Rapid Bioassessment Protocols: Technical Level

Method 006 – Periphyton Collection

Natural Substrate Single Habitat Sampling

Introduction
(This information is taken from: Stevenson, R.J. and L.L. Bahls. 1999. In: M.T. Barbour, J.
Gerritsen, and B.D. Snyder, eds. Rapid Bioassessment Protocols for use in Wadeable Streams
and Rivers: Periphyton, Benthic Macroinvertebrates, and Fish. Second Edition. EPA 841-B-
99-002 United States Environmental Protection Agency, Washington. Pp 6-1 to 6-22.)

Benthic algae (periphyton) are primary producers and an important foundation of many stream
food webs. These organisms also stabilize substrata and serve as habitat for many other
organisms. Because benthic algal assemblages are attached to substrate, their characteristics are
affected by physical, chemical, and biological disturbances that occur in the stream reach during
the time in which the assemblage developed.

Diatoms in particular are useful ecological indicators because they are found in abundance in
most lotic ecosystems. Diatoms and many other algae can be identified to species by
experienced algologists. The great numbers of species provide multiple, sensitive indicators of
environmental change and the specific conditions of their habitat. Diatom species are
differentially adapted to a wide range of ecological conditions.

As with the macroinvertebrate community, periphyton indices of biotic integrity have been
developed and tested in several regions of the U.S. Periphyton protocols are most effective when
used with one or more of the other assemblages and protocols, especially habitat and benthic
macroinvertebrate assessments. The USEPA Rapid Bioassessment Protocols (Barbour et al.
1999) outlines two sampling methods for wadeable streams: single-habitat and multi-habitat.
Alaska uses the sampling procedures for natural substrates in a single habitat. This provides
more accuracy in assessing biotic integrity and in diagnosing cause of impairment. This type of
sampling should reflect water quality differences among streams more precisely than multi-
habitat sampling, but impacts in other habitats in the reach may be missed. Variability due to
differences in habitat between streams may be reduced by collecting periphyton from a single
substrate/habitat combination that characterizes the study reach (Rosen 1995).

Periphyton samples should be collected during periods of stable stream flow. High flows can
scour the stream bed, flushing the periphyton downstream. Recolonization of substrates will be
faster after less severe floods and in streams with nutrient enrichment. It is recommended that
there is a three-week delay following high, bottom-scouring stream flows to allow for
recolonization and succession to a mature periphyton community (Peterson and Stevenson,
1990). In Alaska, it is recommended that periphyton be collected at the same time as
macroinvertebrate samples and from the same 100 m reach. Within the 100 m reach, four
riffle/cobble subreaches are selected. Within each subreach, four cobbles are selected for

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sampling and a standardized area is sampled from each. In stream reaches lacking appropriate
substrate (cobbles), a standardized area of periphyton can be sampled from bedrock and/or
sediment substrate.

Equipment/Supplies
• Cobble sampler (Rubber periphyton strap)
Bicycle inner tube (largest diameter available, e.g., Mt. bike)
Rubber washer (4.5 cm inside diameter)
Strong, flexible adhesive (e.g., Shoe Goo or Aquaseal)
(Cut a length of inner tube and split it lengthwise to form an approximate 50cm x 10
cm rubber rectangle. In the center of the rectangle, cut a hole slightly larger than the
inside diameter of the rubber washer. Center the rubber washer over this hole and
glue in place.)
• Bedrock sampler
PVC tube (4.5 cm inside diameter x 12 cm long)
Adhesive foam weather stripping (2 cm wide)
Clear packing tape
(Wrap weather stripping around one end of tube so it overhangs the tube end by
about 2mm so it forms a gasket. Wrap weather stripping with packing tape to hold it
in place securely.)
• Sediment sampler
Petri dish (60 x 15 mm)
Kitchen spatula
• Periphyton scrubber
Toothbrush
Epoxy glue
(Cut toothbrush head off and glue perpendicular to the end of the handle. It will look
like a miniature broom.)
• Small bucket
• White washtubs
• Wash bottle (500 mL)
• Spoon or small metal laboratory spatula
• Graduated 1000 mL wide-mouth bottle (Make gradations with a permanent marker)
• Wide-mouth Nalgene sample containers for each site (125 mL)
• Permanent marking pens (Sharpie)
• Preservative (Lugol’s Solution)
• Turkey baster
• Cooler with ice
• Ruler
• Densiometer

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Record Keeping (See page7)
• Field Data Sheet: Periphyton

Field Sampling Procedures

FP-1. Reach selection. Define the sampling reach. (This should be the same 100 m reach that
was sampled for macroinvertebrates.) The recommended substrate/habitat combination is cobble
obtained from riffles and runs with current velocities of 10-50 cm/sec. This habitat is common
in most streams and samples from these areas are easier to analyze because they usually contain
less silt. Identify four distinct riffle/run areas (subreaches) to be sampled. For low gradient
streams with no riffles, sample algae on snags or in run areas.

FP-2. Document site information. Complete the field data sheet: periphyton to identify the
site, date and crew. The habitat and water quality information should already be completed from
the macroinvertebrate sampling.

FP-3. Shade measurement. Measure four forest overstory densities for each subreach using a
densiometer. Measures are taken from the middle of each subreach (1) facing downstream, (2)
facing upstream, (3) facing the left bank, and (4) facing the right bank. Record each reading on
the field data sheet. The measurements are averaged and an overall mean is determined for the
entire reach.

FP-4. Sample collection. Collect 4 cobbles from the downstream subreach and place them in a
single layer in a small, clean bucket containing 1 to 2 cm of water. Take one cobble and while
holding it over a clean, white washtub, wrap the periphyton strap around it and twist the ends of
the strap together to hold it in place. To standardize the area sampled, periphyton will be
sampled only from inside the rubber washer of the periphyton strap. If the periphyton layer is
thick, scrape the periphyton with a spoon or laboratory spatula and rinse into the washtub using a
wash bottle. Continue to alternate scrubbing using the periphyton scrubber and rinsing using the
wash bottle until all periphyton is removed and rinsed into the washtub. This process should
take about 30 seconds. When completed, the area sampled should feel coarse and slime-free.
Repeat this procedure for the other 3 cobbles in the downstream subreach. Continue this process
at the 3 remaining subreaches. In total, 16 cobbles will be sampled from each stream reach (4
cobbles from each of 4 subreaches). All the material collected from the stream reach is
combined to form a single composite sample in the white washtub.

In streams with abundant bedrock, bedrock may be sampled in place of some or all cobbles.
Place the bedrock sampler (gasket-end down) on the bedrock, with the other end protruding out
of the water. Periphyton will only be sampled from inside of the sampler tube to standardize the
sampling area. If the periphyton layer is thick, scrape the bedrock with a spoon or laboratory
spatula. Next, dislodge the periphyton remaining inside the sampler with the periphyton
scrubber. Transfer the material from inside the sampler to the white sample washtub with a
turkey baster. If the inside diameter of the bedrock sampler is different than the inside diameter

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of the periphyton strap, record how many samples are collected with each so the total area
sampled can be adjusted.

In streams where cobble areas are limited, it may be desirable to collect periphyton samples from
streambed sediment. Place a petri dish (open end down) directly on the sediment and push
downward until sediment fills the dish. Slide the spatula beneath the opening of the petri dish to
capture the sediment and associated periphyton. Rinse the contents of the petri dish into the
white sample washtub. Record how many samples are collected with the sediment sampler (vs.
cobbles and/or bedrock) so the total area sampled can be adjusted.

FP-5. Sample preservation. Pour entire sample from the white washtub into the graduated
1000-mL bottle. Record the volume under “initial sample volume” on the Field Data Sheet:
Periphyton. While thoroughly mixing the sample in the graduated bottle, transfer 125 ml of
sample into a 125-ml sample bottle using a clean turkey baster (Bahls 1993). Discard the
remaining sample in the graduated bottle. Add 0.3 ml Lugol’s solution for every 100 ml of
sample. (For example, with a sample volume of 125 ml, add 375 µL [micro liters] of Lugol’s
solution.)

FP-6. Sample labeling. Place a permanent label on the outside of the 125 mL sample container
with the following information: water body name, location, station ID #, date, name of
collector(s), initial sample volume, and type of preservative. Make sure the same information
recorded on the data sheet is recorded on the label. Place another label with the same
information inside the container. (Caution! Lugol’s solution will turn paper labels black.)

FP-7. Biomass assessment. Conduct a rapid survey of moss, macroalgae, and microalgae
biomass growing on stream substrates using the following index. Transect the stream and reach
down and touch a random location on the streambed. Pick up the first particle contacted.
Continue to sample the substrate along at least 3 more transects in this manner until 24 locations
have been sampled in each of the 4 subreaches. (A total of 96 particles will be sampled for each
site when finished). As each particle is selected, measure the size of the particle (in mm) and
record in the Field Data Sheet: Periphyton tally column for the appropriate subreach. For each
particle, characterize the moss, macroalgae, and microalgae biomass using the descriptions in
Table 1. Record these in the tally columns on the field data sheet. Note the same values are used
for moss and macroalgae.

Table 1. Assessment values to index moss, macroalgae, and microalgae biomass.

Periphyton Form Index Value Description
Moss/Macroalgae 0 No macroalgae/moss present
Moss/Macroalgae 1 <5% macroalgae/moss coverage
Moss/Macroalgae 2 5-25% macroalgae/moss coverage
Moss/Macroalgae 3 >25% macroalgae/moss coverage
Microalgae 0 Substrate rough - no apparent growth
Microalgae 0.5 Substrate slimy but no microalgae visible
Microalgae 1 Thin layer of microalgae visible
Microalgae 2 Microalgae 0.5-1 mm in thickness
Microalgae 3 Microalgae 1-5 mm in thickness
Microalgae 4 Microalgae 5-20 mm in thickness
Microalgae 5 Microalgae >2 cm in thickness

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FP-8. Review data sheet. Review all the recorded information on labels, jars, and forms for
accuracy and completeness.

FP-9. Clean tools. Examine all brushing and scraping tools and containers for residues. Rub
them clean and rinse them in distilled water before sampling the next site and before putting
them away.

FP-10. Sample transport. Transport samples back to the laboratory in a cooler with ice (keep
them cold and dark) and store preserved samples in the dark until they are processed. Be sure to
stow samples in a way so that transport and shifting does not cause samples to leak. When
preserved, check preservative every few weeks and replenish as necessary until taxonomic
evaluation is completed.

FP-11. Sample log-in. Upon returning to the laboratory, all incoming samples will be logged in
on the appropriate form (Sample Login/Tracking Sheet: Periphyton). Record sample ID, date
collected, replicate ID if necessary, alternate client sample ID, and comments.

Periphyton Sampling References

Barbour, M.T., J. Gerritsen, B.D.Snyder, and J.B. Stribling. 1999. Rapid Bioassessment
Protocols for Use in Streams and Wadeable Rivers: Periphyton, Benthic Macroinvertebrates and
Fish, Second Edition. EPA 841-B-99-002. U.S. Environmental Protection Agency; Office of
Water; Washington, D.C.

Peterson, C.G. and R.J. Stevenson. 1990. Post-spate development of epilithic algal communities
in different current environments. Canadian Journal of Botany 68:2092-2102.

Rosen, B.H. 1995. Use of periphyton in the development of biocriteria. Pages 209-215 in W.S.
Davis and T.P. Simon (editors). Biological assessment and criteria: Tools for water resource
planning and decision making. Lewis Publishers, Boca Raton, Florida.

Quality Control
QC-1. Labeling. Sample labels must be accurately and thoroughly completed, including the
sample identification code, date, stream name, sampling location, and collector’s name. The
outside and any inside labels of the container should contain the same information. Sample log
forms must include the same information as the sample container labels. Caution! Lugol’s
solution and iodine-based preservatives will turn paper labels black.

QC-2. Clean equipment. After sampling has been completed at a given site, all brushes,
suction and scraping devices that have come in contact with the sample should be rubbed clean
and rinsed thoroughly in distilled water. The equipment should be examined again prior to use at
the next sampling site and rinsed again if necessary.

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QC-3. Review information. After sampling, review the recorded information on all labels and
forms for accuracy and completeness.

QC-4. Replicate sample. Collect and analyze one replicate sample from 10% of the sites to
evaluate precision or repeatability of sampling technique, collection team, sample analysis, and
taxonomy.

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FIELD DATA SHEET: Periphyton
Densiometer Measures: Percent Shade
Site ID_________________ Sub Sub Sub Sub
Date:__________________ Reach 1 Reach 2 Reach 3 Reach 4
Right Bank
Site Name______________________ Upstream
Crew Initials _______________ ____ Left Bank
Initial Sample Volume____________ Downstream
Average

Biomass Assessment

Subreach 1 Subreach 2 Subreach 3 Subreach 4

Algal Biomass Class Tallies Total Tallies Total Tallies Total Tallies Total
Not suitable substrate

Moss 0
Moss 1
Moss 2
Moss 3
Macroalgae 0
Macroalgae 1
Macroalgae 2
Macroalgae 3

Microalgae 0
Microalgae 0.5
Microalgae 1
Microalgae 2
Microalgae 3
Microalgae 4
Microalgae 5
Number of Suitable Substrates Counted: Median Moss Cover:

Median Macroalgal Cover: √ if macroalgal sample collected Median Microalgal Cover:

Stream Bed Particle Subreach 1 Subreach 2 Subreach 3 Subreach 4

Size Counts
Tallies Total Tallies Total Tallies Total Tallies Total

Bedrock
Boulder >256mm
Cobble 160-256mm
Cobble 64-160mm
Gravel 33-64mm
Gravel 2-33mm
Sand .06-2mm
Silt <.06mm
Total # of particles counted: Median:

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 Rapid Bioassessment Protocols: Technical Level
Method 007 – Periphyton Processing
(This information is from: Stevenson, R.J. and L.L. Bahls. 1999. In: M.T. Barbour, J.
Gerritsen, and B.D. Snyder, eds. Rapid Bioassessment Protocols for use in Wadeable Streams
and Rivers: Periphyton, Benthic Macroinvertebrates, and Fish. Second Edition. EPA 841-B-
99-002 United States Environmental Protection Agency, Washington. Pp 6-1 to 6-22.)

At this time it is recommended that only diatoms be identified. In general, identifying diatoms to
species is recommended for two reasons: (1) to better characterize differences between
assemblages that may occur at the species level and (2) because large differences in ecological
preferences do exist among algal species within the same genus. However, substantial
information can be gained by identifying diatoms just to the genus level. Whereas identifying
diatoms only to genus may loose valuable ecological information, costs of analyses can be
reduced, especially for inexperienced analysts. If implementing a new program and only an
inexperienced analyst is available for the job, identifying diatom genera in assemblages can
provide valuable characterizations of biotic integrity and environmental conditions. For an
experienced taxonomist, the cost of identifying diatoms to species is not much greater than
identifying to genus.

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Equipment/Supplies
Caution! (Fume hood and protective clothing required for processing.)
• Fume Hood
• Graduated beakers (1-400 mL for each sample)
• Tissue homogenizer (hand-held blender)
• Adjustable pipette (50 – 1000 µL)
• Pipette tips to fit adjustable pipette (one for each sample)
• 5000 µL pipette
• 5000 µL pipette tips (one tip for each sample)
• Sample storage vials (6 dram glass with screw top – 2 for each sample)
• Stir plate
• Magnetic stirring rod
• Stirring rod retriever
• Hot plate (in hood)
• Wash bottle
• Deionized water
• Tray
• Laboratory coat, safety goggles, and gloves
• Vacuum hose (with fine tip)
• Forceps
• Chemical spatula
• Microscope slides
• Cover slips (22 mm x 22 mm glass)
• Ceramic tiles
• Aluminum foil
• Straight razor blade
• pH paper
• Nitric acid (concentrated)
• Potassium dichromate
• Naphrax (specialty item)

Record Keeping (See pages 14 and 15 )

• Sample Login/Tracking Sheet: Periphyton
• Laboratory Processing Data Sheet: Periphyton

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Sample Preparation
SP-1. Sample identification. Record the sample identification number on the Login/Tracking
sheet and also on the Laboratory Processing Data Sheet.

SP-2. Beaker labeling. Label one beaker for each sample with the sample identification
number.

SP-3. Vial labeling. Label 2 sample storage vials with the sample identification number for
each sample. Write “Diatom” on one vial and “Soft” on the other vial.

SP-4. Set up. Organize samples and beakers to facilitate efficient processing.

Sample Homogenization
SH-1. Sample volume. Record the initial sample volume on the laboratory processing sheet.

SH-2. Pour. Pour contents of the sample bottle into the graduated beaker. Thoroughly rinse the
bottle into the beaker with deionized water.

SH-3. Homogenize. Homogenize the algal sample with a tissue homogenizer or hand blender
until thoroughly mixed. Using a wash bottle, rinse the homogenizer with deionized water into
the beaker and rinse down the walls of the beaker. Bring the volume up to the next marked
volume on the beaker.

SH-4. Record. Record the new volume in the “working volume” column on the laboratory
processing data sheet

SH-5. Stir. Insert the stir bar and stir sample. Remember to rinse the stir bar with deionized
water before and after each use.

SH-6. Subsample. Take a subsample from the working volume using the 5000-µL pipette with
a clean tip. Subsamples should be taken only when the sample is stirring and from the middle of
the beaker, not from the top). If the sample is very silty, it is better to subsample for soft algae a
minimum of 10 mL. If the sample is clear, sample at least 20 mL. In general, sample at least 10
mL for soft and at least 20 mL for diatoms.
a. Diatom split: used for acid digested clean counts
Place into labeled beaker for acid digestion. Record the volume on the laboratory
processing sheet. Usually, this will be a volume of 10 mL. If the sample is
exceptionally sparse or silty, use a larger volume.
b. Soft split: used for soft algae counts
Place into a 25 mL vial labeled with the sample identifier. Record the volume on the
laboratory processing sheet. Generally, this will be 20 mL for the soft split but it may

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 be reduced if a large volume was used for the diatom split. Set this vial aside until
sample identification.

SH-7. Place the rest of the sample (remnant) back into the original sample bottle. If the volume
exceeds the limits of the bottle, let the sample settle for at least 9 hours and siphon off the
necessary supernatant so the remainder of the sample will fit into the bottle.

Sample Digestion
SD-1. Preparation. Set the hot plate dial to 200° C. Put on laboratory coat, gloves, and
goggles. Place tray containing all subsample beakers under the fume hood.

SD-2. Nitric acid addition. Add enough nitric acid to each beaker to increase the volume to 50
mL. (As a rule, the minimum should be a 1:2 sample to acid ratio.)

SD-3. Heat. Transfer the beakers to the hotplate and heat for 2 hours. Pay careful attention to
samples while heating. If the volume drops too low, add deionized water. When finished
heating, add a dash of potassium dichromate to each beaker to catalyze the reaction. Keep
adding small amounts until further reactions cease.
SD-4. Cool and dilute. After the beakers have cooled somewhat, transfer them back to the tray.
Top off the bakers with deionized water and allow diatom frustules to settle for 12 hours.

SD-5. Siphon and add water. Using a fine-tipped vacuum hose, draw down the samples to
approximately 50 mL. Siphon the water from the center of the water column under the surface.
Make sure not to siphon the diatom layer off the bottom of the beaker. After siphoning, add
deionized water to replace the supernatant drawn off and wash the sides of the beaker to remove
diatoms adsorbed to the sides and top of the beaker. Let settle again for at least 9 hours. (As a
rule, let settle 1 hour per 1 centimeter so the smallest diatoms can settle out.)

SD-6. Repeat step SD-5. Repeat siphoning and addition step another 5 or 6 times and test the
pH with pH paper. When the samples are approximately neutral, the samples are ready for slide
mounting.

SD-7. Reduce volume. Draw the sample volumes down to between 25-50 mL, making sure not
to remove diatoms.

SD-8. Transfer and record volume. Transfer the remaining volume to labeled vials and record
diatom volume after digestion on the laboratory processing sheet. Make sure to rinse all diatoms
clinging to the beaker into the sample vial with deionized water. If the full volume does not fit
into the vial, allow the vial contents to settle for at least 12 hours and siphon off some of the
supernatant. Transfer the remaining contents of the beaker into the vial, again making sure to
rinse all remaining diatoms into the sample vial.

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Slide Preparation

tile
cover slip
labeling tape

SP-1. Set up. Lay out your set-up. Place cover slips on clean, dry ceramic tiles. Place tape
down and label with sample name and the volume of sample to be placed on the cover slip.

SP-2. Add water. Using a clean pipette tip, carefully place an appropriate amount of deionized
water on the cover slip so that when the sample material is added (see SP-3) the volume should
total approximately 1.25 mL.

SP-3. Add sample and mix. Shake the sample vial very well and add sample to the water on
the cover slip. The amount of sample needed will vary based on the diatom concentration. It is
better to add too little sample than too much. With experience it is possible to estimate the
amount needed, but start with 200 µL or less. Record this volume in the “Drip cover slip”
column of the Laboratory Processing Data Sheet. Make sure that the mixture reaches to the
corners of the cover slip. Adjust the pipette to 0.3 mL or more and draw the solution up into the
pipette and back out a few times to evenly distribute the mixture on the cover slip.

SP-4. Cover and dry. When finished with the “row”, loosely cover the tiles with aluminum foil
(make sure the foil does not touch the cover slips). Allow the cover slips to dry.

SP-5. Check and evaluate. When cover slips are dry, check them under the microscope at
400X total magnification. Make sure that there are ~20 to 25 diatom valves in your field of
view. Also make sure that the cells are visible (i.e., that there is not too much sand and silt).
a. If the cover slip is too dense, start over.
b. If it is not dense enough, add more material to it (follow the same steps as before).
Record this volume in the “Drip additional” column of the Laboratory Processing
Data Sheet.
c. If the density is about right, proceed to fix the cover slip to a slide.

SP-6. Fix cover slip to slide. Label a clean slide (site code and date) and place about 2-3 drops
of NAPHRAX on the center of the slide using a small wooden stick. Carefully, pick up the
cover slip with forceps and place it inverted (material side down) on the NAPHRAX.

SP-7. Heat slide. Under a hood, place slide on hotplate (start with medium-low setting and
adjust as necessary). Boil the NAPHRAX (solvent) off. (The solvent contains toluene so this
must be done under a hood.) Allow the solvent to thin out and reach the edges. With the
forceps, make sure that the cover slip is in the center of the slide and the NAPHRAX has reached
all the edges. Allow slide to warm until bubbling slows down (about 30 seconds.)

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SP-8. Cool. Pull slide off the warmer. Use the forceps to position the cover slip squarely in the
middle of the slide. Set aside to cool. When the slide has cooled, remove the excess resin with a
straight razor blade.

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Sample Login/Tracking Sheet: Periphyton

Sample ID Date Replicate ID Client Alt. ID Comments

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 Laboratory Processing Data Sheet: Periphyton (record volume in mL)
Sample ID Initial Working Diatom Soft Diatom DRIP DRIP Comments Date
Sample Vol from SPLIT Algae Vol after cover Additi
Vol split SPLIT digestion slip onal

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Rapid Bioassessment Protocols: Technical Level
Method 008 – Periphyton Identification

Equipment/Supplies
• Microscope (1000X, oil immersion lens, ocular micrometer calibrated with stage micrometer)
• Immersion oil
• Lens paper
• Kimwipes
• Palmer counting cell
• 100 µL pipette and tips
• Taxonomic keys

Record Keeping (See pages 18 and 19)

• Laboratory Bench Sheet –Periphyton Taxonomic Identification
• Laboratory Bench Sheet – Diatom Live/Dead Count

Identification
ID-1. Record information. Record the station identification code, stream name, collection
date, sample collector initials, taxonomist initials, and identification date on the identification
bench sheet.

ID-2. Prepare soft algae sample for live/dead count. Thoroughly homogenize the soft algae
sample and pipette several drops into a Palmer counting cell. Add more of the sample or dilute
as necessary to achieve an algal density of 10 to 20 cells within the field of view (400X). This
density will maximize counting speed for the live/dead count.

ID-3. Count and record live/dead count. Record tallies of both live and dead diatom valves in
the tally column on bench sheet until 300 valves have been counted. Be sure to count 2 valves
for each intact frustule. While counting, move the counting cell in a systematic fashion to avoid
re-counting diatoms.

ID-4. Count and identify diatoms. Use 1000X magnification under oil immersion to count
and identify diatom valves in the acid-digested diatom slide. Keep a tally on the Laboratory
Bench Sheet- Diatom Identification. Diatoms should be identified to the lowest taxonomic level
practical, species if possible, using the references listed. Move the slide systematically to avoid
re-counting valves. Continue counting until 600 valves have been identified (count both valves
on intact frustules) and at least 10 valves of 10 species have been identified.

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Quality Control
QC-1. Samples will be split and duplicate slides sent to periphyton taxonomists for
identification and to verify results.

Periphyton Taxonomic References

Krammer, K. and H. Lange-Bertalot. 1986 – 1991. Susswasserflora von Mitteleuropa. Band 2.
Parts 1-4. Bacillariophyceae. Gustav Fischer Verlag. Stuttgart. New York.

Patrick, R. and L.R. Freese. 1961. Diatoms (Bacillariophyceae) from Northern Alaska.
Proceedings of the Academy of Natural Sciences of Philadelphia 112:129-293.

Patrick, R. and C.W. Reimer. 1975. The Diatoms of the United States. Vol. 2, Part 1.
Monograph No. 13. Academy of Natural Sciences, Philadelphia, Pennsylvania.

Prescott, G.W. 1962. The algae of the Western Great Lakes area. Wm. C. Brown Co.,
Dubuque, Iowa.

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Laboratory Bench Sheet: Diatom Identification
Station ID_______________ Stream_________________ Date_____________
Collected by____________ Slide Prep by____________
Subsample target_________ Taxonomy by____________ ID Date__________

Taxa Tally (No. of Valves) Total Valves TCR*

*Taxonomic Certainty Rating: 5=most certain, 1=least certain. For rating of 1-3, give reason.

Total No. Valves: ___________ Total No. Taxa: ___________

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 Laboratory Bench Sheet: Diatom Live/Dead Count

Station ID________________ Stream____________________ Date_____________

Collected by_______________ Slide Prep by_______________
Subsample target__________ Taxonomy by_______________ ID Date__________

Live Tally (No. of Valves) Dead Tally (No. of Valves)

NOTES:
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Periphyton Assessment Methods 006 to 008 • 1st Edition • ENRI • Page 19 of 19