA/AB Number, and Macroinvertebrate sample number. 5.2.15. The crew leader will determine if there is sufficient time to complete another sample and, if there is, the crew will continue to the next site. If the crew leader determines that there is


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not enough time to complete another sample, the field crew will return to their hotel or to the IDEM Shadeland branch office. 5.2.16. Upon arrival at the IDEM Shadeland branch office, the macroinvertebrate samples collected by the field crew will be brought into Room 121 for temporary storage. The crew leader will include their signature at the top of the biological samples chain of custody form to certify that the samples were collected by them or in their presence, enter the date the samples were brought into the IDEM Shadeland branch office, and circle the type of samples that were collected (“macro”). The lab custodian will check to make sure that the information on the sample labels inside the sample containers matches the information entered onto the biological samples chain of custody form. The lab custodian will then enter the date and time the samples were brought into the IDEM Shadeland branch office, the storage room number, and a check to certify that the sample was present and accounted for. The crew leader and lab custodian will complete the form by signing their names in the appropriate “Relinquished by:” and “Received by:” areas and entering the time and date that the samples were signed over. The samples are then placed in Room 125 until they are ready to be processed.

5.3. Related Technical Issues

5.3.1. Health and Safety Warnings All field crew members must wear USCG approved Type I, II, or III PFDs whenever

there is the possibility that water over the knee in depth may be encountered. Failure to do so may result in injury or drowning.

All field crew members should wear neoprene chest waders and felt-soled wading boots when conducting field sampling activities. Neoprene chest waders help to protect against thorns, nettles and poison ivy that may be encountered during the walk to a sampling site, abrasions that may result from instream boulders and logs and from contaminants in the water. Felt-soled wading boots help to prevent slips and falls on slick instream surfaces and also offer added protection when conducting the riffle/run kick during macroinvertebrate sampling.

Field crew members should wear eye protection (safety goggles, glasses) when conducting field sampling activities to protect against branches and splashed water.

A backpack fully loaded with macroinvertebrate sampling equipment (see list in Appendix 10.1) weighs 17 lbs, while the Hydrolab in its backpack weighs 8 lbs. A d-frame dipnet weighs 3 lbs. Therefore, the total weight of equipment carried by one field crew member will usually not exceed 20-22 lbs.

Per recommendations of the IDEM Health and Safety office (IDEM 2010, Dave Appel, personal communication), when heat index temperatures reach 100° F, field work should be reduced to a 50% work /50% rest schedule. Heat index temperatures above 105° F require suspension of field work, until heat index temperatures decrease to below 105° F. Staff collecting the macroinvertebrate sample should exercise caution when elutriating the sample in the bucket as the sample may contain shards of glass, sharp or thorny branches, fish with sharp and/or poisonous spines (such as Brindled Madtom (Noturus miurus)), or macroinvertebrates that can bite (Dobsonflies, Corydalus cornutus), pinch (Crayfish), or pierce the skin (many Hemiptera, notably Giant Water Bugs (Belastoma spp.), Backswimmers (Notonecta spp.), or Water Scorpions (Ranatra spp.)). The forceps used to pick the sample are sharp and may puncture the skin if not handled safely. The 70% ethanol used to preserve the macroinvertebrate sample is flammable and may result in severe eye and moderate skin irritation if mishandled (Ultra Pure 2009, Newhouse 1998a). Use caution during the “pick” portion of the sample, as the sample jar containing 70% ethanol is open at this time. The formalin used to preserve the macroinvertebrate sample is flammable and may result in symptoms ranging from irritation of eyes, skin, and mucous membranes to burning, difficulty breathing, respiratory tract injury, discoloration of skin, roughness and first degree burns if mishandled (Fisher Scientific 1999, Newhouse 1998b). Use caution when adding formalin to the sample container as it may splash onto the hands or face. Field crew adding formalin to the macroinvertebrate sample should wear elbow length rubber gloves and a plastic face shield for added protection.


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5.3.2. Cautions Over time, repeated abrasion against rough objects in the stream will result in the net

portion of the d-frame net developing tears and/or holes. Such tears or holes should be repaired as soon as possible as they will allow collected macroinvertebrates to escape, reducing the number of individual specimens collected and potentially biasing the results. Small tears and holes can be repaired using a strong suturing material (such as fishing line) to sew up the hole. However, numerous or large holes and tears may require the replacement of the entire net.

Sample containers should be filled with 70% ethanol to at least the two-thirds mark. Samples containing a large amount of organic material, snails, freshwater mussels (Corbiculidae, Pisiidae, Dreisennidae), or crayfish will require more 70% ethanol (filled to the top of the container or the use of a second container specifically for those organisms). Failure to adequately preserve the sample may result in degradation of the structural quality of the organisms, hindering their identification.

5.3.3. Interferences Heavy rains of more than one or two inches can often scour the stream substrate,

removing many macroinvertebrates. Sampling a stream soon after a scouring event may result in a sample of poorer quality than what would be normal. A stream should not be sampled less than two weeks after a scouring event to allow for natural macroinvertebrate recolonization by drift.

During the 15 minute pick care should be taken to avoid “scooping” multiple individuals in a single pick. Also, the addition of large pieces of organic material (leaves, algae) should be avoided as they may contain large numbers of individuals and may give skewed results.

See Section 5.3.2. for other possible interferences.

5.3.4. Calibration See Davis 2006 for guidelines on Hydrolab calibration and maintenance.

5.3.5. Troubleshooting See Section 5.3.2. for instructions on repairing holes in the d-frame net.

6.0 Records Management After completion of weekly field sampling activities the sample chain of custody, photo chain of

custody and field notebook entries for the week are scanned into a pdf format. These files are then renamed in the following format: MM-DD-YYYY + crew leaders initials + form type (e.g. Photo Chain of Custody, Field Chain of Custody or Macro Field Notebook) (for example: 07-15-2010_PDM_Photo Chain of Custody). These files are then saved into the following locations: S:\WSP\OWM\RANDOM\Corvallis2010\BSS Chain of Custody Copies\Blue Field Notebook S:\WSP\OWM\RANDOM\Corvallis2010\BSS Chain of Custody Copies\Field Chain of Custody S:\WSP\OWM\RANDOM\Corvallis2010\BSS Chain of Custody Copies\Photo Log Chain of Custody where “Corvalis2010” can be changed to the current year in which field sampling activities are being conducted.

Data entered into the stream data sheet, macroinvertebrate header and QHEI form are entered into AIMS. Original paper copies of all forms are kept on file in the Biological Studies Section of the IDEM Shadeland offices.


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7.0 Quality Assurance / Quality Control Ten percent of sites sampled for macroinvertebrate community are re-sampled for QA/QC

purposes. These duplicate samples are collected by the crew leader during the same sampling event as the original sample. The duplicate sample is collected in an area of the stream that contains habitat similar to that used for the original sample but does not overlap any of the area used for the original sample. The duplicate sample receives separate AA/AB number and macroinvertebrate sample numbers. The purpose of this duplicate sample is to ensure that the sample collector is putting the same effort into the collection and picking of every macroinvertebrate sample that they collect.

It is recommended that if multiple IDEM staff plan to conduct macroinvertebrate sampling, they conduct a series of calibration samples prior to the beginning of the field season. These calibration samples should be collected simultaneously at a site that offers relatively homogenous habitat. Analysis of the number of individuals collected and species diversity should not differ significantly between staff collecting macroinvertebrate samples. Staff from groups outside of the IDEM BSS (other IDEM sections or governmental or non- governmental agencies) should conduct a similar calibration exercise with BSS staff so that they can be properly trained in the application of this sampling method.

8.0 References Aquatic Research Center of the Indiana Biological Survey. 2007.Development of Coolwater Index of Biotic Integrity Expectations for Use in Streams and Rivers of Indiana and Review of Existing Data. Technical Report 2007-01. Indiana Biological Survey, Aquatic Research Center, Bloomington, IN. Davis, T.E. 2006. Biological Studies Section Hydrolab System Callibration, Maintenance and Operation. IDEM Standard Operating Procedure S-001-OWQ-A-BS-06-S-R1. Biological Studies Section, Assessment Branch, Office of Water Management, Indiana Department of Environmental Management, Indianapolis, Indiana. Fisher Scientific. 1999 (revised 2008). Material Safety Data Sheet for Formaldehyde, 37% wt solution, stabilized with methanol. Web address as of September 15, 2010: https://fscimage.fishersci.com/msds/96726.htm IDEM. 1992, revision 1. Section 2, Biological Studies Section Hazards Communications Manual (List of Contents). Biological Studies Section, Surveillance and Standards Branch, Office of Water Management, Indiana Department of Environmental Management, Indianapolis, Indiana. IDEM. 2005. Indiana Department of Environmental Management (IDEM), Office of Water Quality, Assessment Branch, DRAFT: Summary of Protocols: Probability Based Site Assessment for Watershed Program Field Activities, Revision #7, July 28, 2005. IDEM 32/03/002/1999. Indiana Department of Environmental Management, Indianapolis, IN. IDEM. 2007a. Health and Safety Training Policy, A-030-OEA-07-P-R1, revised January 1 2007. Indiana Department of Environmental Management, Office of External Affairs, Indianapolis, Indiana. Web address as of September 15, 2010: http://intranet.idem.in.gov/sites/collaboration/Agency%20Policy/Health%20and%20Safety%20Training%20Policy%20A-030-OEA-07-P-R1.pdf IDEM. 2007b. Injury and Illness Resulting from Occupational Exposure Policy, A-034-OEA-07-P-R1, revised April 25 2007. Indiana Department of Environmental Management, Office of External Affairs, Indianapolis, Indiana. Web address as of September 15, 2010: http://intranet.idem.in.gov/sites/collaboration/Agency%20Policy/Injury%20and%20Illness%20Resulting%20from%20Occupational%20Exposure%20A-034-OEA-07-P-R1.pdf

https://fscimage.fishersci.com/msds/96726.htm�

http://intranet.idem.in.gov/sites/collaboration/Agency%20Policy/Health%20and%20Safety%20Training%20Policy%20A-030-OEA-07-P-R1.pdf�

http://intranet.idem.in.gov/sites/collaboration/Agency%20Policy/Health%20and%20Safety%20Training%20Policy%20A-030-OEA-07-P-R1.pdf�

http://intranet.idem.in.gov/sites/collaboration/Agency%20Policy/Injury%20and%20Illness%20Resulting%20from%20Occupational%20Exposure%20A-034-OEA-07-P-R1.pdf�

http://intranet.idem.in.gov/sites/collaboration/Agency%20Policy/Injury%20and%20Illness%20Resulting%20from%20Occupational%20Exposure%20A-034-OEA-07-P-R1.pdf�


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IDEM. 2008. Personal Protective Equipment Policy, A-059-OEA-08-P-R0, May 1 2008. Indiana Department of Environmental Management, Office of External Affairs, Indianapolis, Indiana. Web address as of September 15, 2010: https://extranet.idem.in.gov/standards/docs/policies/oea/Personal_Protective_Equipment_Policy.pdf IDEM. 2010. Guidance for the prevention of heat stress. Indiana Department of Environmental Management Health and Safety memo dated July 21, 2010. Newhouse, S.A. 1998a. Field and laboratory operating procedures for use, handling and storage of chemicals in the laboratory. IDEM/32/03/007/1998. Biological Studies Section, Assessment Branch, Office of Water Management, Indiana Department of Environmental Management, Indianapolis, Indiana. Newhouse, S.A. 1998b. Field and laboratory operating procedures for use, handling and storage of solutions containing formaldehyde. IDEM/32/03/006/1998. Biological Studies Section, Assessment Branch, Office of Water Management, Indiana Department of Environmental Management, Indianapolis, Indiana. Ohio EPA. 2006. Methods for Assessing Habitat in Flowing Waters: Using the Qualitative Habitat Evaluation Index (QHEI). Ohio EPA Technical Bulletin EAS/2006-06-1. Web address as of September 16, 2010: http://www.epa.state.oh.us/portals/35/documents/QHEIManualJune2006.pdf Sobat, S.L. 2006. Biological Studies Section Qualitative Habitat Evaluation Index (QHEI). IDEM Standard Operating Procedure S-001-OWQ-A-BS-06-S-R1. Biological Studies Section, Assessment Branch, Office of Water Management, Indiana Department of Environmental Management, Indianapolis, Indiana. Ultra Pure. 2009. Material Safety Data Sheet for Reagent Alcohol, 200 Proof. Web address as of September 15, 2010: http://www.ultrapure-usa.com/pages/pdf/msds/reagent_alcohol_200_proof.pdf

9.0 History of Revisions Not applicable because this is the first version of this TSOP.

10.0 Appendices 10.1 Biological Samples Field Chain of Custody form 10.2 Sample field notebook entries 10.3 OWQ Biological Studies: Macroinvertebrate Header form 10.4 OWQ Biological Studies QHEI (Qualitative Habitat Evaluation Index) form (front and back) 10.5 Original Photographic Chain of Custody form 10.6 Stream Sampling Field Data Sheet form 10.7 Site Reconnaissance Aerial Map 10.8 Site Reconnaissance Topographic Map 10.9 Site Reconnaissance Form 10.10 Checklist of equipment for macroinvertebrate sampling

https://extranet.idem.in.gov/standards/docs/policies/oea/Personal_Protective_Equipment_Policy.pdf�

http://www.epa.state.oh.us/portals/35/documents/QHEIManualJune2006.pdf�

http://www.ultrapure-usa.com/pages/pdf/msds/reagent_alcohol_200_proof.pdf�


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Appendix 10.1 Blank Biological Samples Field Chain of Custody form


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Appendix 10.2 Sample field notebook entries


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Appendix 10.3 Blank OWQ Biological Studies: Macroinvertebrate Header form


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Appendix 10.4 Blank OWQ Biological Studies QHEI (Qualitative Habitat Evaluation Index) form (front)


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Appendix 10.4 Blank OWQ Biological Studies QHEI (Qualitative Habitat Evaluation Index) form (back)


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Appendix 10.5 Blank Original Photographic Chain of Custody form


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Appendix 10.6 Blank Stream Sampling Field Data Sheet form


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Appendix 10.7 Sample Site Reconnaissance Aerial Map


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Appendix 10.8 Sample Site Reconnaissance Topographic Map


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Appendix 10.9 Sample Site Reconnaissance Form


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Appendix 10.10 Checklist of equipment for macroinvertebrate sampling (* denotes equipment taken to sampling site) Vehicle binder (containing Wright Express credit card) First-aid kit Fire extinguisher Emergency eye wash 70% ethanol in flame proof safety container Formalin in plastic safety container Face shield Winkler kit Hydrolab maintenance kit Hand sanitizer Bug spray Hydrolab Quanta unit and backpack* External frame backpack* Site file (containing)*: Stream Sampling Field Data Sheet form* OWQ Biological Studies: Macroinvertebrate Header form* OWQ Biological Studies QHEI (Qualitative Habitat Evaluation Index) form* Site Reconnaissance Form and maps* Original Photographic Chain of Custody form* Biological Samples Field Chain of Custody form* Clipboard* Nalgene field notebook* Fine-point Sharpie permanent marker* Lead pencils* 250 ml Nalgene sample containers (3 per site, filled 2/3 full with 70% ethanol)* D-frame dipnet (with extra net and clips)* Two No. 35 Sieves* Two plastic buckets* White plastic tray* Two pairs of forceps* Pelican case (containing)*:

Crown densitometer* Digital camera* GPS unit* Cell phone*

50 meter measuring tape* Neoprene chest waders* Felt-soled wading boots* Hip boots (optional) Personal floatation device (PFD)* Arm length rubber gloves* Safety goggles or glasses*

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