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8.d) Zavorals Creek September 2013 Technical Summary Macroinvertebrae-WCDAnalysis of a biological sample:
Technical summary of methods and quality assurance procedures
Prepared for Washington County Conservation District
Erik Anderson, Project Manager
December 13, 2013
by
W. Bollman, Chief Biologist
Rhithron Associates, Inc.
Missoula, Montana
METHODS
Sample processing
One macroinvertebrate sample collected from Zavoral's Creek was delivered to Rhithron's
laboratory facility in Missoula, Montana on September 20, 2013. The sample arrived in good
condition. The sample was decanted prior to shipment; preservative was replenished immediately
upon arrival at the laboratory. A chain of custody document containing sample identification
information was provided by the Washington County Conservation District (WCD) Project
Manager. Upon arrival, the sample was unpacked and examined, and checked against the WCD
chain of custody. An inventory spreadsheet including project code and internal laboratory
identification numbers was uploaded into the Rhithron database prior to sample processing.
A subsample of a minimum of 300 organisms was achieved using methods consistent
with Minnesota Pollution Control Agency protocols (MPCA 2004). A Caton sub -sampling device
(Caton 1991), divided into 30 grids, each approximately 6 cm by 6 cm was used. The sample was
thoroughly mixed in its jar, poured out and evenly spread into the Caton tray, and individual grids
were randomly selected. The contents of each grid were examined under stereoscopic
microscopes using 10x -30x magnification. Grid selection, examination, and sorting continued until
at least 300 organisms were sorted. All aquatic invertebrates from each selected grid were sorted
from the substrate, and placed in 80% ethanol for subsequent identification.
After the target number of organisms was obtained in the subsample, a large/rare search
was performed: the Caton tray was scanned for additional organisms that were not collected in
the subsample. These organisms were placed in a separate vial and labeled as "Large/Rare
Organisms". Unsorted sample fractions were retained and stored at the Rhithron laboratory.
Organisms were individually examined by certified taxonomists, using 10x — 80x
stereoscopic dissecting scopes (Leica S8E) and identified to target taxonomic levels consistent
with MPCA stream protocols (MPCA 2004), using appropriate published taxonomic references and
keys.
Identification, counts, life stages, and information about the condition of specimens were
recorded on electronic bench sheets. Organisms that could not be identified to the taxonomic
targets because of immaturity, poor condition, or lack of complete current regionally -applicable
published keys were left at appropriate taxonomic levels that were coarser than those specified.
To obtain accuracy in richness measures, these organisms were designated as 'snot unique" if
other specimens from the same group could be taken to target levels. Organisms designated as
"unique" were those that could be definitively distinguished from other organisms in the sample.
Taxonomists examined organisms in the Large/Rare vial and determined there were not any
unique taxa collected. Identified organisms were preserved in 80% ethanol in labeled vials, and
archived at the Rhithron laboratory.
Chironomids were carefully morphotyped using IN — 80x stereoscopic dissecting
microscopes (Leica S8E) and representative specimens were slide mounted and examined at
200x — 1000x magnification using an Olympus BX 51 or Leica DM 1000 compound microscope.
Slide mounted organisms were archived at the Rhithron laboratory.
Quality control procedures
Quality control procedures for initial sample processing and subsampling involved
checking sorting efficiency. An independent observer microscopically re-examined 25% of sorted
substrate from the sample. Quality control procedures proceeded as follows:
The quality control technician poured the sorted substrate from the processed sample
out into a Caton tray, redistributing the substrate so that 25% of it could be accurately lifted out
by removing entire grids in a random fashion. Grids were selected, and re-examined until 25% of
the substrate was re -sorted. All organisms that were missed were counted and this number was
added to the total number obtained in the original sort. Sorting efficiency was evaluated by
applying the following calculation:
SE = n, x 100
n, +n2
where: SE is the sorting efficiency, expressed as a percentage, nl is the total number of
specimens in the first sort, and nZ is the total number of specimens expected in the second sort,
based on the results of the re -sorted 25%.
Quality control procedures for taxonomic determinations of invertebrates involved
checking accuracy, precision and enumeration. All organisms were re-identified and counted by
an independent taxonomist. Taxa lists and enumerations were compared by calculating a Bray -
Curtis similarity statistic (Bray and Curtis 1957), Percent Taxonomic Disagreement (PTD) and
Percent Difference in Enumeration (PDE). Routinely, discrepancies between the original
identifications and the QC identifications are discussed among the taxonomists, and necessary
rectifications to the data are made. Discrepancies that cannot be rectified by discussions are
routinely sent out to taxonomic specialists for identification. However, taxonomic certainty for
identifications in this project was high and no external verifications were necessary.
Data analysis
Taxonomic data and counts were entered into the Rhithron laboratory database, and an
uploadable spreadsheet file, consistent with MPCA stream data requirements, including
taxonomy, counts, life stages, sample metadata, and other information was generated.
Taxa lists and counts were constructed and standard metric calculations for aquatic
invertebrate assemblages were made using Rhithron's customized database software. A sites -by -
taxa matrix in spreadsheet format was also created.
RESULTS
Quality Control Procedures
Sorting efficiency was 95.40%, taxonomic precision for identification and enumeration
was 98.01% (Bray Curtis), 0.93% PTD and 1.07% PDE, and data entry efficiency was 100%.
These similarity statistics fall within acceptable industry criteria (Stribling et al. 2003).
Data analysis
Taxa list and metric summary pages are given in the Appendix. Electronic spreadsheets
were provided to the WCD Project Manager via email.
2
REFERENCES
Bray, J. R. and J. T. Curtis. 1957. An ordination of upland forest communities of southern
Wisconsin. Ecological Monographs 27: 325-349.
Caton, L. W. 1991. Improving subsampling methods for the EPA's "Rapid Bioassessment" benthic
protocols. Bulletin of the North American Benthological Society. 8(3): 317-319.
MPCA, 2004. Laboratory Analysis — Invertebrate Sample Processing and Identification.
State of Minnesota Standard Operating Procedure.
Stribling, J.B., S.R Moulton II and G.T. Lester. 2003. Determining the quality of taxonomic data.
J.N. Am. Benthol. Soc. 22(4): 621-631.
3
APPENDIX
Taxa list and metric summary
Washington County Conservation District, MN
Zavoral's Creek
September 2013
Taxa Listing
Yes
RAI No.: WCD13ZC2001
5
Client ID:
0.31%
Date Coll.: 9/17/2013
No. Jars: 1
Taxonomic Name
Count
Other Non -Insect
Yes
Acari
11
Nemata
1
Turbellaria
2
Asellidae
CG
Caecidotea sp.
3
Gammaridae
4
Gammarus sp.
69
Physidae
Unknown
Physa sp.
22
Oligochaeta
Yes
Oligochaeta
1
Ephemeroptera
12.62%
Baetidae
Larva
Baetis sp.
41
Leptophlebiidae
Yes
Paraleptophlebia sp.
2
Plecoptera
4.92%
Capniidae
Larva Early Instar
Capniidae
16
Nemouridae
Yes
Amphinemura sp.
1
Trichoptera
0.31%
Glossosomatidae
Larva
Glossosoma sp.
1
Hydropsychidae
Yes
Diplectrona modesta
2
Hydropsychidae
6
Lepidostomatidae
Larva Early Instar
Lepidostoma sp.
2
Limnephilidae
Yes
Hesperophylax designatus
2
Limnephilus sp.
3
Psychomyiidae
Larva
Lype diversa
4
Rhyacophilidae
Yes
Rhyacophila sp.
9
Friday, December 13, 2013
Project ID: WCD13ZC2
RAI No.: WCD13ZC2001
Sta. Name: Zavoral's Creek
STORET ID:
PRA Unique Stage Qualifier BI Function
3.38%
Yes
Unknown
5
PR
0.31%
Yes
Unknown
5
UN
0.62%
Yes
Unknown
4
PR
0.92%
Yes
Unknown
8
CG
21.23%
Yes
Unknown
4
SH
6.77%
Yes
Unknown
8
SC
0.31%
Yes
Unknown
10
CG
12.62%
Yes
Larva
5
CG
0.62%
Yes
Larva
1
CG
4.92%
Yes
Larva Early Instar
1
SH
0.31%
Yes
Larva
2
SH
0.31%
Yes
Larva
0
SC
0.62%
Yes
Larva
0
CF
1.85%
No
Larva Early Instar
4
CF
0.62%
Yes
Larva
1
SH
0.62%
Yes
Larva
5
SH
0.92%
Yes
Larva
3
SH
1.23%
Yes
Larva
2
SC
2.77%
Yes
Larva
1
PR
Taxa Listing
STORET ID:
RAI No.: WCD13ZC2001
Client ID:
PRA
Date Coll.: 9/17/2013 No. Jars:
1
Taxonomic Name
Count
Diptera
0.31%
Ceratopogonidae
Larva
Ceratopogoninae
1
Dixidae
2.46%
Dixa sp.
8
Empididae
1
Empididae
1
Neoplasta sp.
1
Ptychopteridae
6
Ptychoptera sp.
1
Simuliidae
Larva
Simulium sp.
17
Chironomidae
0.31%
Chironomidae
Larva
Corynoneura sp.
2
Eukiefferiella sp.
20
Heterotrissocladius sp.
1
Heterotrissocladius sp.
1
Micropsectra sp.
4
Odontomesa sp.
3
Orthocladiinae
2
Parametriocnemus sp.
1
Parametriocnemus sp.
16
Paratendipes sp.
1
Polypedilum sp.
2
Prodiamesa sp.
3
Synorthocladius sp.
2
Tanytarsus sp.
1
Tanytarsus sp.
1
Thienemanniella sp.
2
Thienemannimyia Gr.
1
Tvetenia sp.
34
Zavrelimyia sp.
1
Sample Count
325
Friday, December 13, 2013
Project ID: WCD13ZC2
RAI No.: WCD13ZC2001
Sta. Name: Zavoral's Creek
STORET ID:
PRA
Unique
Stage
Qualifier
131
Function
0.31%
Yes
Larva
Early Instar
6
PR
2.46%
Yes
Larva
1
CG
0,31%
No
Larva
Early Instar
6
PR
0.31%
Yes
Larva
5
PR
0.31%
Yes
Larva
7
CG
5.23%
Yes
Larva
6
CF
0.62%
Yes
Larva
7
CG
6.15%
Yes
Larva
8
CG
0.31%
No
Pupa
0
CG
0.31%
Yes
Larva
0
CG
1.23%
Yes
Larva
4
CG
0.92%
Yes
Larva
4
CG
0.62%
No
Pupa
Damaged
6
CG
0.31%
No
Pupa
5
CG
4.92%
Yes
Larva
5
CG
0.31%
Yes
Larva
10
CG
0.62%
Yes
Larva
6
SH
0.92%
Yes
Larva
3
CG
0.62%
Yes
Larva
2
CG
0.31%
No
Pupa
6
CF
0.31%
Yes
Larva
6
CF
0.62%
Yes
Larva
6
CG
0.31%
Yes
Larva
5
PR
10.46%
Yes
Larva
5
CG
0.31%
Yes
Larva
8
PR
Project ID: WCD13ZC2
RAI No.: WCD13ZC2001
Ste. Name: Zavoral's Creek
Client ID:
STORET ID
Coll. Date: 9/17/2013
Latitude:
Abundance Measure:
Sample Count:
Sample Abundance:
Coll. Procedure:
Sample Notes:
Longitude:
325
1,950.00 16.67% of sample used
Taxonomic Composition
Category
R
Terrestrial
7
Other Non -Insect
6
Oligochaeta
1
Odonata
3
Ephemeroptera
2
Plecoptera
2
Heteroptera
MTV
Meqaloptera
3
Neuroptera
7
Trichoptera
7
Lepidoptera
1
Coleoptera
Diptera
5
Chironomidae
15
Dominant Taxa
44.31
Category
78.77%
Gammarus
■ Lepi dopter a
Beets
692
Tvetenia
4.010
Phvsa
®Diptera
Eukiefferiella
0.096
Simulium
29
Parametriocnemus
Im
Capniidae
30.15%
Acari
8.31
Rhvacophila
3
Dixa
8.31%
Hvdroosvchidae
53.85%
Micropsectra
37.54%
Lvpe diversa
1.000
Caecidotea
0.500
Functional Composition
Category
R
Predator
7
Parasite
E Ter r estr i al
Collector Gatherer
17
Collector Filterer
3
Macrophvte Herbivore
■Dl i gochaeta
Piercer Herbivore
27
Xvlophaqe
MTV
Scraper
3
Shredder
7
Omnivore
17
Unknown
1
A
PRA
E Richness
Biolndex
Description
E Ter r estr i al
108
33.23%
■ Other No.-mee.t
1
0.31%
■Dl i gochaeta
Montana DEQ Plains (Bukantis 1998)
27
■ Dd..aie
MTV
Montana Revised Valleys/Foothills (Bollman 1998)
0 Eph-, pt.ra
43
13.23%
DPle.optera
17
5.23%
0Heter.Pt
Dominance
0 M egal ptar a
21.23%
DominantTaxa (2) Percent
Neu, opter e
Dominant Taxa (3) Percent
44.31
T, i chapter a
78.77%
Diversity
■ Lepi dopter a
29
692
Col eoptera
4.010
Marqalef D
®Diptera
Simpson D
0.096
D Chi r on..i da.
29
8.92%
Im
98
30.15%
Predator Percent
A PRA
69 21.23%
41 12.62%
34 10.46%
22 6.77%
20 6.15%
17 5.23%
17 5.23%
16 4.92%
11 3.38%
9 2.77%
8 2.46
6 1.85%
4 1.23%
4 1.23%
3 0.92%
A PRA
27 8.31
0 Cal Iec rFi I terer
148 45.54% 0 Col l ector Gatherer
M..r.phyte Herbiv re
27 8.31% 11110-w.re
1111P. -in.
OPier- Herbivore
E P r edamr
0s raper
27 8.31%
■ shredder
95 29.23% ■Unkn.wn 7)
0 Xyl .phage
1 0.31
Bioassessment Indices
38
E Richness
Biolndex
Description
Score
Pct Rating
BIBI
B -IBI (Karr at al.)
20
40.00%
MTP
Montana DEQ Plains (Bukantis 1998)
27
90.00% None
MTV
Montana Revised Valleys/Foothills (Bollman 1998)
10
55.56% Slight
MTM
Montana DEQ Mountains (Bukantis 1998)
11
52.38% Moderate
Friday, December 13, 2013
Metric Values and Scores
Metric Value
Composition
Taxa Richness
38
E Richness
2
P Richness
2
T Richness
7
EPT Richness
11
EPT Percent
27.38%
All Non -Insect Abundance
109
All Non -Insect Richness
7
All Non -Insect Percent
33.54%
Oligochaeta+Hirudinea Percent
0.31%
Baetidae/Ephemeroptera
0.953
Hvdropsvchidae/Trichoptera
0.276
Dominance
Dominant Taxon Percent
21.23%
DominantTaxa (2) Percent
33.85%
Dominant Taxa (3) Percent
44.31
Dominant Taxa It 0) Percent
78.77%
Diversity
Shannon H (loqe)
2.779
Shannon H (log2)
4.010
Marqalef D
6.439
Simpson D
0.096
Evenness
0.054
Function
Predator Richness
7
Predator Percent
8.31
Filterer Richness
3
Filterer Percent
8.31%
Collector Percent
53.85%
Scraper+Shredder Percent
37.54%
Scraper/Filterer
1.000
Scraper/Scraper+Filterer
0.500
Habit
Burrower Richness
3
Burrower Percent
0.92
Swimmer Richness
3
Swimmer Percent
15.69%
Clinqer Richness
6
Clinqer Percent
16.00
Characteristics
Cold Stenotherm Richness
0
Cold Stenotherm Percent
0.00%
Hemoqlobin Bearer Richness
2
Hemoqlobin Bearer Percent
0.92%
Air Breather Richness
1
Air Breather Percent
0.31%
Voltinism
Univoltine Richness
16
Semivoltine Richness
0
Multivoltine Percent
47.08%
Tolerance
Sediment Tolerant Richness
1
Sediment Tolerant Percent
0.31
Sediment Sensitive Richness
1
Sediment Sensitive Percent
0.31
Metals Tolerance Index
3.316
Pollution Sensitive Richness
1
Pollution Tolerant Percent
30.46%
Hilsenhoff Biotic Index
4.665
Intolerant Percent
15.08%
Supertolerant Percent
14.77%
CTQa
91.147
100%
80%
60%
40%
20%
0%
BIBI MTM MTP MTV
Bioassessment Indices