National Rivers and Streams Assessment 2013/14           Field Operations Manual
Version 1.0, May 2013                          Wadeable
             United States Environmental Protection Agency
             Office of Water
             Office of Environmental Information
             Washington, DC
             EPA-841-B-12-009b
     National Rivers and Streams
         Assessment 2013/14
      Field Operations
             Manual
           Wadeable
May 2013

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National Rivers and Streams Assessment 2013/14                            Field Operations Manual
Version 1.0, May 2013                                                               Wadeable

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National Rivers and Streams Assessment 2013/14                          Field Operations Manual
Draft Version 1.0, May 2013                                                      Wadeable

NOTICE


The complete documentation of overall NRSA project management, design, methods, and
standards is contained in four companion documents, including:
National Rivers and Streams Assessment: Quality Assurance Project Plan EPA-841-B-12-007
National Rivers and Streams Assessment: Site Evaluation Guidelines EPA-841-B-12-008
National Rivers and Streams Assessment: Field Operations Manual EPA-841-B-12-009 a and b
National Rivers and Streams Assessment: Laboratory Methods Manual EPA 841-B-12-010
This document (Field Operations Manual (FOM)) contains a brief introduction and procedures to
follow at the base location and on-site, including methods for sampling water chemistry (grabs
and in situ measurements), periphyton, benthic macroinvertebrates, microcystins, fish
assemblage, fish tissue plugs, whole fish tissue, Enterococci, and physical habitat. These
methods are based on the guidelines developed and followed in the National Rivers and Streams
Assessment 2008-2009 (EPA 2012), Western Environmental Monitoring and Assessment
Program (Baker, et al., 1997), the methods outlined in Concepts and Approaches for the
Bioassessment of Non-wadeable Streams and Rivers (Flotemersch, et al., 2006), and methods
employed  by several key states that were involved in the planning phase of this project.
Methods described in this document are to be used specifically in work relating to the National
Rivers and Streams Assessment 2013/14. All Project Cooperators must follow these guidelines.
Mention of trade names or commercial products in this document does not constitute
endorsement or recommendation for use. Details on specific methods for site evaluation and
sample processing can be found in the appropriate companion document.
The suggested citation for this document is:
USEPA. 2013. National Rivers and Streams Assessment 2013-2014: Field Operations Manual -
Wadeable. EPA-841-B-12-009b. U.S. Environmental Protection Agency, Office of Water
Washington, DC.

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         National Rivers and Streams Assessment 2013/14                            Field Operations Manual
         Version 1.0, May 2013                                                               Wadeable
IV

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National Rivers and Streams Assessment 2013/14                            Field Operations Manual
Draft Version 1.0, May 2013                                                           Wadeable


TABLE OF CONTENTS

NOTICE	Ill

TABLE OF CONTENTS	V
LIST OF FIGURES	VIM

LIST OF TABLES	IX

ACRONYMS/ABBREVIATIONS	XI

DISTRIBUTION LIST	XIII

1    BACKGROUND	1

   1.1    SURVEY DESIGN	1
   1.2    TARGET POPULATION AND INDEX PERIOD	1
   1.3    REPLACING SITES	2
   1.4    SELECTION OF NRSA INDICATORS	2
   1.5    SUPPLEMENTAL MATERIAL TO THE FIELD OPERATIONS MANUAL	3
   1.6    RECORDING DATA AND OTHER INFORMATION	5

2    INTRODUCTION TO WADEABLE SAMPLING	8

   2.1    DAILY OPERATIONS	8
   2.2    BASE SITE ACTIVITIES	10
     2.2.1   Pre-departure Activities	10
     2.2.2   Post Sampling Activities	12
   2.3    SAFETY AND HEALTH	14
     2.3.1   General Considerations	14
     2.3.2   Safety Equipment	16
     2.3.3   Safety Guidelines for Field Operations	16
   2.4    FORMS (PAPER OR ELECTRONIC)	18
     2.4.1   Field Forms	18
     2.4.2   Tracking Forms	18
     2.4.3   Equipment and Supplies	19

3    INITIAL SITE PROCEDURES	20

   3.1    SITE VERIFICATION ACTIVITIES	20
     3.1.1   Locating the X-Site	20
     3.1.2   Determining the Sampling Status of a Stream	21
     3.1.3   Elevation at Transect A	24
     3.1.4   Sampling During or After Rain Events	24
     3.1.5   Site Photographs	24
   3.2    LAYING OUT THE SAMPLING REACH	25
     3.2.1   Sliding the Reach	28
   3.3    MODIFYING SAMPLE PROTOCOLS FOR HIGH OR Low FLOWS	29
     3.3.1   Streams with Interrupted Flow	29
     3.3.2   Braided Rivers and Streams	30
                                                                                                     H
4    WATER CHEMISTRY / CHLOROPHYLL-A SAMPLE COLLECTION AND PRESERVATION	32         z
   4.1    IN SITU MEASUREMENTS OF DISSOLVED OXYGEN, pH, TEMPERATURE, AND CONDUCTIVITY	32         z
     4.1.1   Summary of Method	32         8
     4.1.2   Equipment and Supplies	32         Q
     4.1.3   Sampling Procedure	33         i-y
   4.2    WATER CHEMISTRY SAMPLES	35
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               4.2.1   Summary of Method	35
               4.2.2   Equipment and Supplies	35
               4.2.3   Water Chemistry and Chlorophyll-a Sampling Procedure	35

          5    MICROCYSTINS	38

            5.1    SUMMARY OF METHOD	38
            5.2    EQUIPMENT AND SUPPLIES	38
            5.3    SAMPLING PROCEDURE	38
          6    BENTHIC MACROINVERTEBRATES	40

            6.1    SUMMARY OF METHOD	40
            6.2    EQUIPMENT AND SUPPLIES	40
            6.3    SAMPLING PROCEDURE	42
            6.4    SAMPLE PROCESSING IN FIELD	45

          7    PERIPHYTON	48

            7.1    SUMMARY OF METHOD	48
            7.2    EQUIPMENT AND SUPPLIES	48
            7.3    SAMPLING PROCEDURE	48
            7.4    SAMPLE PROCESSING INTHE FIELD	50

          8    PHYSICAL HABITAT CHARACTERIZATION	51

            8.1    EQUIPMENT AND SUPPLIES	51
            8.2    SUMMARY OF METHODS APPROACH	52
            8.3    COMPONENTS OF THE HABITAT CHARACTERIZATION	52
            8.4    WORKFLOW FOR THE PHYSICAL HABITAT COMPONENTS	53
               8.4.1   Thalweg Profile and Large Woody Debris Tally	53
               8.4.2   Channel/Riparian Cross-Sections	53
               8.4.3   Channel Constraint and Torrent Evidence	54
               8.4.4   Stream Discharge	54
            8.5    HABITAT SAMPLING LOCATIONS WITHIN THE REACH	54
               8.5.1   Thalweg Profile and Large Woody Debris Measurements	57
            8.6    CHANNELAND RIPARIAN MEASUREMENTS AT CROSS-SECTION TRANSECTS	64
               8.6.1   Slope and Bearing	64
            8.7    SUBSTRATE SIZE AND CHANNEL DIMENSIONS	70
               8.7.1   Bank Characteristics	 73
            8.8    CANOPY COVER MEASUREMENTS	80
            8.9    VISUAL RIPARIAN ESTIMATES	81
               8.9.1   Riparian Vegetation Structure	81
            8.10   INSTREAM FISH COVER, ALGAE, AND AQUATIC MACROPHYTES	83
            8.11   HUMAN INFLUENCE	84
            8.12   CROSS-SECTION TRANSECTS ON SIDE CHANNELS	86
            8.13   CHANNEL CONSTRAINT, DEBRIS TORRENTS, RECENT FLOODS, AND DISCHARGE	88
               8.13.1    Channel Constraint	88
               8.13.2    Debris Torrents and Recent Major Floods	91
£          8.14   STREAM DISCHARGE	94
•f]             8.14.1    Velocity-Area Procedure	94
^             8.14.2    Timed Filling Procedure	97
O             8.14.3    Neutrally Buoyant Object Procedure	98

O        9    FECAL INDICATOR (ENTEROCOCCI)	100
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m          9.1    SUMMARY OF METHOD	100
i—          9.2    EQUIPMENT AND SUPPLIES	100


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  9.3    SAMPLING PROCEDURE	100
  9.4    SAMPLE PROCESSING INTHE FIELD	101

10   FISH ASSEMBLAGE	102

  10.1   SUMMARY OF METHOD	102
  10.2   EQUIPMENT AND SUPPLIES	105
  10.3   SAMPLING PROCEDURES	105
     10.3.1    Irruptive Species	106
     10.3.2    Small Wadeable Streams	109
     10.3.3    Medium and Large Wadeable Streams	Ill
  10.4   SEINING	115
  10.5   PROCESSING FISH	120
     10.5.1    Identification and Tallying	120
     10.5.2    Unknown Specimens	120
     10.5.3    Photovouchering	124
     10.5.4    Preparing Preserved Voucher Specimen Samples	124
     10.5.5    Preserving Voucher Specimen Samples	125
     10.5.6 Processing Unknown/Range Extension (UNK/RNG) Voucher Samples	128
     10.5.6    Processing QA  Voucher Samples	128

11   FISH TISSUE PLUG SAMPLING METHODS	133

  11.1   METHOD SUMMARY	133
  11.2   EQUIPMENT AND SUPPLIES	133
  11.3   SAMPLE COLLECTION PROCEDURES	134

12   WHOLE FISH SAMPLING METHOD	137

  12.1   METHOD SUMMARY	137
  12.2   EQUIPMENT AND SUPPLIES	137
  12.3   SAMPLING PROCEDURES	138

13   FINAL SITE ACTIVITIES	143

  13.1   OVERVIEW OF FINAL SITE ACTIVITIES	143
  13.2   GENERAL SITE ASSESSMENT	144
     13.2.1    Elevation at Transect K	144
     13.2.2    Watershed Activities and Disturbances Observed	144
     13.2.3    Site Characteristics	144
     13.2.4    General Assessment	144
  13.3   PROCESSING THE FECAL INDICATOR, CHLOROPHYLL-/*, AND PERIPHYTON SAMPLES	147
     13.3.1    Equipment and Supplies (Fecal Indicator Filtering)	147
     13.3.2    Procedures for Processing the Fecal Indicator Sample	147
     13.3.3    Equipment and Supplies (Chlorophyll-afrom Water Sample Filtering)	149
     13.3.4    Procedures for Processing the Chlorophyll-a Water Sample	149
     13.3.5    Equipment and Supplies (Periphyton Sample)	150
     13.3.6    Procedures for Processing the Periphyton Samples	150
  13.4   DATA FORMS AND SAMPLE INSPECTION	154
  13.5   LAUNCH SITE CLEANUP	154         £
                                                                                                       z
14   FIELD QUALITY CONTROL	155         £
                                                                                                       Z
  14.1   REVISIT SAMPLING OVERVIEW	155         O
  14.2   REVISIT SAMPLING SITES	155         u.
  14.3   FIELD EVALUATION AND ASSISTANCE VISITS	156         ^
     14.3.1    Specifications for QC Assurance Field Assistance Visits	156         m
  14.4   REPORTING	157         i—


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           Version 1.0, May 2013                                                                 Wadeable

           15   REFERENCES	159

           APPENDIX A     LIST OF EQUIPMENT AND SUPPLIES	A-l

           APPENDIX B     SAMPLE FORMS	B-l

           APPENDIX C     SHIPPING AND TRACKING GUIDELINES	C-l

           APPENDIX D     COMMON & SCIENTIFIC NAMES OF FISHES OF THE UNITED STATES	D-l

           LIST  OF FIGURES

           Figure 1.1 Example NRSASite Maps	4
           Figure 1.2 Example Sample Labels for Sample Tracking and Identification	7
           Figure 2.1 Field Sampling Scenario for Wadeable Sites	9
           Figure 2.2 Overview of Base Site Activities	10
           Figure 3.1 Verification Form (front)	22
           Figure 3.2 Verification Form (back)	27
           Figure 3.3 Sampling Reach Features for a Wadeable Site	28
           Figure 4.1 Field Measurement Form	34
           Figure 4.2 Sample Collection Form (front)	37
           Figure S.lSample Collection Form (back)	41
           Figure 6.2 Benthic Macroinvertebrate Collection at Wadeable Sites	42
           Figure 6.3 Transect Sample Design for Collecting Benthic Macroinvertebrates at Wadeable Sites	43
           Figure 8.1 Reach  Layout for Physical Habitat Measurements (plan view)	56
           Figure 8.2Thalweg Profile and Woody Debris Form	60
           Figure 8.3 Large Woody Debris Influence Zones (modified from Robison and Beschta, 1990)	64
           Figure 8.4 Slope and Bearing Form	67
           Figure 8.5 Measurements of Bearing Between Transects	69
           Figure 8.6 Channel Slope Measurement using a Clinometer	69
           Figure 8.7 Substrate Sampling Cross-Section	71
           Figure 8.8 Channel Riparian Cross-Section Form	74
           Figure 8.9 Determining Bank Angle Under Different Types of Bank Conditions (A) typical, (B) incised
           channel, (C) undercut bank, and (D) overhanging  bank	76
           Figure 8.10 Schematic Showing Relationship Between Bankfull Channel and Incision	78
           Figure 8.11 Determining Bankfull and Incision  Heights for (A) Deeply Incised Channels, and (B) Streams in
           Deep V Shaped Valleys (Stick figure included for scale)	79
           Figure 8.12 Schematic of Modified Convex Spherical Canopy Densiometer	80
           Figure 8.13 Riparian Zone and Instream  Fish Cover Plots for a Stream Cross-Section Transect	82
           Figure 8.14 Proximity Classes for Human Influences in Wadeable Streams	85
           Figure 8.15 Riparian and Instream Fish Cover Plots for a Stream with Minor and Major Side Channels .... 86
           Figure 8.16 Channel/Riparian Cross-Section Form for an Additional MajorSide Channel Transect	87
           Figure 8.17 Channel Constraint Form	90
           Figure 8.18 Types of Multiple Channel Patterns	91
           Figure 8.19 Torrent Evidence Assessment Form	93
           Figure 8.20 Layout of Channel Cross-Section for Obtaining Discharge Data by the Velocity-Area Procedure
           	94
           Figure 8.21 Discharge Form, showing data recorded for All Discharge Measurement Procedures	96
           Figure 8.22 Use of a Portable Weir in Conjunction with a Calibrated Bucket to Obtain an Estimate of
 t2        Stream  Discharge	97
 ^        Figure 10.1 Fish Gear and Sampling Information Form (front)	103
 52        Figure 10.2 Fish Collection Form	104
 11
 u_        Figure 10.3 Reach Layouts for Fish Sampling at Wadeable Sites	108
 ~;        Figure 10.4 Seining Information Form	119
 ^        Figure 10.5 Unknown/Range Extension Voucher Sample Labels and Voucher Specimen Tags	126
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Figure 10.6 Fish Gear and Sampling Information Form (back)	127
Figure 10.7 QA Voucher Sample Labels and Voucher Specimen Tags	129
Figure 10.8 Fish Identification and Count Update Form	131
Figure 11.1 Whole Fish Tissue Collection Form	142
Figure 13.1 Final Site Activities Summary	143
Figure 13.2 Site Assessment Form (front)	145
Figure 13.3 Site Assessment Form (back)	146
Figure 14.1Summary of the Repeat Sampling Design	155


LIST OF  TABLES

Table 1.1 Summary Table of Indicators for all NRSA 2013/14 Sites	2
Table 1.2 Guidelines for Recording Field  Measurements and Tracking Information	5
Table 2.1 Stock Solutions, Uses, and Methods for Preparation	12
Table 2.2 Post-sampling Equipment Care	13
Table 2.3 General Health and Safety Considerations	15
Table 2.4 General Safety Guidelines for Field Operations	17
Table 3.1 Equipment and Supplies: for Site Verification	20
Table 3.2 Procedure: Site Verification	23
Table 3.3 Guidelines to Determine the Influence of Rain Events	24
Table 3.4 Procedure: Laying Out the Sampling Reach at Wadeable Sites	25
Table 3.5 Procedure: Sliding the Sampling Reach	28
Table 3.6 Reach Layout Modifications for Interrupted Streams	30
Table 3.7 Procedure: Modifications for Braided Rivers and Streams	31
Table 4.1 Equipment and Supplies: DO, pH, Temperature, and Conductivity	32
Table 4.2 Procedure: Temperature, pH, Conductivity and Dissolved Oxygen	33
Table 4.3 Equipment and Supplies: Water Chemistry Sample Collection and Preservation	35
Table 4.4 Procedure: Water Chemistry and Chlorophyll-o Sample Collection (Wadeable Sites)	36
Table 5.1 Equipment and Supplies: Microcystin	38
Table 5.2 Procedure: Microcystin Sample Collection (Wadeable Sites)	39
Table 6.1 Equipment and Supplies: Benthic Macroinvertebrate Collection at Wadeable Sites	40
Table 6.2 Procedure: Benthic Macroinvertebrates (Wadeable Sites)	44
Table 6.3 Procedure: Compositing Samples for Benthic Macroinvertebrates (Wadeable Sites)	46
Table 7.1 Equipment and Supplies: Periphyton (Wadeable Sites)	48
Table 7.2 Procedure: Collecting Composite Index Samples of Periphyton (Wadeable Sites)	49
Table 8.1 Equipment and Supplies: Physical Habitat	51
Table 8.2 Summary of Components of Physical Habitat Characterization at Wadeable Sites	52
Table 8.3 Procedure: Thalweg Profile	58
Table 8.4 Channel Unit Categories	62
Table 8.5 Procedure: Tallying Large Woody Debris	63
Table 8.6 Procedure: Obtaining Slope  Data	65
Table 8.7 Modified  Procedure: Obtaining Slope Data (without Surveyor's Level)	68
Table 8.8 Procedure: Obtaining Bearing Data	70
Table 8.9 Procedure: Substrate Measurement	72
Table 8.10 Procedure: Measuring Bank Characteristics	75
Table 8.11 Procedure: Canopy Cover Measurements	80
Table 8.12 Procedure: Characterizing Riparian Vegetation Structure	82          ^
Table 8.13 Procedure: Estimating Instream Fish Cover	84          ^J
                                                                                                       CO
Table 8.14 Procedure: Estimating Human Influence	85          <
Table 8.15 Procedure: Assessing Channel Constraint	88          Li-
Table 8.16 Procedure: Determining Stream Discharge -Velocity-Area	95          |_
Table 8.17 Procedure: Determining Stream Discharge-Timed Filling	98          ^


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          Table 8.18 Procedure: Determining Stream Discharge - Neutrally Buoyant Object	98
          Table 9.1 Equipment and Supplies:  Fecal Indicator Sampling (Wadeable Sites)	100
          Table 9.2 Procedure: Fecal Indicator (Enterococci) Sample Collection (Wadeable Sites)	100
          Table 10.1 Equipment and Supplies: Fish Collection (Wadeable Sites)	105
          Table 10.2 Summary of Wadeable Fishing Protocols	106
          Table 10.3 Procedure: Electrofishing (Small Wadeable Streams)	109
          Table 10.4 Procedure: Electrofishing (Medium and Large Wadeable Sites)	112
          Table 10.5 Procedure: Seining (Wadeable Sites)	115
          Table 10.6 Procedure: Processing Fish (Wadeable Sites)	121
          Table 10.7 Procedure: Processing Unknown/Range Extension (UNK/RNG) Voucher Samples	130
          Table 10.8 Procedure: Processing QA Voucher Samples	132
          Table 11.1 Equipment and Supplies: Fish Tissue Plug Sample	133
          Table 11.2 Recommended Target and Alternate Species for Fish  Tissue Plug Collection	134
          Table 11.3 Procedure: Fish Tissue Plug Samples	135
          Table 11.4 Equipment and Supplies: Whole Fish Tissue Sample Collection	138
          Table 11.5 Recommended Target Species for Whole Fish Tissue Collection	139
          Table 11.6Sampling Procedures for Whole Fish Tissue Samples	140
          Table 13.1 Equipment and Supplies: Fecal Indicator Sample Processing	147
          Table 13.2 Procedure: Fecal Indicator Sample Processing	147
          Table 13.3 Equipment and Supplies:r Chlorophyll-o Processing	149
          Table 13.4 Procedure: Chlorophyll-o Sample Processing	149
          Table 13.5 Equipment and Supplies: Periphyton Samples	150
          Table 13.6 Procedure: ID/Enumeration Samples of Periphyton	151
          Table 13.7 Procedure: Preparing Chlorophyll Samples of Periphyton	152
          Table 13.8 Procedure: Preparing Ash-Free Dry Mass (AFDM) Samples of Periphyton	153
          Table 14.1 General Information Noted During Field Evaluation	157
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National Rivers and Streams Assessment 2013/14
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                                                      Field Operations Manual
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ACRONYMS/ABBREVIATIONS
AFDM
ANC
Ca
CAS
Cl
CPR
CSDGM
CWA
DELT
DO
DOC
Dl
DQO
EMAP
EPA
FGDC
FOIA
FOM
CIS
GPS
HQ
IBI
IQG
IM
ITIS
K
LIMS
LOM
LRL
LWD
ISO
MDL
Mg
MMI
MQO
MSDS
Na
NABS
NAD
NARS
NAWQA
NCCA
NELAP
NERL
NHD
NH3
NH4
NIST
NLA
NLCD
NO2
Ash-Free Dry mass
Alkalinity
Calcium
Chemical Abstract Service
Chloride
Cardiopulmonary Resuscitation
Content Standards for Digital Geospatial Metadata
Clean Water Act
Deformities, Eroded Fins, Lesions and Tumors
Dissolved Oxygen
Dissolved Organic Compound
Di-ionized Tap Water
Data Quality Objective
Environmental Monitoring and Assessment Program
Environmental Protection Agency
Federal Geographic Data Committee
Freedom of Information Act
Field Operations Manual
Geographic Information System
Global Positioning Device
Head Quarters
Index of Biotic Integrity
Information Quality Guideline
Information Management
Integrated  Taxonomic Information System
Potassium
Laboratory Information Management System
Lab Operations  Manual
Laboratory Recording Levels
Large Woody Debris
International Organization for Standardization
Method Detection Levels (limit)
Magnesium
Multimetric Indicators
Measurement Quality Objective
Methods Safety Data Sheets
Sodium
North American Benthic Society
North American Datum
National Aquatic Resources Survey
National Water-Quality Assessment Program
National Costal Condition Assessment
National Environmental Laboratory Accreditation Program
New England Regional Laboratory
National Hydrology Database
Ammonia
Ammonium
National Institute of Standards
National Lakes Assessment
National Land Cover Dataset
Nitrite
LJJ
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CO
CO
CO
^
>
z
O
C£
U
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          Version 1.0, May 2013
                                                        Field Operations Manual
                                                                    Wadeable
 LJJ
 C£
 CD
 CD
 CO
 ^
 >
 z
 O
 U
          NO3
          NRC
          NRSA
          NWCA
          0/E
          OSHA
          OW
          PC
          PD
          PDE
          PDF
          PP
          PPT
          PRISM
          PTD
          QA
          QAPP
          QA/QC
          QCS
          QRG
          RL
          RTE
          RVT
          SAS
          SDTD
          SEG
          SiO2
          S04
          SOP
          SQL
          Std
          STORE!
          TL
          TOC
          TN
          TP
          TSS
          UNK/RNG
          USGAO
          USGS
          WED
          WSA
          WQX
Nitrate
National Research Council
National Rivers and Streams Assessment
National Wetland Condition Assessment
"Observed" over "Expected"
Occupational Safety and Health Administration
Office of Water
Personal Computer
Percent Difference
Percent Difference in Enumeration
Personal Flotation Device
Polypropylene
Parts per thousand
Parameter-Elevation Regressions on Independent Slopes Model
Percent Taxonomic Disagreement
Quality Assurance
Quality Assurance Protection Plan
Quality Assurance/Quality Control
Quality Check Solution
Quick Reference Guide
Reporting Limit
Rare, Threatened and Endangered
Revisit
Statistical Analysis System
Spatial Data Transfer Standard
Site Evaluation Guideline
Silica
Sulfate
Standard Operating Procedures
Standard Query Language
Standard
Storage and Retrieval Data Warehouse
Total Length
Total Organic Carbon
Total Nitrogen
Total Phosphorus
Total Suspended Solids
Unknown Range Extension
United States General Accounting Office
United States Geological Survey
Western Ecology Division
Wadeable Streams Assessment
Water Quality Exchange
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National Rivers and Streams Assessment 2013/14
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Field Operations Manual
            Wadeable
DISTRIBUTION LIST
This Field Operation Manual (FOM) and associated manuals or guidelines will be distributed to
the following EPA, senior staff participating in the NRSA and to State Water Quality Agencies or
cooperators who will perform the field sampling operations. The Quality Assurance (QA) Officers
will distribute the  Quality  Assurance Project  Plan (QAPP)  and  associated documents  to
participating project staff at their respective facilities and to the project contacts at  participating
laboratories, as they are determined.

Ellen Tarquinio
NRSA Project Leader
Sarah Lehman
NRSA Project QA Officer
Virginia Fox-Norse
OWOW Quality Assurance Officer
Steven G. Paulsen
EPA ORD Technical Advisor
Marlys Cappaert
SRA International Inc.
NARS Information Management
Coordinator
Chris Turner, GLEC, Inc.
Contract Logistics Coordinator.
Leanne Stahl
OST Fish Tissue Coordinator
Robert Shippen
OST QA Fish Tissue Officer


Tom Faber, Region 1
Darvene Adams, Region 2
Louis Reynolds, Region 3
David Melgaard, Region 4
Mari Nord, Region 5
Mike Schaub, Region 6
Gary Welker, Region 7
Karl Hermann, Region 8
Terry Fleming, Region 9

tarquinio.ellen@ epa.gov
202-566-2267
lehmann.sarah@epa.gov
202-566-1379
fox-norse.virginia@epa.gov
202-566-1266
paulsen.steve@epa.gov
541-754-4428
cappaert.marlvs@epa.gov
541-754-4467
541-754-4799 (fax)
cturner@glec.com
715-829-3737
Stahl.leanne@epa.gov
202-566-0404
Shippen. Robert@epa.gov
202-566-0391
Regional Monitoring Coordinator
faber.tom@epa.gov
617-918-8672
adams.darvene@epa.gov
732-321-6700
revnolds.louis@epa.gov
304-234-0244
melgaard.david@epa.gov
404-562-9265
nord.mari@epa.gov
312-353-3017
schaub.mike@epa.gov
214-665-7314
welker.garv@epa.gov
913-551-7177
hermann.karl@epa.gov
303-312-6228
fleming.terrence@epa.gov
415-972-3462

U.S. EPA Office of Water
Office of Wetlands, Oceans, and Watersheds
Assessment and Watershed Protection Division
Washington, DC
U.S. EPA Office of Water
Office of Wetlands, Oceans, and Watersheds
Assessment and Watershed Protection Division
Washington, DC
U.S. EPA Office of Water
Office of Wetlands, Oceans, and Watersheds
Oceans and Coastal Protection Division
Washington, DC
Freshwater Ecology Branch Western Ecology
Division, NHEERL, ORD, EPA
200 S.W. 35th St. Corvallis, OR 97330
Computer Science Corporation
200 S.W. 35th Street
Corvallis, OR 9733
Great Lakes Environmental Center
739 Hastings Street
Traverse City, Ml 49686
U.S. EPA Office of Water
Office of Science and Technology
Washington, DC
U.S. EPA Office of Water
Office of Science and Technology
Washington, DC
s
U.S. EPA- Region 1
11 Technology Drive North Chelmsford, MA
01863-2431
USEPA- Region II
2890 Woodbridge Ave Edison, NJ 08837-3679
U.S. EPA -Region III
303 Methodist Building Wheeling WV 26003
U.S. EPA -Region IV
61 Forsyth Street, S.W. Atlanta, GA 30303-8960
U.S. EPA - Region V
77 West Jackson Blvd Chicago, IL 60604-3507
U.S. EPA -Region VI
1445 Ross Ave -Ste 1200 Dallas, TX 75202-2733
U.S. EPA -Region VII
901 North Fifth Street Kansas City, KS 66101
U.S. EPA- Region VIM
1595 Wynkoop St .Denver, CO 80202-1129
U.S. EPA -Region IX
75 Hawthorne Street San Francisco, CA 94105
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                                          Field Operations Manual
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               Gretchen Hayslip, Region 10
havslip.gretchen(5)epa.gov
     206-553-1685
       U.S. EPA-Region X,
1200 Sixth Avenue Seattle, WA 98101
 CD
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1      BACKGROUND
This manual describes field protocols and daily operations for crews to use in the National Rivers
and Streams Assessment (NRSA) 2013/14. The NRSA is a probability-based survey of our
Nation's rivers and streams and is designed to:

   •  Assess the condition of the Nation's rivers and streams;

   •  Evaluate changes in condition from the 2008/09 NRSA; and

   •  Help build State and Tribal capacity for monitoring and assessment and promote
       collaboration across jurisdictional  boundaries.
This is one of a series of water assessments being conducted by states, tribes, the U.S.
Environmental Protection Agency (EPA), and other partners. In addition to rivers and streams,
the water assessments will also focus on coastal waters, lakes, and wetlands in a revolving
sequence. The purpose of these assessments is to generate statistically valid reports on the
condition of our Nation's water resources  and identify key stressors to these systems.

1.1    Survey Design
EPA selected sampling  locations using a probability based  survey design. The design is an
unequal probability design that selects 900 sites classified  as Strahler order 1-4 and 900 sites
classified as Strahler order 5th and above. To evaluate change from the 2008/09 NRSA, 420 of
the 900 l-4th order sites are resampled from the 2008/09 NRSA and 390, 5th orders and above
sites are revisits from the 2008-2009 NRSA. Approximately 10%, or 200, of the total NRSA sites
are scheduled for repeated sampling (revisit sites) in the same field season. The sample frame
was derived from the National Hydrography Dataset (NHD), NHD-Plus, from 1:100,000 scale
maps. Additional details on the NRSA survey design are found in the National Rivers and Streams
Assessment Survey Design: 2013-2014 documents.

1.2    Target Population and Index Period
The target population consists of all streams and rivers within the 48 contiguous states that
have flowing water during the study index period. This includes major rivers, and small streams.
Sites must have > 50%  of the reach length with standing water. Sites with water in less than 50%
of the reach length must be dropped. All sites must be sampled during base flow conditions.
The target population excludes:

   •  Tidal rivers and streams up to head of salt (defined as < .OSppt for this study).

   •  Run-of-the-river ponds and reservoirs with greater than 7 day residence time.

   •  The study index period extends from:
           o  Beginning of June through end of September for most regions
           o  Sites in the select ecoregions or States can be sampled starting in the end of
              April with approval from the EPA Project Coordinator
                                                                                              Q
Please refer to the Site Evaluation Guidelines (EPA-841-B-012-008) and the NRSA Web site               §
(http://www.epa.gov/owow/riverssurvey/index.html) for  more detailed information on the              §
target population and exclusion criteria.                                                           ^
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         1.3    Replacing Sites
         All base sites must be evaluated for sampling. If a stream or river site is determined to be
         unsampleable, it must be replaced by another site within the state in the same category. The six
         general categories for each state are:

             •   NRSA 2008-2009 resample 1 - 4th Strahler order sites.

             •   NRSA 2008-2009 resample 5th and above Strahler order sites.

             •   Small Stream- new 1-3 Strahler order sites.

             •   Large Streams- new 3-5 Strahler order sites.

             •   Major Rivers- new 5 and above order sites. Rivers identified as major rivers or
                additional rivers in the book: Rivers of North America.

             •   Other Rivers- new 5 and above order sites that are not considered Major Rivers.
         Please refer to the Site Evaluation Guidelines (EPA-841-B-12-008) for more detailed information.

         1.4    Selection of NRSA Indicators
         As part of the indicator selection process, EPA worked with state and tribal partners and other
         partners through technical conferences and  indicator teleconferences. The EPA formed a
         National Rivers and Streams Assessment Steering Committee with state, tribal and regional
         representatives to provide feedback and evaluate core and supplemental indicators to be
         included in the 2013/14 field season. Key evaluation criteria included indicator applicability on a
         national scale, the ability of an indicator to reflect various aspects of ecological condition,
         repeatability, and cost-effectiveness. The core indicators build upon the work done in the NRSA
         2008/09. They have been sampled and analyzed on the national scale and have a known
         applicability to Clean Water Act (CWA) programs. Supplemental indicators were selected based
         on feedback from the Steering Committee and decisions by EPA management. Supplemental
         indicators are either in the research phase and their applicability is still being assessed for CWA
         programs or this is the first time they will be sampled at a national scale. For field sampling
         purposes, there is no distinction between core and supplemental indicators. Indicators that are
         included in the NRSA 2013/14 are briefly described in Table 1.1.
         Table 1.1 Summary Table of Indicators for all NRSA 2013/14 Sites
                   Indicator
                              Core or Supplemental
                                    Indicator
          In Situ measurements (pH, DO,    Core Indicator
          temperature, conductivity)
                           Specs/Location in Sampling Reach
                                                     Measurements taken at X site at
                                                     midchannel; readings are taken at 0.5 m
                                                     depth, or mid-depth if water depth is less
                                                     than 1 meter.
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          Water chemistry (TP, TN [NH4,    Core Indicator
          NO3), basic anions and cations,
          alkalinity [ANC], DOC, TOC, TSS,
          conductivity
                                                     Measurements taken at X site at
                                                     midchannel; Collected from a depth of
                                                     0.5 m, or mid-depth if water depth is less
                                                     than 1 meter.
Chlorophyll-a
Core Indicator
Collected as part of water chemistry and
periphyton samples

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 Microcystin

 Periphyton



 Benthic macroinvertebrate
 assemblage (Littoral)


 Fish Assemblage


 Physical habitat assessment


 Fecal indicator (Enterococci)


 Fish Tissue Plug



 Whole Fish Tissue
Supplemental Indicator

Core Indicator



Core Indicator



Core Indicator


Core Indicator


Supplemental Indicator


Supplemental Indicator
Supplemental Indicator at
select sites
Collected from index site

Collected from 11 locations
systematically placed at each site and
combined into a single composite sample

Collected from 11 locations
systematically placed at each site and
combined into a single composite sample

Sampled throughout the sampling reach
at specified locations

Measurements collected throughout the
sampling reach at specified locations

Collected at the last transect one meter
off the bank at .3 m depth

Target species collected throughout the
sampling reach as part offish
assemblage sampling

Target species collected throughout the
sampling reach as part offish
assemblage sampling
1.5     Supplemental Material to the Field Operations Manual
The Field Operations Manual describes field protocols and daily operations for crews to use in
the NRSA. Following these detailed protocols will ensure consistency across regions and
reproducibility for future assessments. Before beginning sampling at a site, crews should
prepare a packet for each site containing pertinent information to successfully conduct
sampling. This includes a road map and set of directions to the site, topographic maps,
landowner access forms, sampling permits (if needed), site evaluation forms and other
information necessary to ensure an efficient and safe sampling day.
Site maps have been  provided to assist in the site evaluation process. Three maps are available:
an aerial image, topographic map, and road map, see Figure 1.1. These maps provide an overlay
of the NHD waterbody layer with the coordinate and label for the x-site and/or waterbody, if
available. Each site is symbolized by the panel the site is considered  within (see Section 1.3).
Other important information that may assist in site evaluation is included on the map including:
state, EPA region, latitude, longitude, ownership, and stream order.  These maps will be helpful
in the planning and preparation for visiting and sampling a particular NRSA 2013/14 site. These
maps will become part of your site packet.
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            2013-2014 National Rivers and Streams Assessment
                  Oklahoma Site: OKRM-1006
                                        2013-2014 National Rivers and Streams Assessment
                                               Oklahoma Site: OKRM-1006
                                                                    2013-2014 National Rivers and Streams Assessment
                                                                          Oklahoma Site: OKRM-1006
Latitude (DD):
Longitude (DD):
Stream Order:
Ownership:
Reacticode:

Scale:
Imagery Date:
Imagery Source:
                                          35.314465
                                         -94.466461
                                               5
                                         NonFederal
                                     11110105000002

                                          1:55,000
                                             2010
                                    National GeoTOPO!
         Figure 1.1 Example NRSA Site Maps
         Field crews will also receive a Quick Reference Guide (QRG) that contains tables and figures
         summarizing field activities and protocols from the Field Operations Manual (FOM). This
         waterproof handbook will be the primary field reference used by field crews after reading the
         FOM and completing the required field training session. The field crews are also required to
         keep the field operations manual available in the field for reference and for possible protocol
         clarification.
         Quality assurance is a required element of all  EPA-sponsored studies that involve the collection
         of environmental data (USEPA 2000a, 2000b). Field crews will be provided a copy of the
         integrated Quality Assurance and Project Plan (QAPP). The QAPP contains more detailed
         information regarding quality assurance/quality control (QA/QC) activities and procedures
         associated with general field operations, sample collection, measurement data collection for
         specific indicators, and data reporting activities. For more information on the Quality Assurance
         procedures, refer to the National Rivers and Streams Assessment: Quality Assurance Project Plan
         (EPA 841-B-012-007).
         Related NRSA documents include the following: National Rivers and Streams Assessment:
         Quality Assurance Project Plan  (EPA 841-B-12-007), National Rivers and Streams Assessment:
         Site Evaluation Guidelines (EPA841-B-012-008), and National Rivers and Streams Assessment:
         Laboratory Methods Manual (EPA 841-B-012-010). These documents are  available at:
         http://www.epa.gov/owow/riverssurvey/index.html.
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1.6     Recording Data and Other Information

All samples need to be identified and tracked, and associated information for each sample must
be recorded. To assist with sample identification and tracking, labels are preprinted with sample
ID numbers (Figure 1.2).

It is imperative that field and sample information be recorded accurately, consistently, and
legibly. The cost of a  sampling visit coupled with the short index period severely limits the ability
to resample a site if the initial information recorded was inaccurate or illegible. Guidelines for
recording field measurements are presented in Table 1.2.
Table 1.2 Guidelines for Recording Field Measurements and Tracking Information
                                      Field Measurements
Data Recording
    •   Record measurement values and observations on data forms preprinted on
        water-resistant paper.
    •   Use No. 2 pencil only (fine-point indelible markers can be used if necessary)
        to record information on forms.
    •   Record data and information using correct format as provided on data forms.
    •   Be sure to accurately record site IDs and sample numbers. For all primary
        sampling visits indicate the event as Visit 1. For revisit sites use Visit 2 to
        indicate the second sampling event during the same season.
    •   Print legibly (and as large as possible). Clearly distinguish letters from
        numbers (e.g., 0 versus O, 2 versus Z, 7 versus T or F, etc.), but do not use
        slashes.
    •   In cases where information is recorded repeatedly on a series of lines (e.g.,
        physical  habitat characteristics), do not use "ditto marks" (") or a straight
        vertical line. Record the information that is repeated on the first and last
        lines, and then connect these using a wavy vertical line.
    •   When recording comments, print or write legibly. Make notations in
        comments field only; avoid marginal notes. Be concise, but avoid using
        abbreviations or "shorthand" notations. If you run out of space, attach a
        sheet of  paper with the additional information, rather than trying to squeeze
        everything into the space provided on the form.
Data Qualifiers
(Flags)
Use only defined flag codes and record on data form in appropriate field.
      K   =    Measurement not attempted or not recorded.
      Q  =    Failed quality control check; remeasurement not possible.
      U   =    Suspect measurement; remeasurement not possible.
      fn  =    Miscellaneous flags (n = 1, 2, etc.) assigned by a field crew during a
                 particular sampling visit (also used for qualifying samples).
Explain reason for using each flag in comments section on data form.
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          Sample Labels
       Use adhesive labels with preprinted ID numbers and follow the standard
       recording format for each type of sample.
       Use a pencil to record information on label. Cover the completed label with
       clear tape.
       Record sample ID number from label and associated collection information
       on sample collection form preprinted on water-resistant paper.
       Sample IDs from a single label sheet are in  sequential order. Do not mix
       labels from different sheets.
          Sample Qualifiers
          (Flags)
             Sample Collection and Tracking
Use only defined flag codes and record on sample collection form in appropriate
     field.
     K   =    Sample not collected or lost before shipment; resampling not
               possible.
     U   =    Suspect sample (e.g., possible contamination, does not meet
               minimum acceptability requirements, or collected by non-standard
               procedure).
     Fn  =    Miscellaneous flags (n=l, 2, etc.) assigned by a field crew during a
               particular sampling visit (also used for field measurements).
Explain reason for using flags in "Comments" on sample collection form.
          Review of Labels
          and Data
          Collection Forms
       Compare information recorded on labels and sample collection form for
       accuracy before leaving site.
       Review labels and data collection forms for accuracy, completeness, and
       legibility before leaving site.
       The Field Crew Leader must review all labels and data collection forms for
       consistency, correctness, and legibility before transfer to the Information
       Management Center.
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               CHEMISTRY (CHEM)
             Site ID:	
       Dale
              /201   Visit* O1 O2
                    990000
                                           WATER COLUMN CHLOROPHYLL (CHLA)
                                                   Site ID:	
                                             Date:
    I
/201   Visit*: O1 O2
                                                   Volume Filtered: _
                                                           990001
                                                                            ml
PERIPHYTON ASSEMBLAGE ID (PERI)
      Site ID:	
Date:     /    /201    Visit *O1 O2
     Composite Volume: 	mL
             990002
                                                    PERIPHYTON CHLOROPHYLL (PCHL)
                                                          Site ID:
                                                    Date:
                                                           /201
	 Visit*: 01 02
 Volume Filtered:  	ml
        990003
          PERIPHYTON BIOMASS (PBIO)
             Srte ID:	
       Date
         /	/201   Visit* O1 O2
             Volume Fiftered:
                        mL
                          _
                    990004
Dale:     /     /201   Visit* O1 O2
           Jar 1 of
                    990006
            FISH TISSUE PLUG (FPLG)
             Site ID: _ X
                                    0
                                      0
                                      °
Date:
                /     /201   \A«at*:Ol O2
                    99000E
                                                    ALGAL i OXIN (MICX)
                                                   Sit-10:	
                                             Date
                                                             I
	_ Visit* 01 02
Volume Collected: 	mL
        990005
     BENTHIC MACROINVERTEB RATES (BERW)
             Site ID:	
                                             A
                                           3EUTHIC MACROINVERTEBRATES (BETB)
                                                   Srte ID: _
                                             Date:     /    /201    Visit*: O1 O2
                                                    Date:
                                                                  /201
                                                               Jar 1 of
                                                           990007
                                                    OA VOUCHER (VERT)
                                                   Site ID:
                                                      I
                                                           /201   Visit*: 01 02
                                                                 990009
    UNK/RNG VOUCHER (VERT)
      Site ID:	
Date:	/	/201    Visit *O1 O2
                                                      Sample Type:
                                                          Srte ID:
                                                             I
                                                         Sample ID:
                                                           /201   Veil*: 01 O2
         Sampl« Typ«:
             Site ID: _
       Date:     /
              /201   Visit* 01 02
            Sample ID:
                                               Samplt Typ«:
                                                   SHeC:_
                                             Date:     /
         /201   Visit*: 01 02
                                                  Sample ID:
Figure 1.2 Example Sample Labels for Sample Tracking and Identification.
                                                                                                     Q

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         2      INTRODUCTION TO WADEABLE SAMPLING

         2.1    Daily Operations
         Field methods for the NRSA are designed to be completed in one field day for most sites.
         Depending on the time needed for both the sampling and travel for the day, an additional day
         may be needed to complete sampling or for pre-departure and post-sampling activities (e.g.,
         cleaning equipment, repairing gear, shipping samples, and traveling to the next site). Remote
         sites with lengthy or difficult approaches may require more time, and field crews will need to
         plan accordingly.
         A field crew for wadeable sites will typically consist of four people. Any additional crew
         members may either help collect samples, or may remain on the bank to provide logistical
         support. Typically, in wadeable sites, two crew members will work on the "habitat" crew, and
         two or three will work on the "fish" crew.
         A daily field sampling scenario showing how the work load may be split between crew members
         is presented in Figure 2.1. The following sections further define the sampling sequence and the
         protocols for sampling activities.
         Field crews should define roles and responsibilities for each crew member to organize field
         activities efficiently. While crews may choose to allocate resources as they see fit, the sequence
         of sampling events presented in the Figure 2.1 cannot be changed  and is based on the need to
         protect some types of samples from potential contamination and to minimize holding times
         once samples are collected.

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             Wadeable
                                          Whole Crew
Group A Activities:


[
Locate Xsite
Verify site as target
Set up staging area
1

Prepare forms, equipment & supplies

I

Lay out sampling reach (from X-site to Transect A) La
i


BEGIN SAMPLING ACTIVITIES AT TRANSECT A
i

Conduct physical habitat
Characterizations
Including slope & bearing
;
,
Collect benthic samples
:
,
Collect periphyton samples

,
Collect fecal indicator
sample at last transect

k
Measure stream discharge

\



RETURN TO STAGING AREA
1
Preserve benthic samples
prepare for transport

Inspect and clean boat, mo-
tor, & trailerto prevent trans-
fer of nuisance species and
contaminants


*
Filter fecal indicator sample;
prepare for transport

1
Filter chlorophy ll-a sample;
prepare for transport

Prepare periphyton samples
fortransport

I

Clean and organize equipment for loading





1
SHIP SAMPLES



Calibrate multi-parameter probe
I
Activities :


/ out sampling reach (from X-site to Transect K)
1
RETURN TO TRANSECT F (X-SITE)
1
Measure in situ temperature
pH, DO, & conducts iy
I
Collect water chemistry,
chlorophy ll-a and microcystin
samples
1
TRAVEL TO TRANSECT A
1
Conduct fish assessment
1
Collect fish tissue samples
1
RETURN TO STAGING AREA
1
Prepare fish issue samplesf or
transport
5







Review data forms for completeness
I
Inventory supplies for next sampling event
Request additional supplies if needed
1

Report sampling event through Site and Sam-
ple Status Form




Figure 2.1 Field Sampling Scenario for Wadeable Sites
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         2.2    Base Site Activities

         Field crews conduct a number of activities at their base site (i.e., office or laboratory, camping
         site, or motel). These include tasks that must be completed both before departure to the site
         and after return from the field (Figure 2.2). Close  attention to these activities is required to
         ensure that the field crews know (1) where they are going, (2) that access is permissible and
         possible, (3) that equipment and supplies are available and in good working order to complete
         the sampling effort, and (4) that samples are packed and shipped appropriately.
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                       PREDEPARTURE ACTIVITIES
             Crew Leader
             •   Prepare daily itinerary
     Crew Members
     •   Instrument checks & calibration
     •   Equipments supplies preparation
Whole Crew
 Site verification
                                 SAMPLE SITE
                       POST-SAMPLING ACTIVITIES
             Crew Leader
             •   Review forms & labels
             •   File status report by email to
                 IM Team
     Crew Members
     •   Filter, preserve & inspect samples
     •   Clean boats/gear with 1-10%
         bleach solution
     •   Make any repairs necessary
     •   Charge or replace batteries
     •   Refuel boats, vehicles, etc.
     •   Obtain ice, dry ice and other con-
         sumables.
     •   Package andshipsamples and
         data forms
         Figure 2.2 Overview of Base Site Activities

         2.2.1   Pre-departure Activities

         Pre-departure activities include the development of a daily itineraries, instrument checks and
         calibration, and equipment and supply preparation. Procedures for these activities are described
         in the following sections.

         2.2.1.1  Daily Itineraries

         The Field Crew Leaders are responsible for developing daily itineraries. This entails compiling
         maps, contact information, copies of permission letters, and access instructions (a "site
10

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packet"). Additional activities include confirming the best access routes, calling the landowners
or local contacts, confirming lodging plans, and coordinating rendezvous locations with
individuals who must meet with field crews prior to accessing a site. Changes in the itinerary
during the week, such as canceling a sampling day, must be relayed by the crew leader to the
Field Logistics Coordinator as soon as possible.

2.2.1.2 Instrument Checks and Calibration
Each field crew must test and calibrate instruments prior to sampling. Calibration can be
conducted prior to departure for the site or at the site, with the exception of dissolved oxygen
(DO) calibration. Because of the potential influence of altitude, DO calibration is to be
performed only at the site. Field instruments include a global positioning system (GPS) receiver,
a multi-probe unit for measuring DO, pH, temperature, and conductivity, and electrofishing
equipment. Field crews should have access to backup instruments if any instruments fail the
manufacturer performance tests or calibrations. Prior to departure, field crews must:

    •  Turn on the GPS receiver and check the batteries. Replace batteries immediately if a
       battery warning is displayed.
    •  Test and calibrate the multi-probe meter. Each field crew should have a copy of the
       manufacturer's calibration and maintenance procedures. All meters should be
       calibrated according to manufacturer specifications provided along with the meter.
       Once a week, crews should check their multi-probe against a Quality Check Solution
       (QCS).
    •  Turn on the electrofishing unit and check the batteries. Be sure to have fully charged
       backup batteries. If using a gas powered electrofishing unit, check the oil and gas supply.

2.2.1.3 Equipment and Supply Preparation
Field crews must check the inventory of supplies and equipment prior to departure using the
equipment and supplies checklists  provided in Appendix A; use of the lists is mandatory. Specific
equipment will be used for wadeable or non-wadeable sites; be sure  to bring both sets of
equipment if you are  unsure what type of site you will be visiting that day. Pack meters, probes,
and sampling gear in such a way as to minimize physical shock and vibration during transport.
Pack stock solutions as described in Table 2.1. Follow the regulations of the Occupational Safety
and Health Administration (OSHA).
Site kits of consumable supplies for each sampling site will be delivered based on the supply             ^
requests each crew submits prior to and during the sampling season. Crews will submit an               ?
electronic request form to order site kits, forms, labels, etc. Site kit requests must be submitted          ^
at least three weeks before sampling is to take place. Field crew leaders MUST provide a                <
tentative schedule in order to receive the site kits. Crews should include in this schedule the           ^
primary fish taxonomist at each site. If your schedule changes, report the change as soon as             <
possible to the Field Logistics Coordinator (Chris Turner, cturner@glec.com, (715)829-3737). The         a
                                                                                                ^f
site kit will include  data forms, labels, sample jars, bottles, and other supplies (see complete list          ^
in Appendix A). The crews must inventory these site kits before departure. Container labels              O
should not be covered with clear tape until all information is completed during sampling at the           -z.
river/stream. Store at least one extra site kit in the vehicle in the event replacement items are           p
needed immediately.                                                                             ^
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         Table 2.1 Stock Solutions, Uses, and Methods for Preparation
           Solution
           Bleach (1%)
                           Clean nets, other gear, and boat.
                                                             Preparation
Add 40 mL bleach to 4 L distilled water.
           QCS Solution*
                           Weekly check of meter calibration
                           A 1:100 dilution of the standard
                           solution (RIGHT) produces a solution
                           with the following theoretical values:
                            pH           6.98
                            Conductivity 75.3 u.S/cm @ 25oC
STANDARD SOLUTION:
KH2PO4
Na2HPO4
Deionized water
Mix to dissolve
                                                                                           3.4022 g
                                                                                            3.5490 g
                                                                                            1000 ml
           10% Buffered     Preservation of periphyton ID sample
           Formalint        and fixing Fish Vouchers
                                                             Formaldehyde (37-40%)
                                                             Distilled water
                                                             NaH2PO4
                                                             Na2HPO4 (anhydrous)
                                                             Mix to dissolve
                                100ml
                                900ml
                                 4.0 g
                                 6.5 g
           95% Ethanol      Preservative for benthic invertebrate
                           samples and fish vouchers.
                                                             No preparation needed (use stock solution as
                                                             is).
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             * QCS or "confidence" solutions can also be purchased pre-mixed from various sources.
             f 10% Buffered Formalin can also be purchased pre-mixed from various sources

          2.2.2   Post Sampling Activities
          Upon return to the launching location after sampling, the crew must review all completed data
          forms and labels for accuracy, completeness, and legibility and make a final inspection of
          samples. If information is missing from the forms or labels, the Field Crew Leader is to provide
          the missing information. The Field Crew Leader is to initial all data forms after review. If
          obtainable samples are missing, the site should be rescheduled for complete sampling. Other
          post sampling activities include: inspection and cleaning of sampling equipment, supply
          inventory/reorder, sample and data form shipment, and communications.

          2.2.2.1  Review Data Forms and Labels
          The field crew leader is ultimately responsible for reviewing all data forms and labels for
          accuracy, completeness, and legibility. Ensure that written comments  use no "shorthand" or
          abbreviations. The data forms must be thoroughly reviewed. Upon completing the review, the
          field crew leader  must initial the field forms to indicate that they are ready to be sent to the
          Information Management Center (a similar review process is used for  electronic forms). Each
          sample label must also be checked for accuracy, completeness, and legibility. The field crew
          leader must cross-check the sample numbers on the labels with those recorded on the data
          forms.

          2.2.2.2  Inspect and Prepare Samples
          All samples need  to be inspected and appropriately preserved and packaged for transport.
          Check that all samples are labeled, and all labels are completely filled in. Check that each label is
          covered with clear plastic tape. Check the integrity of each sample container, and be sure there
          are no leaks. Make sure that all sample containers are properly sealed. Make sure that all
          sample containers are properly preserved for storage or immediate shipment.
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2.2.2.3 Equipment Cleanup and Check
All equipment and gear must be cleaned and disinfected between sites to reduce the risk of
transferring nuisance species and pathogens. Species of primary concern in the U.S. include
Eurasian watermilfoil (Myriophyllum spicatum), zebra mussels (Dreissena polymorpha), New
Zealand mud  snails (Potamopyrgus antipodarum), Myxobolus cerebralis (a sporozoan parasite
that causes salmonid whirling disease), and Batrachochytrium dendrobatidis (a chytrid fungus
that threatens amphibian populations). Field crews must be aware of regional species of
concern, and  take appropriate precautions to avoid transfer of these species. There are several
online resources regarding invasive species,  including information on  cleaning and disinfecting
gear, such as  the Whirling Disease Foundation (www.whirling-disease.org). the USDA Forest
Service (Preventing Accidental Introductions of Freshwater Invasive Species, available from
http://www.fs.fed.us/invasivespecies/documents/Aquatic is  prevention.pdf). and the
California Dept. of Fish and Game (Hosea and Finlayson 2005). General information  about
freshwater invasive species is available from the U.S. Geological Survey Nonindigenous Aquatic
Species website (http://nas.er.usgs.gov), the Protect Your Waters website that is co-sponsored
by the U.S. Fish and Wildlife Service (http://www.protectyourwaters.net/hitchhikers), and the
Sea Grant Program (http://www.sgnis.org).
Handle and dispose of disinfectant solutions properly, and take care to avoid damage to lawns
or other property. Table 2.2 describes equipment care.  Inspect all equipment, including nets,
boat, and trailer, and clean off any plant and animal material. Prior to leaving a site, drain all
bilge water and live wells in the boat. Inspect, clean, and handpick plant and animal remains
from vehicle,  boat, motor, and trailer. Before moving to the next  site, if a commercial car wash
facility is available, wash vehicle, boat, and trailer and rinse thoroughly (hot water pressurized
rinse no soap). Rinse equipment and boat with 1% -10% bleach solution to prevent the spread
of exotics. Note that many organizations now recommend against using felt-soled wading boots
in affected areas due to the difficulty in removing myxospores and mudsnails.

2.2.2.4 Supply Inventory
Once a field day is completed, crews should  inventory and restock supplies as needed. Ensure
that there is a sufficient quantity of site kits to allow sampling at upcoming sites (for at least the
next 2 weeks). Take note of any supplies that are nearing depletion. Also note any items that
may have been lost or damaged during the sampling event. Request additional site kits and/or
supplies as needed via the electronic request form. Requests must be made three weeks before
needed. Note that not all supplies can be replenished by EPA through the Logistics Contractor,
so crews will  need to supply some items themselves.
Table 2.2 Post-sampling Equipment Care
                               Equipment Care after Sampling
1.  Clean for biological contaminants.
Prior to departing site, drain all water from live wells and buckets used to hold and process fish, and drain
all bilge water from the boat.
Inspect motor, boat, trailer, sampling gear, waders, boots, etc. for evidence of mud, snails, plant
fragments, algae, animal remains, or debris, and remove using brushes or other tools.
At the base location, inspect and rinse seines, dip nets, kick nets, waders, and boots with water and dry.
Use one of the procedures below to disinfect gear if necessary.
Additional precautions to prevent transfer of Whirling Disease spores, New Zealand mudsnails, and
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          Option B:
                                           Equipment Care after Sampling
          amphibian chytrid fungus.
          Before visiting the site, consult the site dossier and determine if it is in an area where whirling disease,
          New Zealand mud snails, or chytrid fungus are known to exist. Contact the local State fishery biologist to
          confirm the existence or absence of these organisms.
          If the stream is listed as "positive" for any of the organisms, or no information is available, avoid using
          felt-soled wading boots, and, after sampling, disinfect all fish and benthos sampling gear and other
          equipment that came into contact with water or sediments (i.e., waders, boots, etc.) by one of the
          following procedures:
          Option A:
              1.   Soak gear in a 10% household bleach solution for at least 10 minutes, or wipe or spray on a 50%
                  household bleach solution and let stand for 5 minutes
              2.   Rinse with clean water (do not use stream water), and remove remaining debris
              3.   Place gear in a freezer overnight or soak in a 50% solution of Formula 409® antibacterial cleaner
                  for at least 10 minutes or soak gear in 120°F (49°C) water for at least 1 minute.
              4.   Dry gear in direct sunlight (at least 84 °F) for at least 4 hours.
              1.
                 Soak gear in a solution of Sparquat® (4-6 oz. per gallon of water) for at least 10 minutes
                 (Sparquat is especially effective at inactivating whirling disease spores).
             2.  Place gear in a freezer overnight or soak in 120°F (49°C) water for at least 1 min.
             3.  Dry gear in  direct sunlight (at least 84 °F) for at least 4 hours.
          2. Clean and dry other equipment prior to storage.
          Rinse coolers with water to clean off any dirt or debris on the outside and inside.
          Rinse periphyton sampling equipment with tap water at the base location.
          Make sure conductivity meter probes are rinsed with deionized water and stored moist.
          Rinse carboy and all beakers used to collect water chemistry samples three times with deionized water.
          Place beakers in a 1-gallon sealable plastic bag with a cube container for use at the next stream.
          Check nets for holes and repair or locate replacements.
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          3. Inventory equipment and supply needs and relay orders to the Field Logistics Coordinator.
          4. Remove GPS, multi-probe meter, and electrofishing unit from carrying cases and set up for
               predeparture checks and calibration. Examine the oxygen membranes for cracks, wrinkles, or
               bubbles. Replace if necessary, allowing sufficient time for equilibration.
          5. Recharge/replace batteries as necessary.
          6. Replenish fuel and oil; if a commercial car wash facility is available, thoroughly clean vehicle and boat
          (hot water pressurized rinse, no soap).
          2.3     Safety and Health
          Collection and analysis of samples can involve significant risks to personal safety and health. This
          section describes recommended training, communications, and safety considerations, safety
          equipment and facilities, and safety guidelines for field operations.
          2.3.1   General Considerations
          Important considerations related to field safety are presented in Table 2.3. Please follow your
          own agency's health and safety protocols, or refer to the Health and Safety Guidance for Field
          Sampling: National Rivers and Streams Assessment (available from the EPA Regional
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Coordinator) and Logistics of Ecological Sampling on Large Rivers (Flotemersch, et al. (editors)
2000). Additional sources of information regarding safety-related training include the American
Red Cross (1979), the National Institute for Occupational Safety and Health (1981), U.S. Coast
Guard (1987) and Ohio EPA (1990).
Field crew members should become familiar with the hazards involved with sampling equipment
and establish appropriate safety practices prior to using them. They must make sure all
equipment is in safe working condition. Personnel must consider and prepare for hazards
associated with the operation of motor vehicles, boats, winches, tools, and other incidental
equipment. Boat operators should meet any state requirements for boat operation and be
familiar with U.S. Coast Guard rules and regulations for safe boating contained in a pamphlet,
"Federal Requirements for Recreational Boats," available from a local U.S. Coast Guard Director
or Auxiliary or State Boating Official (U.S. Coast Guard, 1987). Life jackets must be worn by crew
members at all times on the water. All boats with motors must have fire extinguishers, boat
horns, life jackets or flotation cushions, and flares or communication devices. Boats should stay
in visual contact with each other, and should use 2-way radios to communicate.
Primary responsibility for safety while electrofishing rests with the crew leader. Electrofishing
units may deliver a fatal electrical shock, and should only be used by qualified, experienced
operators. Field crew members using electrofishing equipment must be insulated from the
water, boat, and electrodes via rubber boots and linesman gloves. All personnel should use
chest waders with nonslip soles and linesman gloves. DO NOT wear breathable waders while
electrofishing.  If waders become wet inside, stop fishing until they are thoroughly dry or use a
dry pair. Avoid contact with the anode and cathode at all times due to the potential shock
hazard. If you perspire heavily, wear polypropylene or some other wicking and insulating
clothing instead of cotton. If it is necessary for a crew member to reach into the water to pick up
a fish or something that has been dropped, do so only after the electrical current is off and the
anode is removed from the water. Do not resume electrofishing until all individuals are clear of
the electroshock hazard. Ensure that the backpack electrofishing equipment is equipped with a
45° tilt switch that  interrupts the current.  Do not make any modifications to the electrofishing
unit that  would hinder this safety switch. Avoid electrofishing near unprotected people, pets, or
livestock. Discontinue activity during thunderstorms or rain. Crew members should keep each
other in constant view or communication while electrofishing. For each site, know the location
of the nearest emergency care facility. Although the crew leader has authority,  each crew
member  has the responsibility to question and modify an operation or decline participation if it
is unsafe.
Table 2.3 General Health and Safety Considerations
                                  Recommended Training
  • First aid and cardiopulmonary resuscitation (CPR)
  • Vehicle safety (e.g., operation of 4-wheel drive vehicles)
  • Boating and water safety; Whitewater safety if applicable
  • Field safety (weather, personal safety, orienteering, site reconnaissance)
  • Equipment design, operation, and maintenance
  • Handling of chemicals and other hazardous materials
                                     Communications
  • Check-in schedule
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           • Sampling itinerary (vehicle used & description, time of departure & return, travel route)
           • Contacts for police, ambulance, hospitals, fire departments, search and rescue personnel
           • Emergency services available near each sampling site and base location
           • Cell (or satellite) phone and VHP radio if possible
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           • Field clothing and other protective gear including lifejackets for all crew members
           • Medical and personal information (allergies, personal health conditions)
           • Personal contacts (family, telephone numbers, etc.)
           • Physical exams and immunizations

         A communications plan to address safety and emergency situations is essential. All field
         personnel need to be fully aware of all lines of communication. Field personnel should have a
         daily check-in procedure for safety. An emergency communications plan should include contacts
         for police, ambulance, fire departments, hospitals, and search  and rescue personnel.
         Proper field clothing should be worn to prevent hypothermia, heat exhaustion, sunstroke,
         drowning, or other dangers. Field personnel must be able to swim, and personal flotation
         devices must be used. Chest waders made of rubberized or neoprene material must always be
         worn with a belt to prevent them from filling with water in case of a fall. A personal flotation
         device (PDF) and suitable footwear must be worn at all times while on board a boat.
         Many hazards lie out of sight in the bottoms of rivers and streams.  Broken glass or sharp pieces
         of metal embedded in the substrate can cause serious injury if care is not exercised when
         walking or working with the hands in such environments. Infectious agents and toxic substances
         that can be absorbed through the skin or inhaled may also be present in the water or sediment.
         Personnel who may be exposed to water known or suspected to contain human or animal
         wastes that carry causative agents or pathogens must be immunized against tetanus, hepatitis,
         typhoid fever, and polio. Biological wastes can also be a threat in the form of viruses, bacteria,
         rickettsia, fungi,  or parasites.

         2.3.2   Safety Equipment
         Appropriate safety apparel such as waders, linesman gloves, safety glasses, etc. must be
         available and used when necessary. First aid kits, fire extinguishers, and blankets must be readily
         available in the field. Cellular or satellite telephones and/or portable radios should be provided
         to field crews working in remote areas in case of an emergency. Supplies (e.g., clean water,
         antibacterial soap, ethyl alcohol) must be available for cleaning exposed body parts that may
         have been contaminated by pollutants in the water.

         2.3.3   Safety Guidelines for Field Operations
         General safety guidelines for field operations are presented in  Table 2.4. Personnel participating
         in field activities should be in sound physical condition and have a physical examination annually
         or in accordance with organizational requirements. All surface waters and sediments should be
         considered potential health hazards due to potential toxic substances or pathogens. Persons
         must become familiar with the health hazards associated with  using chemical fixing and/or
         preserving agents. Chemical wastes can be hazardous due to flammability, explosiveness,
         toxicity, causticity, or chemical reactivity. All chemical wastes must be discarded according to
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standardized health and hazards procedures (e.g., National Institute for Occupational Safety and
Health [1981]; U.S. EPA [1986]).

During the course of field research activities, field crews may observe violations of
environmental regulations, may discover improperly disposed hazardous materials, or may
observe or be involved with an accidental spill or release of hazardous materials. In such cases it
is important that the proper actions be taken and that field personnel do not expose themselves
to something harmful. The following guidelines should be applied:

    1.   First and foremost, protect the health and safety of all personnel. Take necessary steps
       to avoid injury or exposure to hazardous materials. If you have been trained to take
       action such as cleaning up a minor fuel spill during fueling of a boat, do it. However, you
       should always err on the side of personal safety.

    2.   Field personnel should never disturb or retrieve improperly disposed hazardous
       materials from the field to bring back to a facility for "disposal". To do  so may worsen
       the impact, incur personal liability for the crew members and/or their  respective
       organizations, cause personal injury, or cause unbudgeted expenditure of time and
       money for proper treatment and disposal of the material. Notify the appropriate
       authorities so they may properly respond to the incident.

    3.   For most environmental incidents, the following emergency telephone numbers should
       be provided to all field crews: State or Tribal department of environmental quality or
       protection, U.S. Coast Guard, and the U.S. EPA regional office. In the event of a major
       environmental incident, the National Response Center may need to be notified at 1-800-
       424-8802.

Table 2.4 General Safety Guidelines for Field Operations
        Two crew members must be present during all sample collection activities, and no one should be
        left alone while in the field. Boats should proceed together down the river.
        Use caution when sampling on foot in swift or deep water. Wear a suitable PFD and consider
        using a safety tether held by an assistant.
        Use extreme care walking on riprap. Rocks can shift unexpectedly and serious falls are possible.
        Field crew members using electrofishing equipment must be insulated from the water, boat, and
        electrodes via non-breathable waders and linesman gloves. Use chest waders with nonslip soles.
        Electrofishing units may deliver a fatal electrical shock, and should only be used by qualified,
        experienced operators.
        Do not attempt to collect samples from vertical or near vertical banks.
        Professional quality breathable waders with a belt are recommended for littoral sampling only,
        and at a safe distance from the electrofishing sampling. Neoprene boots are an alternative, but
        should have sturdy, puncture resistant soles.
        Exposure to water and sediments should be minimized as much as possible. Use gloves if
        necessary, and clean exposed body parts as soon as possible after contact.
        All electrical equipment must bear the approval seal of Underwriters Laboratories and must be
        properly grounded to protect against electric shock.
        Use heavy gloves when hands are used to agitate the  substrate during collection of benthic
        macroinvertebrate samples.
        Use appropriate protective equipment (e.g., gloves, safety glasses) when handling and using
        hazardous chemicals.
        Crews working in areas with poisonous snakes must check with the local Drug and Poison Control
        Center for recommendations on what should be done in case of a bite from a poisonous snake.
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                 Any person allergic to bee stings, other insect bites, or plants (i.e., poison ivy, oak, sumac, etc.)
                 must take proper precautions and have any needed medications handy.
                 Field personnel should also protect themselves against deer or wood ticks because of the
                 potential risk of acquiring pathogens that cause Rocky Mountain spotted fever and Lyme disease.
                 Field personnel should be familiar with the symptoms of hypothermia and know what to do in
                 case symptoms occur. Hypothermia can kill a person at temperatures much above freezing (up to
                 lOoC or 50oF) if he or she is exposed to wind or becomes wet.
                 Field personnel should be familiar with the symptoms of heat/sun stroke and be prepared to
                 move a suffering individual into cooler surroundings and hydrate immediately.
                 Handle and dispose of chemical wastes properly. Do not dispose any chemicals in the field.
          2.4    Forms (Paper or Electronic)
          Forms are the key to data collection and tracking for the NRSA 2013/14.Electronic forms have
          been developed as well as paper forms. These electronic forms should streamline data
          collection. Field crews will have the option of using paper or electronic forms.

          2.4.1   Field Forms
          Field forms are the primary documents where we record measures, observations, and collection
          information during the course of the field day. Additional information regarding specifics of data
          entry is contained in Section 1.6.

             •   Paper Field  Forms: A paper field form packet (wadeable or non-wadeable) for each site
                 will  be provided by the NARS IM Coordinator if you have elected to use paper field data
                 collection. You will need to add these forms to the site packet prior to going in the field.
                 After a site is sampled, the completed NRSA 2013/14 paper field forms  are checked for
                 completeness and organized sequentially into a Data Packet. The Data Packets from
                 several sites are batched together and sent every 1-2 weeks to the NARS IM Center and
                 accompanied by a Tracking form to track which data packets have been shipped. Extra
                 paper field forms will be provided to field crews to serve as backup copies in case of lost
                 forms or problems with electronic field forms.

             •   Electronic Field Forms: This form of data collection can be collected through 3
                 platforms: an iOS, Android or a Windows portable electronic device (tablets, phones).
                 This will require a field crew to download or install the developed Application (or "App")
                 onto the device. The field forms will be optimized for tablet devices. Once downloaded
 ^               and the App launched, the field forms will be split into sections or "form-lets" for easier
 ^               data entry.  It is important for a field crew to familiarize themselves with the App prior
 ^               to field sampling.
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 5j        Tracking forms describe the status and location of all samples collected during NRSA 2013/14.
 <        Field crew leaders will typically transmit these forms electronically (by emailing a fillable PDF
 ^        form) to the NARS IM Center at specified times and you will pack hard copies in shipping
 H        containers with the samples. See APPENDIX C: SHIPPING GUIDELINES for more information.
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 p           •   Site and Sample Status/Water Chemistry Lab Tracking: Transmitted within 24 hours of
 ^               sampling or visiting a site to report on the status of the site (e.g. sampleable or not), to
 §               record the Sample ID numbers, and to indicate the status of all samples collected at the
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       site (immediate shipment and batch shipments). This also serves as the tracking form for
       sample shipped to the WRS lab.

    •  Tracking - Packets: Accompany packets that are batched together from multiple sites
       and shipped every 1 or 2 weeks. These are sent to the NARS IM Center.

    •  Tracking -Samples: Accompany samples that are batched together from multiple sites
       and shipped every 1 or 2 weeks. Whenever batched samples are shipped to their
       designated lab for analysis, the appropriate tracking form, which lists the Sample ID
       numbers for all samples packed in a shipping container, is included in the shipping
       package and is also transmitted electronically to the  NARS IM Center. Separate forms
       exist for the tracking of frozen batched samples, non-chilled batched samples and whole
       fish samples

2.4.3  Equipment and Supplies

2.4.3.1 Request Form
Field Crews will  submit requests for field forms, labels and site kits via an electronic Request
Form. This form will be submitted to the NARS Information Management (IM) Coordinator who
will ensure that  the request reaches the appropriate entity. Crews must submit sampling
schedules at or before the time of submitting request forms. Crews should submit the Request
Form at least 2 weeks prior to their desired sampling date.

2.4.3.2 Base Kit
The Base Kit is comprised of the subset of durable equipment and supplies needed for NRSA
2013/14 sampling that is provided by USEPA through the Contract Field Logistics Coordinator.
Typically one Base Kit is provided to each Field Crew and contains some of the equipment that is
used throughout the field season. See APPENDIX A: EQUIPMENT & SUPPLIES for a list of the
items provided by USEPA in the Base Kit.  We anticipate that this equipment will be available for
use in future NRSA efforts.

2.4.3.3 Site Kit
A Site Kit contains the subset of consumable supplies (i.e., items used up during sampling or
requiring replacement after use) provided by USEPA through the Contract Field Logistics
Coordinator. The site kit will contain all the sample bottles necessary for sampling a single site. A
new Site Kit is provided for each site sampled. See APPENDIX A: EQUIPMENT & SUPPLIES for the          ^
consumable items that will be provided by USEPA.                                                  o-

2.4.3.4 Field Crew Supplied Items                                                                £
                                                                                              	i
The field crew will also supply particular items for the field sampling day. These might include            ^
supplies from  the previous NRSA, typical field equipment (like a GPS), or boat equipment. See            Q
APPENDIX A: EQUIPMENT & SUPPLIES for the items that the field crew will need to provide.              5
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          3      INITIAL SITE PROCEDURES
          When you arrive at a site, you must first confirm you are at the correct site, and then determine
          if the site meets the criteria for sampling and data collection activities (See Site Evaluation
          Guidelines EPA-841-B-12-008). Inspect the selected reach for appropriate access, safety, and
          general conditions. Decide whether the site is at base flow condition and not unduly influenced
          by rain events which could affect the representativeness of field data and samples. If you
          determine that the site can be sampled, lay out a defined reach within which all sampling and
          measurement activities are conducted.

          3.1    Site Verificatio n Activities

          3.1.1   Locating the X-Site
          River and stream sampling points were chosen using the National Hydrography Dataset (NHD),
          in particular NHD-Plus, following a systematic randomized selection process (Stevens and Olsen,
          2004). Each point is referred to as the "X-site." The "X-site" is the mid-point of the sampling
          reach, and it will determine the location and extent for the rest of the sampling reach. The
          latitude/longitude of the "X-site" is listed on the site spreadsheet that was distributed to each
          field crew leader. Table 3.1 provides the equipment and supplies needed for site verification.
          Note that the coordinates provided on the site spreadsheet may not be located in the middle of
          the stream or river; and in some cases, the coordinates  may be on dry land next to the stream or
          river.  In these cases, it is important for crews to locate the x-site at a point that is in the middle
          of the stream or river (e.g. midway between the left and right banks).  To do this, simply
          measure the distance between banks and move the point perpendicular to the nearest bank
          until it is half-way between the left and right banks. Record these coordinates as the x-site on
          the Verification Form. If the site falls on dry land near a  stream, move the X site to the nearest
          blue line.  Record the coordinates as the x-site on the Verification Form.
          Table 3.1 Equipment and Supplies: for Site Verification

          For locating and      Sampling permit and landowner access (if required)
          verifying site         f\e\d Operations Manual and laminated Quick Reference Guide
                             Site dossier, including access information, site spreadsheet with map coordinates,
                               street and/or topographic maps with "X-site" marked
                             NRSA 2013-14 Community Fact Sheet
                             GPS unit (preferably one capable of recording waypoints) with manual, reference
                               card,  extra battery pack
                             Surveyor's flagging tape (to mark transects)
                             Laser rangefinder
 u-,                           50 m or 100 m measuring tape with reel (if not using rangefinder)
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 g        For recording        Clipboard
 S        measurements       #2 pencNs
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3.1.2   Determining the Sampling Status of a Stream
After you confirm the location of the X-site, evaluate the stream reach surrounding the X-site
and classify the stream into one of three major sampling status categories: sampleable, non-
sampleable, or no access (Table 3.2). The primary distinction between "Sampleable" and "Non-
Sampleable" streams is based on the presence of a defined stream channel, water content
during base flow, and adequate access to the site.
There must be greater than 50% water throughout the channel reach. If the channel is dry at
the X-site, determine if water is present within 75 m  upstream and downstream of the X-site. If
there are isolated pools of water within the reach that equal greater than 50% of the reach
length, proceed to sample using the modified procedures outlined in Section 3.1.1. Do not drop
the site  if it is dry at the X site, as long as there is greater than 50% water throughout the
channel. If less than 50% of the reach has water, classify the site as "Dry-visited" on the
Verification Form. NOTE: Do not "slide" the reach (Section 3.2.1) for the sole purpose of
obtaining more water to sample (e.g., the downstream portion of the reach has water, but the
upstream portion does not).
Record the sampling status and pertinent site verification information on the Verification  Form
(Figure 3.1). If the site is non-sampleable or inaccessible, no further sampling activities are
conducted. Replace the site with the first oversample site on the state list within the same
category based on Strahler order and whether it is a  2008/09 resample site (Section 1.3).
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 a.
                  Site ID:
                                                       NRSA 2013/14 VERIFICATION (Front)
                                                                 Visit: O1   O2  Date:         /
                  Site Name:
                                                                           State of Site Location:
                  STREAM/RIVER VERIFICATION INFORMATION
                    Stream/River verified by (Mark all that apply):  O GPS
                     O Other (Describe Here):
                                                         O Local Contact    O Signs
O Roads     O Topo. Map
                       Coordinates
                    GPS
                          Decimal Degrees
                                           Latitude
                                                                   Longitude
                                                                                  Type of GPS Fix  Elevation at transect A
                                                                                                Q2D  Q3D
                                Location:   O *-SUe (wadeable)   O Transect A (non-wadeable)
                                                                                                                    Oft   Om
                  DID YOU SAMPLE THIS SITE?
                   O YES  If Yes, check one below
                     SAMPLEABLE (Choose method used)
                   O Wadeable - Continuous water, greater than 50% wadeable
                   O Boatablo
                   O Partial-Sampled by wading (>50% of reach sampled). Explain below,
                   O Partial -Sampled by boat(>5G% of reach sampled). Explain below.
                   O Wadeablelnterrupted -Not eorrtlnuous water along reach
                   O Scalable Interrupted -Not continuous water along reach
                   O Altered - Stream.'River Channel Present but differs from Map
                   ADDITIONAL SITE CHARACTERISTICS
                   O Tidnlly Influenced   O Blnckwater   Q Not Applicable
                                                                   O N0  If No, check one below
                                                                    NON-SAMPLEABLE-PERMANENT
                                                                  O Dry-Visited
                                                                  O Dry-Not visited
                                                                  O Wetland (No Definable Channel)
                                                                  O Map Error (No evidence channel/walerbody ever present)
                                                                  O Impounded (>7day residence time)
                                                                  O Tidal (exceeds salinity threshold)
                                                                    NON-SAMPLEABLE-TEMPORARY
                                                                  O Other (explain In comments)
                                                                  O Not boatable - Need a different crew - Reschedule for this year
                                                                  O Not wadeable - Need a different crew - Reschedule for this year
                                                                    NO ACCESS
                                                                  O Other (Explain In comments)
                                                                  O Access Permission Denied
                                                                  O Permanently Inaccessible (Unable/UnsafetoReachStte)
                                                                  O Temporarily Inaccessible-Fire, etc. - Reschedulefornextyear
                  GENERAL COMMENTS
                  DIRECTIONS TO SITE
                        04/08/2013   2013 Verification
                                                                                                                5409334985
Figure 3.1 Verification  Form (front)
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             Wadeable
Table 3.2 Procedure: Site Verification
  1.   Find the stream/river location in the field corresponding to the X-site coordinates. Record the
      routes taken and other directions on the Verification Form so that others can visit the same
      location in the future. If the site is non-wadeable, locate public or private launch sites.
  2.   Use a GPS receiver to confirm the latitude and longitude at the X-site with the coordinates
      provided for the site (datum = NAD 83). Record these on the Verification Form.
  3.   Use all available means to insure you are at the correct stream/river as marked on the map,
      including 1:24,000 United States Geological Society (USGS) maps, topographic landmarks, road
      maps, signs, local contacts, etc.
  4.   Scan  the channel upstream and downstream from the X-site, decide if the site is sampleable, and
      mark the appropriate bubble on the Verification Form.
  5.   If the channel is dry at the X-site, determine if water is present within 75 m upstream and
      downstream of the X-site. Assign one of the following sampling status categories to the stream.
      Record the category on the Verification Form.
                                     Sampleable Categories
  Wadeable: Continuous water, sampled by wading.
  Boatable: Continuous water, too deep to sample by wading.
  Partial wadeable: Sampled by wading (>50% of reach sampled).
  Partial boatable: Sampled by boat (>50% of reach sampled).
  Wadeable Interrupted: not continuous water along reach, >50% water in reach.
  Boatable Interrupted: not continuous water along reach, >50% water in  reach.
  Altered Channel: Stream/river channel present but differs from map.
                                  Non-Sampleable Categories
PERMANENT
  Dry Channel: Less than 50% water within the reach. Record as "Dry-Visited." If site was determined to
    be dry (or otherwise non-perennial) from another source and/or field verified before the actual
    sampling visit, record as "Dry-Not visited".
  Wetland: Standing water present, but no definable stream channel. If wetland is surrounding a stream
    channel, define the site as Target but restrict sampling to the stream channel.
  Map Error: No evidence that a water body or stream channel was ever  present at the X-site.
  Impounded stream: Stream is submerged under a lake or pond due to manmade or natural (e.g., beaver
    dam) impoundments. If the impounded stream is still wadeable, record it as "Altered" and sample.
  Other: Examples would include underground pipelines, or a non-target canal. A sampling site must meet
    both of the following criteria to be classified as a non-target canal:
  The channel is constructed where no natural channel has ever existed.
  The sole purpose/usage of the reach is to transfer water. There are no other uses of the waterbody by
    humans (e.g., fishing, swimming, and boating).
TEMPORARY
  Not Boatable: need a different crew.
  Not Wadeable: need a different crew.
  Other: The site could not be sampled on that particular day, but is still a target site. Examples might
  include a recent precipitation event that has caused unrepresentative conditions.
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                                          No Access to Site Categories
            Access Permission Denied: You are denied access to the site by the landowner.
            Permanently Inaccessible: Site is unlikely to be sampled by anyone due to physical barriers that prevent
            access to the site (e.g., cliffs).
            Temporarily Inaccessible: Site cannot be reached due to barriers that may not be present at a future
            date (e.g. forest fire, high water, road temporarily closed, unsafe weather conditions).

            6.  Do not sample non-target or "Non-sampleable" or "No Access" sites. Fill in the "NO" bubble for
               "Did you sample this site?" and mark the appropriate bubble in the "Non-Sampleable" or "No
               Access" section of the Verification Form; provide a detailed explanation in comments section.
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3.1.3   Elevation at Transect A
Record elevation at Transect A using your GPS device. To record this information, record the
elevation holding the GPS at approximately 3 feet above the surface of the water. Ensure that
the numbers are properly recorded from Transect A on the Site Verification form.

3.1.4   Sampling During or After Rain Events
Avoid sampling during high flow rainstorm events. Use your best professional judgement to
determine if the stream has risen above baseflow during this recent rain event. It is often unsafe
to be in the water during such times. In addition, biological and chemical conditions during such
episodes are often quite different from those during baseflow. On the other hand, sampling
cannot be restricted to only strict baseflow conditions. It would be next to impossible to define
"strict baseflow" with any certainty at  an unstudied site. Such a restriction would also greatly
shorten the  index period  when sampling activities can be conducted. Thus, some compromise is
necessary regarding whether to sample a given stream because of storm events. To a great
extent, this decision is based on the judgment of the field crew. Some guidelines to help make
this decision are presented in Table 3.3. The major indicator of the influence of storm events
will be the condition  of the stream itself. If you decide a site is unduly influenced by a storm
event, do not sample the site that day.
Table 3.3 Guidelines to Determine the Influence of Rain Events
  •   If it is running at bank full discharge or the water seems much more turbid than typical for the class
     of stream do not sample it that day.
  •   Do not sample that day if it is unsafe to be in the water.
  •   Keep an eye on the weather reports and rainfall patterns. Do not sample a stream during periods of
     prolonged heavy rains.
  •   If the stream seems to be close to normal summer flows, and does not seem to be unduly
     influenced by storm events, sample it even if it has recently rained  or is raining.

3.1.5   Site Photographs
Taking site photographs is an optional  activity, but should be considered if the site has unusual
natural or manmade features associated with it. If you do take photographs with a digital
camera at a  site, date stamp the photograph and include the site ID. Alternatively, start the
sequence with one photograph of an 8.5 x 11 inch piece of paper with the site ID, waterbody
name, and date printed in large, thick letters. After the photo of the site ID information, take at
least two photographs at the X-site, one in the upstream direction and one downstream. Take
any additional photos you find interesting after these first three pictures. Keep a log of your
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photographs and briefly describe each one. Photographs can be uploaded to the NARS Sharefile
site.

3.2     Laying out the sampling reach

Many of the biological and habitat structure measures require sampling a certain length of a
stream to get a representative picture of the ecological community. A length of 40 times the
wetted width is necessary to characterize the habitat and several biotic assemblages
associated with the sampling reach. Establish the sampling reach about the X-site using the
procedures described in Table 3.4 (wadeable sites).

When you arrive at the site, scout the sampling reach to make sure it is clear of obstacles that
would prohibit sampling and data collection activities. Record the channel width used to
determine the reach length, and the sampling reach length upstream and downstream of the X-
site on the Verification Form (back) as shown in Figure 3.2. Figure 3.3 illustrates the principal
features of the established sampling reach for wadeable sites, including the location of 11 cross-
section transects used for collecting samples and physical habitat measurements. The figures
also show the specific sampling stations on each transect for collection of periphyton, and
benthic macroinvertebrate samples.

Before leaving the stream, complete a rough sketch map of the stream reach you sampled on
page 2 of the Verification  Form (Figure 3.2). In addition to any other  interesting features that
should be marked  on the map, note any landmarks/directions that can be used to find the X-site
for future visits.

Table 3.4 Procedure: Laying Out the Sampling Reach at Wadeable Sites

  1.   Locate the X-site using the coordinates provided for the site.
  2.   Use a surveyor's rod, tape measure, or laser range finder to determine the wetted width of the
      channel at 5 places of "typical" width within approximately 5 channel widths upstream and
      downstream from the X-site. Average the 5 readings together and round to the nearest 1 m. If the
      average width is <4 m, use 150 m as a minimum reach length. If the average width is >100 m, use 4
      km as a maximum reach length. Record this width on page 2 of the Verification Form.
  3.   For channels with "interrupted flow", estimate the width based on the unvegetated width of the
      channel (again, with a 150 m minimum and 4 km maximum).
  4.   Check the condition of the stream about the X-site by having one crew member go upstream and
      one downstream. Each person proceeds until they can see the stream to a distance of 20 times the
      average channel width (equal to one-half the sampling reach length) determined in Step 2.
  5.   Determine if the reach needs to be adjusted about the X-site due to confluences with higher order
      streams (downstream), transitions into lower order streams (upstream), impoundments (lakes,
      reservoirs, ponds), physical barriers (e.g., falls, cliffs), or because of access restrictions to a portion
      of the initially determined sampling reach. Refer to Section 3.2.1.
  6.   Starting at the X-site (or the new midpoint of the reach if it had to be adjusted as described in Step          ^
      4), measure a distance of 20 channel widths down one side of the stream using a GPS unit, laser             ^
      rangefinder, or tape measure. Be careful not to "cut corners". Enter the channel to make                   ^
      measurements only when necessary to avoid disturbing the stream channel prior to sampling               £j
      activities. This endpoint is the downstream end of the reach, and is flagged as Transect "A".                 O
  7.   At Transect A, use the seconds display on a digital watch to select the initial sampling station for             ^]
      standard transect samples: l-3="Left", 4-6="Center", 7-9=Right. Mark "L", "C", or "R" on the                t
      transect flagging; the 3 potential collection points are roughly equivalent to 25%, 50%, and  75% of           -j
      the channel width, respectively. Note that left and right sides of the stream are determined while            p
      you  are facing downstream. It is at these locations that you will collect benthic macroinvertebrate           ^


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                and periphyton samples.
            8.  Measure 1/10 of the required reach length upstream from transect A. Flag this spot as transect B.
                Assign the sampling station systematically after the first random selection following the repeating
                pattern  L, C, R as you  move upstream (Figure 3.3).
            9.  Proceed upstream with the tape measure and flag the positions of 9 additional transects (labeled
                "C" through "K" as you move upstream) at intervals equal to 1/10 of the reach length. Continue to
                assign the sampling stations systematically.
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                                       NRSA 2013/14 VERIFICATION (Back)


                                                Visit: 0 1   O 2  Date:         1         /
     STREAM/RIVER REACH DETERMINATION
Channel Width
Used to Define
Reach(m)
                         DISTANCE (m) FROM X-SITE
                   Upstream Length:
Downstream Length:
                 Total Reach
                 Length Intended (m):
                                                                        Comment:
     SKETCH MAP
     Arrow Indicates North; Mark site L=Launch X=lndex T- Take Out
     NOTE: If an outline map Is attached here, use a continuous strip of clear tape across the top edge. You can also attach a separate sheet with the
     outline map on it. For beatable sites you can attach topo map with reach, X-slte and transect locations marked.
     PERSONNEL
    Crew Leader


    Fish Taxonomist


    Name:


    Name
                     Name:


                     Name:


                     Name:


                     Name:
          04/08/2013  2013 Verification
                                                                                             4426334983
Figure 3.2 Verification Form (back)
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                                                            SAMPLING POINTS
                                                            • l_=l_eft C=Center R=Right
                                                            • First point (transect A)
                                                             determined at random
                                                            • Subsequent points assigned in
                                                             order L, C, R
    Distance between transects=4 times
    mean wetted width at X -site
                     Total reach length=40 times mean wetted width at X -site (minimum=1 50
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Figure 3.3 Sampling Reach Features for a Wadeable Site

3.2.1   Sliding the Reach

There are some conditions that may require sliding the reach about the X-site (i.e., the X-site is
no longer located at the midpoint of the reach) to avoid features we do not wish to or physically
cannot sample across. Reasons for sliding the reach include:

    •   Landowner access;

    •   Confluence with higher order waterbody;

    •   Impoundment;

    •   Impassable barrier.

Sliding the reach involves noting the distance of the barrier, confluence, or other restriction
from the X-site, and flagging the restriction as the endpoint of the reach. Add the distance to the
other end of the reach, such that the total reach length remains the same, but it is no longer
centered about the X-site. Table 3.5 describes when you should and  should not slide the
sampling reach.

Table 3.5 Procedure: Sliding the Sampling Reach

  1.   Slide the reach if you run into an impoundment (lake, pond, or reservoir), so that the lake/stream
      confluence is at one end.
  2.   Slide the reach if you run into an impassible barrier (e.g., waterfall, cliff, navigation dam) so that
      the barrier is at one end.
  3.   Slide the reach if you run into a confluence (another stream meeting the water-body you are
      sampling) with a higher Strahler Order.
  4.   When  you are denied access permission to a portion of the reach, you can slide the reach to make
      it entirely accessible; use the point of access restriction as the endpoint of the reach.
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  5.   Note the distance of the barrier, confluence, or other restriction from the X-site, and flag the
      restriction as the endpoint of the reach. Add the distance to the other end of the reach, so the total
      reach length remains the same, but it is no longer centered about the X-site.
  6.   Do not slide the reach so that the X-site falls outside of the reach boundaries.
  7.   Do not proceed upstream into a lower order stream or downstream into a higher order stream
      when laying out the stream reach (order is based on 1:100,000 scale maps).
  8.   Do not slide a reach to avoid man-made obstacles such as bridges, culverts, rip-rap, or
      channelization. These represent important features and effects to study.
  9.   Do not slide a reach to gain more water to sample if the flow is interrupted.
  10. Do not slide a reach to gain better habitat for benthos or fish.

3.3     Modifying Sample Protocols for High or Low Flows

3.3.1   Streams with Interrupted Flow
You cannot collect the full complement of field data and samples from streams that are
categorized as "Interrupted" (Table 3.6). Note that no data should be collected from streams
that are completely "Dry" as defined in Table 3.6. Interrupted streams will have some cross-
sections amenable to biological sampling and habitat measurements and some that are not. To
be considered target, streams must have greater than 50% water in the reach length within the
channel (can be isolated pools).  Modified procedures for interrupted streams are presented in
Table 3.6. Samples for water chemistry (Section 0) will be collected at the X-site (even if the
reach has been adjusted by "sliding" it). If the X-site is dry and there is water elsewhere in the
sample reach, collect the sample from  a location having water with a surface area >1 m2 and a
depth >10 cm.
Collect data for the physical habitat indicator along the entire sample reach from interrupted
streams, regardless of the amount of water present at the transects. Obtain depth
measurements along the deepest part of the channel (the "thalweg") along the entire sampling
reach to provide a record of the "water" status of the stream for future comparisons (e.g., the
percent of length with intermittent pools or no water). Other measurements associated with
characterizing riparian condition, substrate type, etc., are useful to help infer conditions in the
stream when water is flowing.
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          Table 3.6 Reach Layout Modifications for Interrupted Streams
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          Streams with less than 50% of reach length containing water (not necessarily continuous) are considered
          dry and are not sampled.
          If more than 50% of the channel has water and if the X-site is dry but there is flowing water or a pool of
          water having a surface area > 1 m2 and a depth > 10 cm somewhere along the defined sampling reach, take
          the water sample at the  pool or flowing water location that is nearest to the X-site. Note that the sample
          wasn't collected at the X-site and where on the reach the sample was collected on the field data form.
          Do not collect a water sample  if there is no acceptable location within the sampling reach. Record a "K"
          flag for the water chemistry sample on the sample collection form and explain why the sample was not
          collected in the comments section of the form.
                             Physical Habitat, Periphyton and Benthic Macroinvertebrates
          Obtain a complete thalweg profile for the entire reach. At points where the channel is dry, record depth as
          0 cm and wetted width as 0 m.
          At each of the transects (cross-sections), sample the stream depending on flow status:
          DRY CHANNEL: No surface water anywhere in cross-section; collect all physical habitat data. Use the
          unvegetated area of the channel to determine the channel width and the subsequent location of substrate
          sampling points. Record the wetted width as 0 m. Record substrate data at the sampling points  located in
          the unvegetated, but dry, channel. Do not collect periphyton or benthic macroinvertebrates from this
          transect.
          DAMP CHANNEL: No flowing water at transect, only puddles of water < 10 cm deep; collect all physical
          habitat data. Do not collect periphyton or benthic macroinvertebrates from this transect.
          WATER PRESENT: Transect has flow or pools > 10 cm deep; collect all data and measurements for physical
          habitat, periphyton, benthic macroinvertebrate, and fish indicators, using standard procedures.
3.3.2   Braided Rivers and Streams
Depending upon the geographic area and/or the time of the sampling visit, you may encounter a
stream having "braided" channels, which are characterized by numerous sub-channels that are
generally small and short, often with no obvious dominant channel. If you encounter a braided
stream, establish the sampling reach using the procedures presented in Table 3.7. Figuring the
mean width of extensively braided rivers and streams for purposes of setting up the sampling
reach length is challenging. For braided channels, measure the mean width and bankfull width
as defined in the physical habitat protocols (Section Error! Reference source not found.). For
relatively small streams (mean bankfull width <15 m) the sampling reach is defined as 40 times
the mean bankfull  width. For larger streams (>15 m),  sum the actual wetted width of all the
braids and use that as the width for calculating the 40 channel width reach length. If there is any
question regarding an appropriate reach length for the braided system, it is better to
overestimate. Make  detailed notes and sketches on the Verification Form (Figure 3.1 and Figure
3.2) about what you  did. It is important to remember that the purpose of the 40 channel width
reach length is to sample enough streams to incorporate the variability in habitat types.
Generally, the objective is to sample a long enough stretch of a stream to include 2 to 3
meander cycles (about 6 pool riffle habitat sequences). In the case of braided systems, the
objective of this protocol modification is to avoid sampling an excessively long stretch of stream.
In a braided system where there is a 100 m wide active channel (giving a 4 km reach length
based on the standard procedure)  and only 10 m of wetted width  (say five, 2 m wide braids), a
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400 m long sample reach length is likely to be sufficient, especially if the system has fairly
homogenous habitat throughout its length.

Table 3.7 Procedure: Modifications for Braided Rivers and Streams

  1.   Estimate the mean width as the bankfull channel width as defined in the physical habitat protocol.
      •   If the mean width is <15 m, set up a 40 x channel width sample reach in the normal manner,
          using the mean bankfull width for your calculations.
      •   If >15 m, sum up the actual wetted width of all the braids and use that as the width for
          calculating the 40 x channel width reach length. Remember the minimum reach length is
          always 150 m.
      •   If the reach length seems too short for the system in question, set up a longer sample  reach,
          taking into consideration that the objective is to sample a long enough stretch of a stream to
          include at least 2 to 3 meander cycles (about 6 pool riffle habitat sequences).
  2.   Make detailed notes and sketches on the Verification Form about what you did.
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         4      WATER CHEMISTRY / CHLOROPHYLL-a SAMPLE COLLECTION
                AND PRESERVATION

         4.1    In Situ Measurements of Dissolved Oxygen, pH, Temperature, and
                Conductivity

         4.1.1   Summary of Method
         Measure dissolved oxygen (DO), pH, temperature, and conductivity using a calibrated multi-
         parameter water quality meter (or sonde). Take the measurements mid-channel at the X-site.
         Take the readings at 0.5 m depth.  Measure the site depth accurately before taking the
         measurements. If the depth at the x-site is less than 1 meter, take the measurements at mid-
         depth. Take care to avoid the probe contacting bottom sediments, as the instruments are
         delicate. Record the measurements on the Field Measurement Form, as seen in Figure 4.1.

         4.1.2   Equipment and Supplies
         Table 4.1 provides the equipment and supplies needed to measure dissolved oxygen, pH,
         temperature, and conductivity.
         Table 4.1 Equipment and Supplies: DO, pH, Temperature, and Conductivity

         For taking measurements and        Multi-parameter water quality meter with pH, DO, temperature,
         calibrating the water quality meter    and conductivity probes.
 °                                       Extra batteries

                                          De-ionized (Dl) and tap water
                                          Calibration cups and standards
                                          Barometer or elevation chart to use for calibration
         For recording measurements
Field Measurement Form
                                          Pencils (for data forms)
 O
 b
 ^       4.1.2.1  Multi-Probe Sonde
 	i
 u       Dissolved Oxygen Meter
 LJJ
 Q-       Calibrate the DO meter prior to each sampling event. We recommend that the probe be
 <       calibrated in the field against an atmospheric standard (ambient air saturated with water) prior
 CO
 re       to launching the boat. Follow your manufacturer's guidelines for calibration of the DO probe.
 >       pH Meter
 Q_
 O       Calibrate the pH meter prior to each sampling event. Calibrate the meter in accordance with the
 O       manufacturer's instructions and with the crew agency's existing Standard Operating Procedures
 5       (SOP).
 >       Temperature Meter
 —       You must check the accuracy of the sensor against a thermometer that is traceable to the
 LJJ       National Institute of Standards (NIST) at least once per sampling season. The entire temperature
 u       range encountered in the NRSA should be incorporated in the testing procedure and a record of
 LJJ       test results kept on file.
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Conductivity Meter

Calibrate the conductivity meter prior to each sampling event. Calibrate the meter in
accordance with the manufacturer's instructions. The entire conductivity range encountered in
the NRSA should be incorporated in the testing procedure and a record of test results kept on
file.

4.1.3   Sampling Procedure

Table 4.2 presents step by step procedures for measuring dissolved oxygen, pH, temperature,
and conductivity.

Table 4.2 Procedure: Temperature, pH, Conductivity and Dissolved Oxygen

  1.   Check Meter and probes and calibrate according to manufacturer's specifications.
  2.   Check the calibration against the QCS solution for pH and conductivity and record the results on the
      field sheet as the QCS Measured value.  This should be done at least once a week.
  3.   Record the true value of the QCS solution from the stock solution container on the field sheet as
      QCS True.
  4.   Samples are taken mid-channel, at the X site, at a depth of 0.5 meters or at a mid-depth if less than
      1 meter deep.
  5.   Lower the sonde in the water and measure DO, pH, temperature, and conductivity at 0.5 m depth
      (or at mid-depth if less than 1 meter deep).
  6.   Record the measurements on the Field Measurement Form                                            z
                                                                                                    O
  7.   Flag any measurements that the crew feels needs further comment or when a measurement                p
      cannot be made.                                                                                >
                                                                                                    cc.
                                                                                                    LJJ
                                                                                                    l/l
                                                                                                    LJJ
                                                                                                    CC
                                                                                                    CL
                                                                                                    0
                                                                                                    b
                                                                                                    _
                                                                                                    8
                                                                                                    CL
                                                                                                    O
                                                                                                    cc
                                                                                                    O
                                                                                                    U
                                                                                                    U
                                                                                                    QC
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Field Operations Manual
            Wadeable
 O

 £
 QC
 LLJ
 l/l
 LLJ
 QC
 a.
 Q
 g
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 8
 CL
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 C£
 2
 u
_ NRSA 2013/14 FIELD MEAS
Site ID:
JREMENT Rtvl™Mby(lrMi=i •
Date: / /
CALIBRATION INFORMATION
Instrument manufacturer and model:
Instrument ID number:
TEMPERATURE
DO
pH
CONDUCTIVITY
Thermometer
Reading C:C) Sensor Reading (CC) Comments

o_ Barometnc
Elevation Pressure (mm Hg) Calibration

Operator:


i/alue DisplayedValue Flag
O mg/L O moA
Gal STD 1 Description CalSTDIValue Cal.STD 2 Description Cal STD2Value
i i ii i i
Calibration Verified with Quality Control Sample (QCSi
QCS Description

1 1 11 1 1
GCSTrue QCSMeasured Flag

Cal STD 1 Description CalSTDIValue Cal STD 2 Descripiion Cal STD 2 Value


Calibration Verified with Quality Control Sample IOCS)
OCS Description DCS True (uSfcm @25"C) OCSMeasured(uSA:m@25'C) Flag

Flag Comments







Flag codes: K - No measurement or observation mode: U = Suspect measurement or observation : F1 F2 etc = rmsc lags assigned by field crew Explain all Hags in comment
sections
FIELD MEASUREMENT O X-Slte iwadeable) O Transect A (non-wadeable) O OtherTransect:

Time of Day (hh:mm) ;
DO(mg/L)XX.X
Temp. ("QXX.X
pH XX XX
Cord (uS/cm>XXX
Corrected to 25'C? QV ON
H 04/08/2013 2013 Field Measurement
tBrnnrpents






2045648267 |
          Figure 4.1 Field Measurement Form
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 QC
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                                                                               Wadeable
4.2    Water Chemistry Samples
4.2.1  Summary of Method
The water chemistry samples will be analyzed for total phosphorus (TP), total nitrogen (TN),
total ammonium (NH4), nitrate (NO3), basic anions, cations, total suspended solids (TSS),
turbidity, acid neutralizing capacity (ANC, alkalinity), dissolved organic carbon (DOC), and total
organic carbon (TOC). All field crews must collect a grab sample in one 4-L cube container and in
one 2-L amber Nalgene bottle from the X site at the midpoint of the reach. Store all samples on
ice in a closed cooler.
4.2.2  Equipment and Supplies
Table 4.3 provides the equipment and supplies needed to collect water samples at the x site.
Record the Water Sample Collection and Preservation data on the Sample Collection Form, as
seen in Figure 4.2.
Table 4.3 Equipment and Supplies: Water Chemistry Sample Collection and Preservation
For collecting samples
                       Nitrile gloves
                       4-L cube container
                       2-L amber Nalgene bottle
                       3-L Nalgene beaker
                       Cooler with ice
                       Dl water (for cleaning beaker between sites)
                       Field Operations Manual and laminated Quick Reference Guide
                                                                                                O
                                                                                                LJJ
                                                                                                l/l
                                                                                                LJJ
                                                                                                QC
                                                                                                CL
                                                                                                 .
                                                                                                0
                                                                                                b
                                                                                                8
For recording
measurements
Sample Collection Form
Water Chemistry sample label with pre-printed Sample ID
Clear tape strips
Field Measurement Form
Pencils (for data forms)
Fine tipped indelible markers
4.2.3  Water Chemistry and Chlorophyll-a Sampling Procedure
Table 4.4 presents step-by-step procedures for collecting water chemistry samples at wadeable
sites.
                                                                                                CL
                                                                                                O
                                                                                                O
                                                                                                I
                                                                                                U
                                                                                                U
                                                                                                QC
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          Table 4.4 Procedure: Water Chemistry and Chlorophyll-a Sample Collection (Wadeable Sites)

 LJJ
 l/l
 LJJ
 CC
 CL
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 8
 CL
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            1.
            2.

            3.

            4.
            5.


            6.


            7.
      Fill out the pertinent information (Site ID and date) on the water chemistry label and affix the label
      to the cube container. Completely cover the label with clear tape.
      Collect the water samples from the X-site in a flowing portion near the middle of the stream. Be
      sure to collect the water samples prior to any disturbance of the stream upstream of the X-site.
      Put on nitrile gloves. Make sure not to handle sunscreen or other chemical contaminants until after
      the sample is collected.
      Rinse the 3-L Nalgene beaker three times with  water, and discard the rinse downstream.
      Remove the cube container lid and expand the cube container by pulling out the sides. NOTE: DO
      NOT BLOW into the cube container or place your fingers inside the opening to expand it, this will
      cause  contamination.
      Fill the 3-liter beaker with water and slowly pour 30 - 50 mL into the cube container. Cap the cube
      container and rotate so that the water contacts all the surfaces. Discard the water downstream.
      Repeat this rinsing procedure 2 more times.
      Fill the beaker with water and pour into the cube container. Repeat as necessary to fill the cube
      container. Let the weight of the water expand the cube  container. Pour the water slowly as the
      cube container expands. Fill the cube container to at least three-fourths of its maximum volume.
      Rinse the cube  container lid with water. Eliminate any air space from the cube container, and cap it
      tightly. Make sure the cap is tightly sealed and  not on at an angle.
Chlorophyll-a
  8.   Fill the 3-liter beaker with water and slowly pour 30 - 50 mL into the 2 L amber Nalgene bottle. Cap
      the bottle and rotate so that the water contacts all the surfaces. Discard the water  downstream.
      Repeat this rinsing procedure 2 more times.
      Fill the beaker with water and pour into the 2 L amber Nalgene bottle, filling the bottle. Cap the
      bottle tightly. This sample will be filtered later and the bottle will be reused at future sites,
      therefore it is not necessary to label this bottle.


Place the cube container and Nalgene bottle in a cooler (on ice or water) and shut the lid. If a cooler is
not available, place the cube container in an opaque garbage bag and immerse it in the stream.
Record the Sample ID on the Sample Collection Form along with the pertinent stream information
(stream name,  ID, date,  etc.). Note anything that could influence sample chemistry (heavy rain, potential
contaminants)  in the Comments section. If the sample was collected at the X-site, darken  the X-site
bubble in the "STATION COLLECTED" field. If you had to move to another part of the reach to  collect the
sample, place the letter of the nearest transect  in the "STATION  COLLECTED" field. Record more detailed
reasons and/or information in the Comments section.
            9.
 U
 QC
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Field Operations Manual

            Wadeable
g NRSA 2013/14 SAMPLE
Site ID:
COLLECTION (Front) R™I»HIIIY
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          National Rivers and Streams Assessment 2013/14                          Field Operations Manual
          Version 1.0, May 2013                                                            Wadeable


          5      MICROCYSTINS
          Microcystis is a microscopic organism that is found naturally at low concentrations in freshwater
          systems. Under optimal conditions (such as high light and calm weather, usually in summer),
          Microcystis occasionally forms a bloom, or dense aggregation of cells, that floats on the surface
          of the water forming a thick layer or "mat." At higher concentrations, Microcystis blooms are so
          dense that they resemble bright green  paint that has been spilled in the water. These blooms
          potentially affect water quality as well  as human health (Microcystis produces microcystin, a
          potent liver toxin) and natural resources. Decomposition of large blooms can lower the
          concentration of dissolved oxygen in the water, resulting in hypoxia (low oxygen) or anoxia (no
          oxygen). Sometimes, this results in fish kills. The blooms can also be unsightly, often floating at
          the surface in a layer of decaying, odiferous, gelatinous scum.
          Although the likelihood of people being affected by a Microcystis bloom is low, minor skin
          irritation can occur with contact, and gastrointestinal discomfort can also occur if water from a
          bloom is ingested. People recreationally exposed (e.g., swimmers or personal watercraft
          operators) to microcystins have also reported minor skin irritation. Health problems may occur
          in animals if they are chronically exposed to fresh water with Microcystis present. Just as
          livestock and domestic animals can be  poisoned by drinking contaminated water, fish and bird
          mortalities have been reported in waterbodies with persistent Microcystis blooms.

          5.1     Summary of Method
          The microcystin (algal toxin) sample is a grab sample taken from the x site. All field crews must
          collect a grab sample using the 3 L beaker to fill a 500ml bottle. Collect this sample after the in
          situ measurements and water chemistry samples are collected. Store all samples on ice in a
          closed cooler.

          5.2     Equipment and Supplies
          Table 5.1 provides the equipment and supplies needed to collect microcystin sample at the
          index site. Record the Water Sample Collection and Preservation data on the Sample Collection
          Form, as seen in Error! Reference source not found..
          Table 5.1 Equipment and Supplies: Microcystin

          For collecting samples    Nitrile gloves
                                3-L Nalgene beaker
                                500ml Nalgene bottle
                                Cooler with ice
                                Field Operations Manual and laminated Quick Reference Guide

          For recording            Sample Collection Form
          measurements          Microcystin sample label with pre-printed Sample  ID
                                Clear tape strips
 ^                              Field Measurement Form
 p                              Pencils (for data forms) and Fine tipped indelible markers for labels
 CO
 t5
 §        5.3     Sampling Procedure
 ^        Table 5.2 presents step-by-step procedures for collecting microcystin samples at wadeable sites.

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             Wadeable
Table 5.2 Procedure: Microcystin Sample Collection (Wadeable Sites)
Microcystin Sample Collection
  1.   Fill out the pertinent information (Site ID and date) on the microcystin label and affix the label to
      the 500 mL Nalgene bottle. Completely cover the label with clear tape.
  2.   Collect the microcystin samples from the X-site in a flowing portion near the middle of the site.
  3.   Put on nitrile gloves. Make sure not to handle sunscreen or other chemical contaminants until
      after the sample is collected.
  4.   Rinse the 3-L Nalgene beaker three times with water, and discard the rinse downstream.
  5.   Fill the 3 liter beaker with water and slowly pour 30 - 50 ml into the 500 ml Nalgene bottle. Cap
      the bottle and rotate so that the water contacts all the surfaces. Discard the water downstream.
      Repeat this rinsing procedure  2 more times.
  6.   Fill the beaker with water and pour into the 500 ml Nalgene bottle. Leave one inch of head space
      in the bottle  to allow for expansion when frozen.
Place the 500 ml Nalgene bottle in a cooler (on ice or water) and shut the lid. If a cooler is not available,
place the cube container in an opaque garbage bag and immerse it in the stream.
Record the Sample ID on the Sample Collection Form along with the pertinent site information (site
name, ID, date, etc.).
Upon returning to your base site (hotel, lab, office, etc.), freeze the entire sample and keep frozen until
shipping.
                                                                                                        O
                                                                                                        C£
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 QC
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 QC
 LJJ
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 QC
 u
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          6      BENTHIC MACROINVERTEBRATES

          6.1    Summary of Method
          Collect benthic macroinvertebrate composite samples using a D-frame net with 500 u.m mesh
          openings. Take the samples from the sampling stations at the 11 transects equally distributed
          along the targeted reach. Multiple habitats will be encountered and sampled using this
          approach. They include bottom substrate as well as woody debris, macrophytes, and leaf packs.
          Composite all sample material and field-preserve with ~95% ethanol.

          6.2    Equipment and Supplies
          Table 6.1 shows the checklist of equipment and supplies required to complete the collection of
          benthic macroinvertebrates. This checklist is similar to the checklist presented in Appendix A,
          which is used at the base location to ensure that all of the required equipment is brought to the
          site. Record collection data on the back of Sample Collection  Form (Figure 6.1).
          Table 6.1 Equipment and Supplies: Benthic Macroinvertebrate Collection at Wadeable Sites
          For collecting     Modified kick net (D-frame with 500 urn
          samples          mesh) and 4-ft handle
                          Watch with timer or stopwatch
                          Buckets, plastic, 8 to 10 qt (optional)
                          Sieve bucket with 500 u.m mesh openings
                          (U.S. std No. 35)
                          Watchmakers' forceps
                          Wash bottle, 1-L capacity labeled "STREAM
                          WATER"
                          Funnel, with large bore spout


          For recording     Composite benthic sample labels with &
          measurements    without preprinted ID Sample ID numbers
                          Blank labels on waterproof paper for inside of
                          jars
Small spatula, spoon, or scoop to
transfer sample
Sample jars, 1-L HOPE plastic suitable
for use with ethanol
95% ethanol, in a proper container
Cooler (with absorbent material) for
transporting ethanol & samples
Plastic electrical tape
Scissors
Field Operations Manual or laminated
Quick Reference Guide

Soft (#2) lead pencils
Fine-tip indelible markers
Clear tape strips
Sample Collection Form
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                                                                        Field Operations Manual
                                                                                        Wadeable
    SUBSTRATE CHAN
                                   NRSA 2013/14 SAMPLE COLLECTION (Back)

                                                                     Date:        /
    BENTHIC MACROINVERTEBRATES {BERW) - WADEABLE
                                                                                             No Sample Collected
     Sample ID
                    Preserved  No of
         Number of jars   IETOH)  Transects  Comments
                                       O
     REACH-WIDE BENTHOS - WADEABLE
      TRANSECT
    IT otlur fixg wd
    explain in comment!
                  OG
                  O °T
                      ORl
                      O BA
                     Flag
                           O OT
                               O GL
                               QK,
                             Flag
                                       O GL
                                       O"'
                               O ** O OT O "* O °T
                                      Flag
                                                O GL
                                                    OG
                                                O R" O°
                                              Flag
                                                             OG
                                                        O f» O CT
                                                       Rag
                                                                 O GL
                                                                     OG
                                                               Flag
                                                                 O R* O °T O R*
                                                                        Flag
                                                                                      OG
                                                             O OT O "* O OT
                                                                                Flag
                                                                                           O GL
                                                                                          O RA O °
                                                                                         Flag
                                                                                                   OP
                                                                                                        OG
                                                                                                  Flag
                                                                                                   O R» O OT O R*
                                                                                                          Flag
    BENTHIC MACROINVERTEBRATES (BETB) - BOATABLE
                                                                             No Sample Collected Q
     Sample ID
                                     Preserved   Mo. of
                         Number of jars  (ETCH)  Transects Comments
    TRANSECT BENTHOS - BOATABLE

    Habitat: C = Coarse Subslrate / LWD L = Leaf Pack F = Organic Fine Muds / Sand M = Macrophyte beds OT = Other (Fbg wd ».pi»in n commwt s«c«on btiaw)
    Substrate:  F = Fine / Sand G = Gravel C = Coarse substrate OT = Other {Flag and explain n  comment section betow]
    Channel: P = Pool GL = Glide RI = Riffle RA= Rapid OT = Other (Flag and explain in comment section below)
      TRANSECT
    Location (UK}:
        Dominant
        Habitat:
        Secondary
         Habitat:
     Flag
                 OF  O"
                  Q or
Oc QL
OF O«
 O o7
                 OF  Oc
                 O G  O O
                 O p O"»
                 O RJ o GL
                  Q QT
                             B
                              Qs
                         Oc
                              O"
                          Q OT
                         O f  O M
                           O OT
         OF  Qc

         O G  O
         O f  O
         O Rl O
          Q QT
                            Flag
                 Qi
                 OF  O"
                  Q oi
OF
 O
OF Qc

O G O
O Rl O GL
 Q OT
                                    Flag
                              OB
        Oc  OL
        OF  O"
          Q OT
OF  O«
 O OT
OF Qc
O G O
OP O™>
O Rl O
 Q QT
                                             Flag
                                      O"
        Oc  OL
        OF  O"
          QOT
Oc OL
OF O«
 OOT
                                                  O G  O
OP OF
O "I O
 Q OT
                                                     Flag
                                                               O"
Oc C
OF C
 OOT
                 Oc
                 OF
                  Q or
Oc OL
OF O«
 O OT
Oc OL
OF QH
 OOT
        OF Qc

        O G O
OP Of
O Rl O G
 Q OT
                                                             Hag
                                                                        QB
                                             H
                                         OL.  OB
                OF  O"
                 Q OT
Oc OL
OF OM
 OOT
        OF

        O G
        OP Of
        ORI Ot
         Q OT
        OF Q

        O G O OT
OP Of
ORlOt
 Q OT
                                                                              Flag
                                                                                        I
                 Oc  OL
                 OF  O"
                  Q OT
Oc OL
OF O«
 O OT
        OF Qc

        O G O OT
                 OF O"W
                 O Rl O GL
                  Q OT
                                                                                      Flag
                                                                                             OL
                 Oc  OL
                 OF  O»
                  Q OT
                                  Oc  OL
                                  OF  OM
                                   OOT
        OF  Qc

        O G  O OT
                 OP OR*
                 O Rl O GL
                  Q QT
                                                                                               Flag
                                                                                                         K
                                                                                          OB
                  Oc  OL
                  OF  O»
                   Q c-
                           Oc  OL
                           OF  O"
                            OOT
                                           OF Qc

                                           O G O OT
                  OP OR»
                  O Rl O GL
                   Q QT
                                                                                              0629051193
           04/08/2013 2013 Sample Collection
Figure 6.1 Sample Collection Form (back)
                                                                                                               CO
                                                                                                               £
                                                                                                               o:
                                                                                                               LLJ

                                                                                                               Z
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                                                    Wadeable
 QC
 CD
 QC
 LJJ
 o
 QC
 u
 u
          6.3    Sampling Procedure
          Figure 6.2 summarizes how samples will be collected from wadeable sites. The transect sample
          design for collecting benthic macroinvertebrates is shown in Figure 6.3. Collect a sample from 1-
          m downstream of each of the 11 cross-section transects at the assigned sampling station. The
          process for selecting the sample stations is described in the Initial Site Procedures (Section 0). At
          transects assigned a "Center" sampling point where the stream width is between one and two
          net widths wide, pick either the "Left" or "Right" sampling point instead. If the stream is only
          one net wide at a transect, place the net across the entire stream width and consider the
          sampling point to be "Center". If a sampling point  is located in water that is too deep or unsafe
          to wade,  select an alternate sampling point on the transect at random.
          The procedure for collecting samples at each transect is described in Table 6.2. At each sampling
          point, determine if the habitat is a "riffle/run" or a "pool/glide" (any area where there is not
          sufficient current to extend the net is operationally defined as a pool/glide habitat). Record the
          dominant substrate type (fine/sand, gravel, coarse substrate (coarse gravel or larger) or other
          (e.g., bedrock, hardpan, wood, aquatic vegetation, etc.) and the habitat type (pool, glide, riffle,
          or rapid) for each sample collected on  the Sample  Collection Form as shown in Figure 6.1. As
          you proceed upstream from transect to transect, combine all samples into a bucket.
                                                  WADEABLE
                                         At Transect A, randomly locate 1st
                                         sampling point (left, center, or right
                                               facing downstream)
                                        Collect sample using riffle/run or pool/
                                         glide procedures from 25%, 50%, &
                                             75% width of the channel
                                            Transfer sample into bucket
                                         Mark bubble for substrate & channel
                                         habitat type on the Sample Collection
                                                     Form.
                                       Thoroughly rinse net and proceed to the
                                              next sampling location.
Composite the samples from all stations
 to create a single sample for the site.
          Figure 6.2 Benthic Macroinvertebrate Collection at Wadeable Sites
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           Wadeable
        K
                    Combine ALL kick net samples collected from ALL transects
                                             I
                             TRANSECT SAMPLES (1 per transect)
              Sampling point at each transect selected systematically after random start
                       Sampling points proceed in L, C, R pattern upstream
                                   Modified D-frame kick net
                              1 square foot quadrat sampled for 30 seconds
                                             I
                        COMPOSITE SAMPLES FROM ALL TRANSECTS
                               •  Seive bucket or other bucket(s)


1
SIEVE SAMPLE _
• 500 urn sieve bucket ^^H
• Remove and wash large objects ^^^1
1
COMPOSITE AND PRESERVE SAMPLE
• 1 liter bottle(s) (max of 4 bottles if possible)
• Fill no more than 50% with sample
• Preserve with ~95% ethanol for a final con-
centration of at least 70%
V

>


1 L

N
)

Figure 6.3 Transect Sample Design for Collecting Benthic Macroinvertebrates at Wadeable Sites
                                                                                             cc.
                                                                                             CO
                                                                                             CC
                                                                                             LJJ
                                                                                             o
                                                                                             cc.
                                                                                             u
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                                                                       Field Operations Manual
                                                                                    Wadeable
          Table 6.2 Procedure: Benthic Macroinvertebrates (Wadeable Sites)
 cc.
 CO
 CC
 LJJ
 o
 cc.
 u
 u
          Collecting Benthic Macroinvertebrate Sample
             1.  At 1 m downstream of each transect, beginning with Transect "A", randomly locate the first sampling
                station (Left, Center, or Right as you face downstream) as 25%, 50%, and 75% of the wetted width,
                respectively. If you cannot collect a sample at the designated point because of deep water or unsafe
                conditions, relocate to another random point on the same transect.
             2.  Determine if there is sufficient current in the area at the sampling station to fully extend the net. If
                so, classify the habitat as "riffle/run" and proceed to Step 3. If not, use the sampling procedure
                described for "pool/glide" habitats starting at Step 9.
              NOTE: If the net cannot be used, hand-pick a sample for 30 seconds from about 1ft2 of substrate at the
              sampling point. For vegetation choked sampling points, sweep the net through the vegetation  within a 1
              ft2 quadrat for 30 seconds. Place this hand-picked sample directly into the sample container. Assign a
              "U" flag (non-standard sample) to the sample and indicate which transect(s) required the modified
              collection procedure in the comments section. Go to Step 13.
           Riffle/Run Habitats:
            5.
3.   With the net opening facing upstream, quickly position the net securely on the stream bottom to
    eliminate gaps under the frame. Avoid large rocks that prevent the net from seating properly on the
    stream bottom.
  NOTE: If there is too little water to collect the sample with the D-net, randomly pick up 10 rocks from the
  riffle and pick and wash the organisms off them into a bucket which is half full of water.
4.   Holding the net in position on the substrate, visually define a quadrat that is one net width wide and
    long upstream of the net opening. The area within this quadrat is 1 ft2
    Check the quadrat for heavy organisms, such as mussels and  snails.  Remove these organisms by hand
    and place them into the net. Pick up loose rocks or other larger substrate particles in the quadrat.
    Use your hands or a scrub brush to dislodge organisms and wash them into the net. Scrub all rocks
    that are golf ball sized or larger and which are at least halfway into the quadrat. After scrubbing,
    place the substrate particles outside of the quadrat.
    Hold the D-net securely in position. Starting at the upstream  end of the quadrat, vigorously kick the
    remaining finer substrate within the quadrat for 30 seconds (use a stopwatch).
  NOTE: For samples located within dense beds of long, filamentous aquatic vegetation (e.g., algae or
  moss), kicking within the quadrat may not be sufficient to dislodge organisms in the vegetation. Usually
  these types of vegetation are lying flat against the substrate due to current. Use a knife or scissors to
  remove only the vegetation that lies within the quadrat (i.e., not entire strands that are rooted within
  the quadrat) and place it into the net.
7.   Pull the net up out of the water. Immerse the net in the stream several times to remove fine
    sediments and to concentrate organisms at the end of the net. Avoid having any water or material
    enter the mouth of the net during this operation.
8.   Go to Step 13.
            6.
           Pool/Glide Habitats:
             9.
    Visually define a quadrat that is one net width wide and long at the sampling point. The area within
                   ..2
    this quadrat is 1ft
10. Check the quadrat for heavy organisms, such as mussels and snails. Remove these organisms by hand
    and place them into the net. Pick up loose rocks or other larger substrate particles in the quadrat.
    Use your hands or a scrub brush to dislodge organisms and wash them into the net. Scrub all rocks
    that are golf ball sized or larger and which are at least halfway into the quadrat. After scrubbing,
    place the substrate particles outside of the quadrat.
11. Vigorously kick the remaining finer substrate within the quadrat with your feet while dragging the net
    repeatedly through the disturbed area just above the bottom. Keep moving the net all the time so
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      that the organisms trapped in the net will not escape. Continue kicking the substrate and moving the
      net for 30 seconds.
    NOTE: If there is too little water to use the kick net, stir up the substrate with your gloved hands and use
    a sieve with 500 urn mesh size to collect the organisms from the water in the same way the net is used
    in larger pools.
  12.  After 30 seconds, remove the net from the water with a quick upstream motion to wash the
      organisms to the bottom of the net.
All samples:
  13.  Invert the net into a sieve bucket and transfer the sample. Remove as much gravel as possible so that
      the organisms do not get damaged. Inspect the net for any residual organisms clinging to the net and
      deposit them into the bucket. Use forceps if necessary to remove organisms from the net. Carefully
      inspect any large objects (such as rocks, sticks, and leaves) in the bucket and wash any organisms
      found off of the objects and into the bucket before discarding the object. Remove as much detritus as
      possible without losing organisms.
    NOTE: »lt is recommended that crews carry a sample bottle containing a small amount ofethanol with
    them to enable them to immediately preserve larger predaceous invertebrates such as hellgrammites
    and water beetles. Doing so will help reduce the chance that other specimens will be consumed or
    damaged prior to the end of the field day.
  14.  Determine the predominant substrate size/type you sampled within the sampling quadrat. Mark the
      appropriate substrate type for the transect on the Sample Collection Form. The substrate types are:
      •   Fine/sand (F): not gritty (silt/clay/muck <0.06 mm diam.) to gritty, up to ladybug sized (2 mm)
      •   Gravel (G): fine to coarse gravel (ladybug to tennis ball sized; 2 mm to 64 mm)
      •   Coarse (C): Cobble to boulder (tennis ball to car sized; 64 mm to 4000 mm)
      •   Other (OT): bedrock (larger than car sized; > 4000 mm), hardpan (firm, consolidated fine
          substrate), wood of any size, aquatic vegetation,  etc.). Note type of "other" substrate in
          comments on field form.
    NOTE: If there are co-dominant substrate types, you may indicate more than one substrate type; note
    the co-dominants in the comments section of the form.
  15.  Identify the channel habitat type where the  sampling quadrat was located. Indicate the appropriate
      channel habitat type for the transect on the Sample Collection  Form. The channel habitat types are:
      •   Pool (P): Still water; low velocity; smooth, glassy  surface; usually deep compared to other parts
          of the channel
      •   Glide (GL): Water moving slowly, with smooth, unbroken surface; low turbulence
      •   Riffle (Rl): Water moving, with small ripples, waves, and eddies; waves not breaking, and surface
          tension is not broken; "babbling" or "gurgling" sound.
      •   Rapid (RA): Water movement is rapid and turbulent; surface with intermittent "white water"
          with breaking waves; continuous rushing sound.
      •   Other (OT): Note type of "other" channel habitat in comments on field form.
  16.  Thoroughly rinse the net before proceeding  to the next sampling station. Proceed upstream to the
      next transect (through Transect K, the upstream end of the reach) and repeat steps 1-15. Combine
      all kick net samples from riffle/run and pool/glide habitats into the bucket.
  17.  Record  the number of transects that were sampled throughout the reach.

6.4    Sample Processing in Field

Use a 500 urn mesh sieve bucket placed inside a larger bucket full of site water while sampling
to carry the composite sample as you travel around the site. Once the composite sample from
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          all stations is sieved and reduced in volume, store in a 1-liter jar and preserve with 95% ethanol.
          Do not fill jars more than 1/£ full of material. Multiple jars may be required if detritus is heavy
          (Table 6.3). Try to use no more than 4 jars per site. If more than one jar is used for a composite
          sample, use the "extra jar" label provided; record the SAME sample ID number on this "extra
          jar" label. DO NOT use two different sample numbers on two jars containing one single
          sample. Cover the labels with clear tape. The sample ID number is also recorded with a No. 2
          lead pencil on a waterproof label that is placed  inside each jar. Be sure the inside label and
          outside label describe the same sample.
          Record information for each composite sample on the Sample Collection Form as shown in
          Figure 6.1. Place the samples in a cooler or other secure container for transporting and/or
          shipping to the laboratory (see Appendix C). If the sample contains primarily organic material, or if
          adverse weather conditions exist (i.e. hot humid weather) process the sample at each station by placing it
          in a 1-L Nalgene jar with ethanol.
          Table 6.3 Procedure: Compositing Samples for Benthic Macroinvertebrates (Wadeable Sites)
                                   Compositing Benthic Macroinvertebrate Sample
            1.   Pour the entire contents of the bucket into a sieve bucket with 500 u.m mesh size. Remove any large
                objects and wash off any clinging organisms back into the sieve before discarding. Remove any large
                inorganic material, such as cobble or rocks.
            2.   Using a wash bottle filled with river water, rinse all the organisms from the bucket into the sieve.
                This is the composite sample for the reach.
            3.   Estimate the total volume of the sample in the sieve and determine how many 1-L jars will be
                required. Try to use no more than 4 jars per site.
            4.   Fill in a sample label with the Sample ID and date of collection. Attach the completed label to the jar
                and cover it with a strip of clear tape. Record the sample ID number for the composite sample on
                the Sample Collection Form. For each composite sample, make sure the number on the form
                matches the number on the label.
            5.   Wash the contents of the sieve to one side by gently agitating the sieve in the water. Wash the
                sample into a jar using as little water from the wash bottle as possible. Use a large bore funnel  if
                necessary. If the jar is too full pour off some water through the sieve until the jar is not more than K
                full, or use a second jar if necessary. Carefully examine the sieve for any remaining organisms and
                use watchmakers' forceps to place them into the sample jar.
                  •   If a second jar is needed, fill in a sample label that does not have a pre-printed ID number on it. Record
                      the ID number from the pre-printed label prepared in Step 4 in the "SAMPLE ID" field of the label. Attach
                      the label to the second jar and cover it with a strip of clear tape. Record the number of jars required for
                      the sample on the Sample  Collection Form. Make sure the number you record matches the actual
                      number of jars used. Write "Jar N ofX" on each sample label using a waterproof marker ("N" is the
                      individual jar number, and "X" is the total number of jars for the sample).
            6.   Place a waterproof label inside each jar with the following information written with a  number 2 lead
                pencil:
        Site ID
        Type of sampler and mesh size used
        Name of site
        Date of collection
Collectors initials
Number of stations sampled
Jar"N"of"X"
7.   Completely fill the jar with 95% ethanol (no headspace). It is very important that sufficient ethanol
    be used, or the organisms will not be properly preserved. Existing water in the jar should not dilute
    the concentration of ethanol below 70%.
    NOTE: Composite samples can be transported back to the vehicle before adding ethanol if necessary. In this
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                          Compositing Benthic Macroinvertebrate Sample
           case, fill the jar with stream water, which is then drained using the net (or sieve) across the opening to
           prevent loss of organisms, and replace with ethanol.
  8.   Replace the cap on each jar. Slowly tip the jar to a horizontal position, then gently rotate the jar to
      mix the preservative. Do not invert or shake the jar. After mixing, seal each jar with plastic tape.
  9.   Store labeled composite samples in a container with absorbent material that is suitable for use with
      70% ethanol until transport or shipment to the laboratory.
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          7      PERIPHYTON

          7.1    Summary of Method
          Collect periphyton from the 11 cross-section transects ("A" through "K") established within the
          sampling reach. Collect periphyton samples at the same transect location (L, C, or R) as the
          benthic macroinvertebrate samples (Section 6) directly after collecting the benthic
          macroinvertebrate sample. Prepare one composite "index" sample of periphyton for each site.
          At the completion of the day's sampling activities, but before leaving  the site, prepare three
          types of laboratory samples (an ID/enumeration sample to determine taxonomic composition
          and relative abundances, a chlorophyll sample, and a biomass sample (for ash-free dry mass
          [AFDM]) from the composite periphyton sample.

          7.2    Equipment and Supplies
          Table 7.1 is a checklist of equipment and supplies required to conduct periphyton sample
          collection and processing activities. This checklist is similar to the checklist presented in
          Appendix A, which is used  at the base location to ensure that all of the required equipment is
          brought to the site.
          Table 7.1 Equipment and Supplies: Periphyton (Wadeable Sites)

          For collecting samples          Large Funnel (15-20 cm diameter)
                                      12-cm2 area delimiter (3.8 cm diameter pipe, 3 cm tall)
                                      Stiff-bristle toothbrush with handle bent at 90° angle
                                      1-L wash bottle for stream water
                                      500-mL graduated plastic bottle for the composite sample
                                      60-mL plastic syringe with tip removed, and length of tubing
                                      Field Operations Manual and a laminated  Quick Reference Guide

          For recording measurements    Sample Collection Form
                                      Soft (#2) lead pencils for recording data on field forms
                                      Fine-tipped indelible markers for filling out sample labels
                                      Sample labels (4 per set) with the same Sample ID Number
                                      Clear tape strips for covering labels

          7.3    Sampling Procedure
          At each of the 11 transects, collect samples from the sampling station assigned during the layout
          of the  reach (L, C, or R). Collect the substrate selected for sampling from a depth no deeper than
          0.5 m. If a sample cannot be  collected because the location is too deep, skip the transect. The
          procedure for collecting samples and preparing a composite sample is presented in Table 7.2.
          Collect one sample from each of the transects and composite in one bottle to produce one
          composite sample for each site. Record the volume of the composite sample on the Sample
          Collection Form as shown in Figure 4.2.

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Table 7.2 Procedure: Collecting Composite Index Samples of Periphyton (Wadeable Sites)
Periphyton Composite Sample
 1.   Starting with Transect "A", collect a single sample from the assigned sampling station using the
     procedure below.
        If coarse substrate (cobbles, woody materials, etc.) are present that can be removed from the
        water:
        (a)   Collect a sample of substrate (rock or wood) that is small enough (< 15 cm diameter) and can
             be easily removed from the water. Place the substrate in a plastic funnel which drains into a
             500-mL plastic bottle with volume graduations marked on it.
        (b)   Use the area delimiter to define a 12-cm2area on the upper surface of the substrate. Dislodge
             attached periphyton from the substrate within the delimiter into the funnel by brushing with
             a stiff-bristled toothbrush for 30 seconds. Take care to ensure that the upper surface of the
             substrate is the surface that is being scrubbed, and that the entire surface within the
             delimiter is scrubbed.
        (c)   Fill a wash bottle with river water. Using water from this bottle, wash the dislodged
             periphyton from the funnel into the 500-mL bottle. Use an amount of water (~45 mL) that
             brings the composite volume up to the next graduation mark on the bottle.
        (d)   Put the bottle in a cooler on ice while you travel between transects and collect the
             subsequent samples. (The samples need to be kept cool and dark because a chlorophyll
             sample will be filtered from the composite.)
        If large coarse substrate is present that is too large to remove from the water (bedrock, large
        woody materials, boulders, etc.):
        (a)   Use the area delimiter to define a 12-cm2area on the upper surface of the substrate. Dislodge
             attached periphyton from the substrate within the delimiter using the tip of the syringe in a
             scraping motion.
        (b)   While dislodging periphyton with the syringe tip, simultaneously pull back on the syringe
             plunger to draw the dislodged periphyton into the syringe.
        (c)   Empty the syringe into the same 500-mL plastic bottle as above. If the volume  of the
             vacuumed sediment is not enough to raise the composite volume to the next graduation on
             the bottle (~45  mL), add additional stream water to the bottle to raise the level to the next
             graduation.
        (d)   Put the bottle in a cooler on ice while you travel between transects and collect the
             subsequent samples. (The samples need to be kept cool and dark because a chlorophyll
             sample will be filtered from the composite.)
        If no coarse sediment (cobbles or larger)  are present:
        (a)   Use the area delimiter to confine a 12-cm2 area of soft sediments.
        (b)   Vacuum the top 1 cm of sediments from within the delimited area into a de-tipped  60-mL
             syringe.
        (c)   Empty the syringe into the same 500-mL plastic bottle as above. If the volume  of the
             vacuumed sediment is not enough to raise the composite volume to the next graduation on
             the bottle (~45  mL), add additional stream water to the bottle to raise the level to the next
             graduation.
        (d)   Put the bottle in a cooler on ice while you travel between transects and collect the
             subsequent samples. (The samples need to be kept cool and dark because a chlorophyll
             sample will be filtered from the composite.)
 2.   Repeat Step 1 for transects "B" through "K". Place the sample collected at each sampling site into the
     single 500-mL bottle to produce the composite index sample.
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           3.  After samples have been collected from all 11 transects, thoroughly mix the 500-mL bottle regardless
              of substrate type. Record the total volume of the composite sample in the periphyton section of the
              Sample Collection Form.
           4.  If all 11 samples are not collected, record the number of transects collected and reason for any
              missed collection on the field forms.
          7.4    Sample Processing in the Field
          You will prepare three different types of laboratory samples from the composite index samples:
          an ID/enumeration sample (to determine taxonomic composition and relative abundances), a
          chlorophyll sample, and a biomass sample (for ash-free dry mass [AFDM]). All of the methods
          for processing the three samples are found in the Final Site Activities section of the manual. All
          the sample containers required for an  individual site should be sealed in plastic bags until use to
          avoid external sources of contamination (e.g., dust, dirt, or mud) that are present at site
          shorelines.
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8
      PHYSICAL HABITAT CHARACTERIZATION
Field measurements for physical habitat are made at two scales of resolution along the mid-
channel length of the reach the results are later aggregated and expressed for the entire reach.
The protocol defines the length of each sampling reach proportional to river wetted width and
then systematically places measurements to statistically represent the entire reach.
8.1     Equipment and Supplies
Table 8.1 lists the equipment and supplies required to conduct all the activities described for
characterizing physical  habitat. This checklist is similar to the checklist presented in Appendix A,
which is used at the base location to ensure that all of the required equipment is brought to the
river. Use this checklist to ensure that equipment and supplies are organized and available at
the river site in order to conduct the activities efficiently.
Table 8.1 Equipment and Supplies: Physical Habitat
                   Convex spherical canopy densiometer (Lemmon Model B), modified with taped "V"
For making
measurements
 For recording
 data
                  GPS
                  1 roll each colored surveyor's flagging tape (2 colors)
                  2 pair chest waders
                  1 or 2 fisherman's vest with lots of pockets and snap fittings.
                  Digital camera with extra memory card & battery
                  50 m or 100 m measuring tape with reel
                  Meter stick for bank angle measurements
                  Laser rangefinder (400 ft. distance range) and clear waterproof bag
                  Clinometer
                  Binoculars
                  Bearing compass (Backpacking type)
                  Current velocity meter, probe, and operating manual
                  Top-set wading rod for use with current velocity meter
                  Surveyor's telescoping leveling rod
                  Sounding rod or wading staff
                  Level tripod
                  CST Berger SAL 20 Automatic Level
                  Field Operations Manual and/or laminated Quick Reference Guide
                  2 covered clipboards (lightweight, with strap or lanyard)
                  Soft (#2) lead pencils
                  11 plus extras Channel/Riparian Transect forms
                  11 plus extras Thalweg Profile forms
                  1+ extras field Form: Stream Verification Form
                  1+ extras field Form: Field Measurement Form
                  1+ extras field Form: Sample Collection Form
                  1+ extras field Form: Channel Constraint Form
                  1+ extras field Form: Visual Assessment Form
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         8.2     Summary of Methods Approach
         Physical habitat in streams includes all those physical attributes that influence or provide
         sustenance to organisms within the stream. The physical habitat of a stream varies naturally,
         thus expectations differ even in the absence of anthropogenic disturbance. The procedures are
         employed on a reach length 40 times its baseflow wetted width. Measurement points are
         systematically placed to statistically represent the entire reach. Stream depth and wetted width
         are measured at very tightly spaced intervals, whereas channel cross-section profiles, substrate,
         bank characteristics and riparian vegetation structure are measured at larger intervals. Woody
         debris is tallied along the full length of the sampling reach, and discharge is measured at one
         location. The tightly spaced depth and width measures allow calculation of indices of channel
         structural  complexity, objective classification of channel units such as pools, and quantification
         of residual pool depth, pool volume, and total stream volume.

         8.3     Components of the Habitat Characterization
         There are  six components of the physical habitat characterization (Table 8.2). Measurements
         are recorded on 11 copies of a two-sided field form, and separate forms for recording slope and
         bearing measurements, assessing the degree of channel constraint, recording evidence of debris
         torrents or recent major flooding, and for stream discharge measurements.
         Table 8.2 Summary of Components of Physical Habitat Characterization at Wadeable Sites
Component Description
Thalweg Profile
Wetted Width /
Bar Width
Woody Debris
Tally
Channel and
Riparian
Characterization
Assessment of
Channel
Constraint, Debris
Torrents, and
Major Floods
Discharge
• Measure maximum depth, classify habitat and pool forming features, and
check presence of backwaters, side channels and loose, soft deposits of
sediment particles at 10-15 equally spaced intervals between each of 11
transects (100 or 150 individual measurements along entire reach).
• Measure wetted width and bar width (if present) and evaluate substrate
particle size classes at 11 cross-section transects and midway between them
(21 width measurements and substrate notations along entire reach).
• Between each of the channel cross-sections, tally large woody debris numbers
within and above the bankfull channel according to specified length and
diameter classes (10 separate tallies).
• At 11 transects placed at equal intervals along reach:
• Measure channel cross-section dimensions, bank height, bank undercut
distance, bank angle, slope and compass bearing (back sight), and riparian
canopy density (with densiometer).
• Visually Estimate3: substrate size class and embeddedness; areal cover class
and type (e.g., woody trees) of riparian vegetation in Canopy, Mid-Layer and
Ground Cover; areal cover class offish concealment features, aquatic macro-
phytes and filamentous algae.
• Observe & Record3: Presence and proximity of human disturbances.
• After completing thalweg and transect measurements and observations,
identify features causing channel constraint, estimate the percentage of the
channel margin that is constrained for the whole reach, and estimate the ratio
of bankfull/valley width. Check evidence of recent major floods and debris
torrent scour or deposition.
• Measure water depth and velocity at 0.6 depths at 15 to 20 equally spaced
intervals across one carefully chosen channel cross-section.
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                  •   In very small streams, measure discharge by timing the passage of a neutrally
                      buoyant object through a segment whose cross-sectional area has been
                      estimated or by timing the filling of a bucket.
  8 Substrate size class is estimated for a total of 105 particles taken at 5 equally spaced points along each
   of 21 cross-sections. Depth is measured and embeddedness estimated for the 55 particles located along
   the 11 regular transects A through K. Cross-sections are defined by laying the surveyor's rod or tape to
   span the wetted channel. Woody debris is tallied over the distance between each cross-section and the
   next cross-section upstream. Riparian vegetation and human disturbances are observed 5m upstream
   and 5m downstream from the cross-section transect. They extend shoreward 10m from left and right
   banks. Fish cover types, aquatic macrophytes, and algae are observed within the channel 5m upstream
   and 5m downstream from the cross-section stations. These boundaries for visual observations are
   estimated by eye.

8.4     Work Flow for the Physical Habitat Components
The six components (Table 8.2) of the habitat characterization are organized into four grouped
activities described below in the following sections.

8.4.1   Thalweg Profile and  Large Woody Debris Tally
Thalweg spacing  is calculated such that either 10  or 15 evenly spaced measurements are made
between each transect (see section 8.5). Two people proceed upstream from the downstream
end of the sampling reach making observations and measurements at the calculated increment
spacing. One person is in the channel making width and depth measurements and determining
whether soft/small sediment deposits are present under his/her wading  staff. The other person
records these measurements, classifies the channel habitat, records presence/absence of side
channels and off channel habitats (e.g., backwater pools, sloughs, alcoves), and tallies large
woody debris.  Each time this crew reaches a flag marking a new cross-section transect, they
start filling out a  new copy of the Thalweg Profile and Woody Debris Form. They interrupt the
thalweg profile and woody debris tallying activities to complete data collection at each
cross-section transect as they come to it. When the crew member in the water makes a width
measurement at channel locations midway between regular transects (i.e. at the fifth  or seventh
thalweg measurement in each sub-reach), she or he also locates and estimates the size class of
the substrate particles on the left channel margin (0%) and at positions 25%, 50%, 75%, and
100% of the distance across the wetted channel. Procedures for this substrate tally are the same
as for those at regular cross-sections, but data are recorded on the thalweg profile side of the
field form.

8.4.2   Channel/Riparian Cross-Sections                                                       ^
                                                                                                Q
At each of the  11 transects, one person proceeds with the channel cross-section dimension and           \-
substrate notations as described above; and also  makes measurements of bank angle  (using rod          Cd
and clinometer) and canopy cover (using densitometer). The second person records those                ^
measurements on the Channel/ Riparian Cross-section Form while making visual estimates of            <
riparian vegetation structure, instream fish cover, and human disturbance specified on that              <
form. Slope is measured by measuring the difference in elevation between each transect and            u
bearing is determined by backsighting to the previous transect. Supplementary points may need          
modified by delaying the measurements for slope and bearing and the woody debris tally until           a.

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         after reaching the upstream end of the reach. Backsighting and wood tallies can be done on the
         way back downstream (Note that in this case, the slope and bearing data form would have to be
         completed in reverse order).

         8.4.3   Channel Constraint and Torrent Evidence
         After completing observations and measurements along the thalweg and at all 11 transects, the
         field crew completes the overall reach assessments of channel constraint and evidence of debris
         torrents and major floods.

         8.4.4   Stream Discharge
         Discharge measurements can be performed during or at the end of the field sampling, but need
         to be made after collecting the water chemistry sample so as not to disturb the water and
         compromise the sample. The measurements are taken at a chosen optimal cross-section (but
         not necessarily at a sampling transect) near the X-site. However, do not use the electromagnetic
         current meter close to where electrofishing is taking place. Furthermore, if a lot of channel
         disruption is necessary and sediment must be stirred up, wait on this activity until all chemical
         and biological sampling has been completed.

         8.5     Habitat Sampling Locations within the Reach
         Measurements are made at two scales along the length of the reach; the results are later
         aggregated for the entire reach. Figure 8.1 illustrates the locations within the reach where data
         for the different components of the physical habitat characterization are collected. Most
         channel and riparian features are characterized on 11 cross-sections and pairs of riparian plots
         spaced at 4 channel width intervals (i.e., transect spacing = l/10th the total reach length). The
         thalweg profile measurements must be spaced evenly over the entire reach. In addition, they
         must be sufficiently close together that they do not miss deep areas and major habitat units.
         Follow these guidelines for choosing the increment between thalweg profile measurements:
         Channel Width < 2.5m:

             •    Minimum reach length of 150 m is used

             •    Thalweg increment = 1.0 m (resulting in 15 thalweg measurements per subreach);

             •    A total of 150 evenly spaced thalweg profile measurements will be made, 15 between
                 each channel cross-section
 •z.
 p           •    Mid-subreach measurements are made at the 7th thalweg location
 g       Channel Width 2.5 to 3.5m:
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 <           •    Thalweg increment = 1.5 m (resulting in 10 thalweg , measurements per subreach);
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 <
                 each channel cross-section
CO
            •   Mid-subreach measurements are made at the 5th thalweg location
                                                          th
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    •   Thalweg increment = 0.01 x reach length (resulting in 10 thalweg measurements per
       subreach).

    •   A total of 100 evenly spaced thalweg profile measurements will be made, 10 between
       each channel cross-section

    •   Mid-subreach measurements are made at the 5th thalweg location
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                                ••*•--*,,
                                      •*-C"""^"-
                                          ***%Channely
                                             Channel/Riparian
                                               Cross-section.
           Thalweg
            profile
           stations
                                                                   , f C )        Woody
                                                                   '"•—^         Debris
                                                                                 Tally
                                                                       . ( B )/(between
                                                                               transects)
Intermediate transects {width and
substrate measurements only
                                                     Downstream end
                                                     of sampling reach
                                                                                 PRK/DVP SMS
         Figure 8.1 Reach Layout for Physical Habitat Measurements (plan view)
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8.5.1  Thalweg Profile and Large Woody Debris Measurements

8.5.1.1 Thalweg Profile
Thalweg refers to the flow path of the deepest water in a stream channel. The thalweg profile is
a longitudinal survey of maximum flow path depth. Data from the thalweg profile allows
calculation of indices of residual pool volume, stream size, channel complexity, and the relative
proportions of habitat types such as riffles and pools.
The procedure for obtaining thalweg profile measurements is presented in Table 8.3. Record
data on the Thalweg Profile and Woody Debris Data Form as shown in Figure 8.2. Use the
surveyor's rod and a metric ruler or calibrated rod or pole to make the required depth and width
measurements at each station, and to measure off the distance between stations as you
proceed upstream. You may need to make minor adjustments to align each 10th measurement
to be one increment short of the next transect. In streams with average widths less than 2.5 m,
make thalweg measurements at 1 meter increments. Because the minimum reach length is set
at 150 meters, there will be 15 measurements on a field data form: Station 0 at the transect plus
14 additional stations between it and the next transect upstream. Use the five extra lines on the
thalweg profile portion of the data form (Figure 8.2) to record measurements 10-14.
Measure thalweg depths at all stations. Flag any missing measurements using a K code and
explain the reason in the comments section of the field data form. At points where a direct
depth measurement cannot be made, make your best estimate of the depth, record it on the
field form, and flag the value using a U code (nonstandard measurement), explaining that it is an
estimated value in the comments section of the field data form. Where the thalweg points are
too deep for wading, measure the depth by extending the surveyor's rod or weighted line at an
angle to reach the thalweg point. On the field form enter a U code in the thalweg depth column
(to indicate a nonstandard technique), and record the water level on the rod or line, and the
rod/line angle, as determined using the external scale on the clinometer (vertical = 90) in the
comment field (i.e. "depth of xx cm taken at an angle of xx degrees"). If a direct measurement of
the thalweg depth is not possible, make the best estimate you can of the depth, record it, and
use a U flag and a comment to note it is an estimated value.

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          Table 8.3 Procedure: Thalweg Profile

              1.  Determine the increment distance between measurement stations based on the wetted width used
                 to determine the length of the reach. Using a laser rangefinder or surveyor's rod:
                 •    For widths < 2.5 m, establish stations every 1 m (150 total).
                 •    For widths > 2.5 and <3.5 m, establish stations every 1.5 m (100 total).
                 •    For widths > 3.5 m, establish stations at increments equal to 0.01 times the reach length (100
                      total).
              2.  Complete the header information on the Thalweg Profile and Woody Debris Form, noting the
                 transect pair (downstream to upstream). Record the increment distance determined in Step 1 in the
                 INCREMENT field on the field data form used for subreach A-B. Also record the total reach length on
                 this form. It is not necessary to repeat these calculations on the remainder of the Thalweg Profile
                 and Woody Debris forms.
              3.  Begin at the downstream end (station 0) of the first transect (transect A).
              4.  Measure the wetted width at station 0, and at either station 5 (if the stream width defining the
                 reach length is > 2.5 m), or station 7 (if the stream width defining the reach length is < 2.5 m).
                 Wetted width is measured across and  over mid-channel bars and boulders. Record the width on the
                 field data form to the nearest 0.1 m. For streams with interrupted flow, where no water is in the
                 channel at the station or transect, record zeros for wetted width.
                 NOTE: If a mid-channel bar is present at a station where wetted width is measured, measure the
                 wetted width across and including the bar, but also measure the bar width and record it on the field
                 data form.
              5.  At station 5 or 7 (see above) classify the size of the bed surface particle at the tip of your depth
                 measuring rod at the left wetted margin and at positions 25%, 50%, 75%, and 100% of the distance
                 across the wetted width of the stream. This procedure is identical to the substrate size evaluation
                 procedure described for regular channel cross-sections (transects A - K), except that for these
                 midway supplemental cross-sections, substrate size is entered on the thalweg profile side of the
                 field form.
              6.  At each thalweg profile station, use  a calibrated pole or  rod to locate the deepest point within the
                 deepest flow path (the thalweg), which may not always  be found at mid-channel (and may not
                 always be the absolute deepest point in every channel cross-section). Measure the thalweg depth to
                 the nearest cm from the substrate surface to the water surface, and record it on the thalweg profile
                 form. Read the depth on the side of the rod to avoid inaccuracies due to the wave formed by the
                 rod in  moving water.
                 NOTE: For streams with interrupted flow if there is no  water at a transect, record zeros for depth.
                 NOTE: Obtain thalweg depths at all stations. If the thalweg is too deep to measure directly, stand in
 O               shallower water and extend the surveyor's rod or pole at an angle to reach the thalweg. Determine
 H               the angle by resting the clinometer on the upper surface of the rod and reading the angle on the
 ^               external scale of the clinometer. Record a Uflag in the thalweg depth column to indicate a non-
 LU               standard procedure was used. Record the water level on the rod and the rod angle in the comments
 C               section of the field data form. For deeper depths, use the same procedure with a taut string as  the
 ^               measuring device. Tie a weight to one  end of a length  of string or fishing line, and toss the weight
 in               into the deepest channel location. Draw the string up tight and measure the length of the line that is
 i_               under  water. Measure the string angle with the clinometer exactly as done for the surveyor's rod. If a
 H               direct measurement cannot be obtained, make the best  estimate you can of the thalweg depth, and
 ^               use a Uflag to identify it as an estimated measurement.
 _,            7.  At the  point where the thalweg depth is determined, observe if unconsolidated, loose (soft)
 |5               deposits of small diameter (<16mm) sediments are present directly beneath your ruler, rod, or pole.
 S£               Soft/small sediments are defined here as fine gravel, sand, silt, clay or muck readily apparent by
 J               "feeling" the bottom with the rod. Record presence or absence in the SOFT/SMALL SEDIMENT field on


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       the field data form. Note: A thin coating of fine sediment or silty algae coating the surface of cobbles
       should not be considered soft/small sediment. However, fine sediment coatings should be identified
       in the comments section of the field form when determining substrate size and type.
   8.   Determine the channel unit code for the station. Record this on the field data form using the
       standard codes provided. For dry and intermittent streams, where no water is in the channel, record
       habitat type as dry channel (DR).
   9.   If the station cross-section intersects a mid-channel bar, indicate the  presence of the bar by filling in
       the bubble in the BAR WIDTH field on the field data form. However, a  measurement of the bar width
       is only taken if the bar intersects a transect at either the endpoint or  midpoint of a subreach.
   10. Record the presence or absence of a side channel at the station's cross-section in the SIDE CHANNEL
       field on the field data form.
   11. Record the presence or absence of quiescent off channel aquatic habitats, including sloughs, alcoves
       and backwater pools in the BACKWATER column of the field form.
   12. Proceed upstream to the next station, and repeat Steps 2 through 11.
   13. Repeat Steps 2 through 12 until you reach the next transect. At this point complete Channel/
       Riparian measurements at the new transect (Section 8.4.2). Then prepare a new Thalweg Profile and
       Woody Debris Form and repeat Steps 2 through 12 for each of the reach segments, until you reach
       the upstream end of the sampling  reach (transect K). At transect K, you will have completed 10
       copies of the Thalweg Profile and Woody Debris Form, one for each segment (A to 6, 6 to C, etc.).
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At every thalweg station, determine by sight or feel whether deposits of soft/small sediments
are present on the channel bottom. These particles are defined as substrate equal to or smaller
than fine gravel (< 16 mm diameter). These soft/small sediments are different from Fines
described when determining the substrate particle sizes at the cross-section transects (Section
8.7). If the channel bottom is not visible, determine if soft/small sediment deposits are readily
obvious by feeling the bottom with your boot, the surveyor's rod, or a calibrated rod or pole.
Measure wetted width at each transect (station 0), and midway between transects (station 5 for
larger streams having 100 measurement points, or station 7for smaller streams having 150
measurement points). The wetted width boundary is the point at which substrate particles are
no longer surrounded by free water. Estimate substrate size for five locations evenly spaced
across each midway cross-section using procedures described for substrate at regular cross-
sections (Figure 8.1), but at the supplemental cross-sections, only the size class (not distance
and depth) data are recorded.
While recording the width and depth measurements and the presence of soft/small sediments,
the second person evaluates and records the habitat class (Table 8.4) applicable to each of the
100 (or  150) measurement points along the length of the reach. Make channel unit scale habitat
classifications at the thalweg of the cross-section. The habitat unit itself must meet a minimum
size criteria in addition to the qualitative criteria listed in Table 8.4. Before being considered
large enough to be identified as a channel unit scale habitat feature, the unit should be at least
as long as the channel is wide. For instance, if there is a small deep (pool like) area at the
thalweg within a large riffle area, do not record it as a pool  unless it occupies an area about as
wide or long as the channel is wide. If a backwater pool dominates the channel, record PB as
the dominant habitat unit class. If the backwater is a pool that does not dominate the main
channel, or if it is an off channel alcove or slough (large enough to offer refuge to small fishes),
fill in the Y bubble to indicate presence of a backwater in the BACKWATER column of the field
form, but classify the  main channel habitat unit type according to characteristics of the main
channel. Sloughs are backwater areas having marsh like characteristics such as vegetation, and
alcoves  (or side pools) are deeper areas off the main channel that are typically wide and shallow
(Helm 1985, Bain and Stevenson 1999). When trying to identify the pool forming element for a
particular pool, remember that most pools are formed at high flows, so you  may need to look
for elements that are dry at baseflow, but still within the bankfull channel (e.g., boulders or
large woody debris).
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          Table 8.4 Channel Unit Categories
          Channel Unit Habitat Classes
          Class (Code)
                                 Description
          Pools: Still water, low velocity, a smooth, glassy surface, usually deep compared to other parts of the
          channel
          Plunge Pool (PP)
                                 Pool at base of plunging cascade or falls.
          Trench Pool (PT)
                                 Pool like trench in the center of the stream
          Lateral Scour Pool (PL)     Pool scoured along a bank.
          Backwater Pool (PB)      Pool separated from main flow off the side of the channel (large enough to
                                  offer refuge to small fishes). Includes sloughs (backwater with marsh
                                  characteristics such as vegetation), and alcoves (a deeper area off a wide
                                  and shallow main channel)
          Impoundment Pool (PD)   Pool formed by impoundment above dam or constriction.
          Pool (P)
                                 Pool (unspecified type).
          Glide (GL)
                                 Water moving slowly, with a smooth, unbroken surface. Low turbulence.
          Riffle (Rl)                Water moving, with small ripples, waves and eddies waves not breaking,
                                  surface tension not broken. Sound: babbling, gurgling.
          Rapid (RA)               Water movement rapid and turbulent, surface with intermittent Whitewater
                                  with breaking waves. Sound: continuous rushing, but not as loud as cascade.
          Cascade (CA)             Water movement rapid and very turbulent over steep channel bottom.
                                  Much of the water surface is broken in short, irregular plunges, mostly
                                  Whitewater. Sound: roaring.
          Falls (FA)                Free falling water over a vertical or near vertical drop into plunge, water
                                  turbulent and white over high falls. Sound: from splash to roar.
          Dry Channel (DR)
                                 No water in the channel, or flow is submerged under the substrate
                                 (hyporheicflow).
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          8.5.1.2 Large Woody Debris Tally
          Large Woody Debris (LWD) is defined here as woody material with a small end diameter of at
          least 10 cm (4 in.) and a length of at least 1.5 m (5 ft.). The procedure for tallying LWD is
          presented in Table 8.5. The tally includes all pieces of LWD that are at least partially in the
          baseflow channel (Zone 1), in the bankfull channel (Zone 2, flood channel up to bankfull stage),
          or spanning above the bankfull channel (Zone 3), as shown in Figure 8.3. LWD in zones 1 and 2
          will be tallied together (considered  all or in-part within the bankfull channel). The bankfull
          channel is defined as the channel that is filled by moderate sized flood events that typically
          recur every one to two years. LWD  in or above the bankfull channel is tallied over the entire
          length of the reach, including the area between the channel cross-section transects. Pieces of
          LWD that are not at least partially within Zones 1, 2, or 3 are not tallied.
          1 Note that in order for a channel habitat unit to be distinguished, it must be at least as wide or long as
          the channel is wide (except for off channel backwater pools, which are noted as present regardless of
          size .
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Table 8.5 Procedure: Tallying Large Woody Debris
 Large Woody Debris Tally Form
 Note: Tally pieces of large woody debris (LWD) within each segment of stream while the thalweg profile
 is being determined. Include all pieces in the tally whose large end is found within the subreach.
 1.  Scan the stream segment between the two cross-section transects where thalweg profile
     measurements are being made.
 2.  Tally all LWD pieces within the segment that are at least partially within the bankfull channel.
     Determine if a piece is LWD (small end diameter >10 cm [4 in.], and length  >1.5 m [5ft.])
 3.  For each piece of LWD, determine the class based on the diameter of the large end (0.1 m to < 0.3 m,
     0.3 m to <0.6 m, 0.6 m to <0.8 m, or >0.8 m), and the class based on the length of the piece (1.5m to
     <5.0m, 5m to <15m, or >15m).
     If the piece is not cylindrical, visually estimate what the diameter would be for a piece of wood with
       circular cross-section that would have the same volume.
     When estimating length, include only the part of the LWD piece that has a diameter >10 cm (4 in)
 4.  Place a tally mark in the appropriate diameter x length class tally box in the PIECES ALL/PART IN BANKFULL
     CHANNEL section of the Thalweg Profile  and Woody Debris Form.
 5.  Tally all LWD pieces within the segment that are not actually within the bankfull channel, but are at
     least partially spanning (bridging) the bankfull channel. For each piece, determine the class based on
     the diameter of the large end (0.1 m to < 0.3 m, 0.3 m to <0.6 m, 0.6 m to <0.8 m, or >0.8 m), and the
     class based on the length of the piece (1.5 m to <5.0 m, 5 m to <15 m, or >15  m).
 6.  Place a tally mark for each piece in the appropriate diameter x length class tally box in the PIECES
     BRIDGE ABOVE BANKFULL CHANNEL section of the Thalweg Profile and Woody Debris Form.
 7.  After all pieces within the segment have been tallied, write the total number  of pieces for each
     diameter x length class in the small box at the lower right-hand corner of each tally box.
 8.  Repeat Steps 1 through 7 for the next subreach, using a new Thalweg Profile and Woody Debris Form.
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           ZONE
                                     BANKFULL CHANNEL WIDTH
                                       ZONE 3
                                                    WATER SURFACE AT
                                                    BANKFULLFLOW
                                                           T
ZONE 4
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Figure 8.3 Large Woody Debris Influence Zones (modified from Robison and Beschta, 1990).

8.6    Channel and Riparian Measurements at Cross-Section Transects

8.6.1  Slope and Bearing
Measure slope and bearing by sighting between transects (e.g., transect 6 and A, C and 6, etc.).
To measure the slope between adjacent transects, follow the procedure presented in Table 8.6.
Measure bearing following the procedure presented in Table 8.8. Record slope and bearing data
on the Slope and Bearing Form as shown in Figure 8.4.
Slope is typically measured by two people, one holding a surveyor's rod and the second sighting
through the surveyor's level. Be sure that the person is holding the marked pole or rod at the
surface of the water. The intent is to get a measure of the water surface slope, which may not
necessarily be the same as the bottom slope. The surveyor's level is leveled according to the
manufacturer's recommendations, which is generally to  adjust the three leveling feet until the
bubble is centered. Level  is checked in all planes to be measured. If the level does not "self
level" in all measured planes the user should check the instruction manual for suggested
options. Elevation readings are made at each transect and the difference between each
elevation reading is recorded as the change in elevation. NOTE: Multiple transect elevations
can often be made for each setup of the level, but every time the tripod and level are moved a
second measurement of the last transect elevation from the last setup is required. You cannot
use elevations from previous setups because the relative height of the  level has changed.
To calculate sinuosity from bearing measurements, it does not matter whether or not you adjust
your compass bearings for magnetic declination, but it is important that  you are consistent in
the use of magnetic or true bearings throughout all the measurements you make on a given
reach. Note in the comments section of the Slope and Bearing Form which type of bearings you
are taking, so the measurements can be used to describe reach aspect. Also, guard against
recording reciprocal bearings (erroneous bearings 180 degrees from what they should be). The
best way to do this is to know where the primary (cardinal) directions are in the field: (north [0
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degrees], east [90 degrees], south [180 degrees], and west [270 degrees]), and insure that your
bearings "make sense."
As stated earlier, it may be necessary to set up intermediate (supplemental) bearing points
between a pair of cross-section transects if you do not have direct line-of-sight along (and
within) the channel between stations Figure 8.5. This can happen if brush is too heavy, or if
there are sharp slope breaks or tight meander bends. If you would have to sight across land to
measure bearing between two transects, then you need to make one or more supplemental
measurements (i.e., do not "short-circuit" a meander bend). Mark these supplemental locations
with a different color of plastic flagging than used for the cross-section transects to avoid
confusion. Record these supplemental bearing measurements, along with the proportion of the
stream segment between transects included in each supplemental measurement, in the
appropriate sections of the Slope and Bearing Form (Figure 8.4). Note that the main bearing
observations are always downstream of supplemental observations (i.e., to the downstream
transect).  Similarly, first supplemental observations are always downstream of second
supplemental observations.
Because measurements of slope are a calculation  of the elevation difference between transects,
you may sight over land for the purposes of slope only (Figure 8.5). You may need to use
supplemental points in your measurement of slope if visibility is severely limited, but
supplemental points are not required as they  are for measurements of bearing. As a result,
there may be times when you record bearing  data for a supplemental point (or two) on the
Slope and Bearing Form; but only record the total elevation change in the MAIN column.
Table 8.6 Procedure: Obtaining Slope Data
 Slope Method with Surveyors Level
 Instrument Setup:
     1.   Extend the tripod legs to approximately eye level and set the legs firmly into the ground; adjust
         the legs so that they form a regular triangle and are firmly set with no wobble. Adjust the legs so
         that the base plate is approximately level.
     2.   Hold the instrument on the tripod and start the centering screw. Ensure the adjustable feet are
         roughly evenly adjusted. While the centering screw is still loose slide the instrument on the base
         plate until the bubble is approximately centered in the circular level. Tighten the centering screw.
     3.   Adjust the leveling foot screws until the bubble is exactly level in the center circle.
     4.   Self-Leveling instruments can now be swiveled gently on the base plate and should maintain level
         as long as the tripod remains steady. Check to ensure the bubble indicates the instrument is level
         across all planes at which measurements are to be made.
     5.   Adjust focus, brightness and parallax according to manufactures specifications. The instrument is
         ready to make measurements.
 Taking Measurements:
 1.  Determine a location at transect K to hold a surveyor's rod that will be visible from a point between
     transect J and transect K:
     a.   Set up the instrument at a  point approximately halfway between transects J and K and where a
         clear line of sight is possible. If visibility allows, the instrument may be placed farther upstream, as
         long as transect K is still visible.
     b.   Position the staff at transect K, holding the bottom of the staff at the water level and the staff as
         vertical as possible and the numbers facing the instrument.
     c.   Site the staff through the instrument and record the reading to the nearest centimeter in a field
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                   notebook or other workspace (this value is not entered on the field form).
               d.  Move the staff to transect J and gently swivel the instrument to face the next reading. Hold the
                   staff as before, vertically, with the bottom at the water level and the numbers facing the
                   instrument.
               e.  Site the staff and record the reading to the nearest centimeter (again in a field notebook or
                   elsewhere).
               f.   Subtract the elevation reading at transect J from the reading at transect K.
               g.  The difference in the readings is the elevation  difference that  is recorded on the Slope and
                   Bearing Form.
               h.  Repeat measurements between each transect.
               Note: In small streams with a clear line of site it may be possible to set the instrument up once and
               make readings to several transects from a single set up. Simply record the readings for each transect
               and do not skip transects.
               • If you are back sighting to a supplemental point, you may record the elevation difference in the
                   appropriate SUPPLEMENTAL section  of the Slope and Bearing Form.
            2.  Proceed to the next cross-section transect (or supplementary point), and repeat Steps a - h above.
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           a Method codes are: C/.=clinometer, 77?=transit, H/.=hand level, l/l/T=Water tube, M=laser level,
            Omf/?=method not listed (describe in comments section of form).
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          Because of ease of use, portability, and cost, hand-held clinometers were previously used to
          determine slope. In this instance, the field crews will have access to more sophisticated
          instrumentation (e.g., surveyor's level), and have field personnel who are experienced in the use
          of these instruments. The Slope and Bearing Form (Figure 8.4) is designed to allow for different
          methods and/or different units of measuring slope.  Mark the appropriate method bubble
          (instead of Cl; method codes are identified in Table 8.6) and mark the % BUBBLE (instead of the
          CM bubble) if the method or instrument measures the percent slope rather than the change in
          elevation  (Table 8.7).
          Table 8.7 Modified Procedure: Obtaining Slope Data (without Surveyor's Level)
          Modified Slope Method - Only Use if Surveyor Level is NOT Possible
Use this procedure if you are starting at the upstream transect (K), after completing the thalweg profile
and other cross-section measurements at transects A through K. It should only be used if you cannot use
the surveyor's level.
    1.   Stand in the channel at the upstream cross-section transect. Determine if you can see the next
        cross-section  transect downstream.. If not, you may have to take supplementary slope
        measurements. While sighting over land is not prohibited during any slope measurement, the
        clinometer method is more difficult to use over larger distances, so supplementary
        measurements are more likely to be needed.
    2.   Mark a  surveyor's rod and a calibrated rod (or meter ruler) at the same height. If a shorter pole
        or ruler is used, measure the height from the ground to the opening of the clinometer when it is
        resting on top.
    3.   Have one person take the marked surveyor's rod to the downstream transect. Hold the rod
        vertical with the bottom at the same level as the water surface. If no suitable location is available
        at the stream margin, position the rod in the water and note the depth.
          If you have determined in Step 1 that supplemental measurements are required for this
          segment, walk downstream to the furthest point where you can  stand in the center of the
          channel and still see the center of the channel at the upstream cross-section transect.. Mark
          this location with a different color flagging than that marking the cross-section transects.
    4.   Place the base of the calibrated  rod at the level as the surveyor's rod (either at the water surface
        or at the same depth in the water).
    5.   Place the clinometer on the calibrated rod at the height determined in Step 2. With the
        clinometer, sight downstream to the flagged height on the surveyor's rod at the downstream
        transect (or at the supplementary point).
          If you are sighting to the next downstream transect, read and record the percent slope in the
          MAIN section on the Slope and Bearing Form for the downstream transect (e.g., J < K), which is
          at the bottom of the form (i.e., you are completing the form in reverse order). Record the
          PROPORTION as 100%.
          If you are backsighting from a supplemental point, record the slope (%) and proportion (%) of
          the stream  segment that is included in the measurement in the appropriate SUPPLEMENTAL
          section of the Slope and Bearing Form. The last sighting to a downstream transect (from either
          the upstream transect or the nearest upstream supplemental point) is always recorded as the
          MAIN reading.
    6.   Stand in the middle of the channel at upstream transect (or at a supplemental point), and sight
        with your compass to the middle of the channel at the downstream transect (or at a supplemen-
        tal point). Record the bearing (degrees) in the same section of the Slope and Bearing form
        (Supplemental or Main) as you recorded the slope in Step 6.
    7.   Proceed to the next  cross-section transect (or to a supplementary point), and repeat Steps 3
        through 7 above.
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                   Field Operations Manual
                                 Wadeable
        Backsight with
        compass and
        record
        main slope
        and bearing
        measurements
        and % of reach
Supplemental slope  Backsight with
and bearing point    compass and record
                  supplemental slope
                  and bearing
                  Measurements and
                  % of reach
Backsight
with compass
and record
main slope
and bearing
measurements
and % of reach
Figure 8.5 Measurements of Bearing Between Transects
                                                Short pole with
                                                clinometer at
                                                  height h
        Surveyor rod
       with flagging at
                                         ,«n <»*'
                                                     Upstream
                                                     Transect
                   Both poles must be at water's
                   surface or at same depth
  Downstream
    Transect
Figure 8.6 Channel Slope Measurement using a Clinometer
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          Table 8.8 Procedure: Obtaining Bearing Data
          Obtaining Bearing Data
          Use this procedure to backsight from one transect to another with the use of a sighting compass.
          Readings (in degrees) are taken from the center one of transect to the center of the next transect
          downstream.
          It does not matter whether you use true or magnetic bearings as long as you are consistent for each
          reading.
          Take care not to accidentally take a reciprocal reading (one that is 180 degrees off).
            1.   Stand in the center of the channel at the upstream cross-section transect. Determine if you can see
                the next cross-section transect downstream without sighting across land (i.e., do not "short-circuit"
                a meander bend). If not, you will have to take supplementary bearing measurements
            2.   Hold the compass in  line with your body and sight down the "lubber line" or through the sight
                window to the center of the next transect downstream (or supplemental point). It will help to have
                another person at the center of the transect to which you are sighting. Remember that your line of
                sight cannot "cross land."
            3.   While pointing the compass toward the middle of the transect or supplemental point, read the
                bearing (in degrees)
                •    For many navigational compasses, you will rotate the bezel until the index marks are centered
                    over magnetic north needle. The base of the lubber line or index mark on the sight now points
                    to the compass heading on the bezel.
                •    For some direct  reading compasses, the bearing will be displayed already.
            4.   Record the reading on the Slope and Bearing Form.
            5.   If you are sighting to the next downstream transect, read and record the bearing in the MAIN
                section on the Slope  and Bearing Form. Record the PROPORTION as 100%.
            6.   If you are backsighting from a supplemental point, record the bearing and proportion (%) of the
                stream segment that is included in the measurement in the appropriate SUPPLEMENTAL section of
                the Slope and Bearing Form. The last sighting to a downstream transect (from either the upstream
                transect or the nearest upstream supplemental point) is always recorded as the MAIN reading.
            7.   Proceed to the next transect (or supplemental point), and repeat steps 1 through 6 above.

          8.7    Substrate Size and Channel Dimensions

          Substrate size and embeddedness are evaluated at 5 points at each of the 11 transects (refer to
          Figure 8.7). Substrate size is also evaluated at 10 additional cross-sections located  midway
          between each of the 11 regular transects (A-K). In the process of measuring substrate particle
          sizes at each channel cross-section, the wetted width  of the channel and the water depth at
          each substrate sample point are measured (at the 10 midway cross-sections, only substrate size
          and wetted width are recorded). If the wetted channel is split by a  mid-channel bar (Section
          8.4.1), the five substrate  points are centered between the wetted width boundaries regardless
          of the mid-channel bar in between. Consequently, substrate particles selected in some
          cross-sections may be "high and dry". For cross-sections that are entirely dry, make
          measurements across the unvegetated portion of the channel.
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                                             Field Operations Manual
                                                         Wadeable
          Left
          Bank
 25%
Wetted
Width
  r
 50%
Wetted
Width
                                                                          Right
                                                                          Bank
                                                                           PRK/DVP sroe
Figure 8.7 Substrate Sampling Cross-Section

The substrate sampling points along the cross-section are located at 0, 25, 50, 75, and 100
percent of the measured wetted width, with the first and last points located at the water's edge
just within the left and right banks. The procedure for obtaining substrate measurements is
described in
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          National Rivers and Streams Assessment 2013/14                           Field Operations Manual
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          Table 8.9 (including all particle size classifications). Record these measurements on the
          Channel/Riparian Cross-section side of the field form, as shown in Figure 8.8.
          For the supplemental cross-sections midway between regular transects, record substrate size
          and wetted width data on the thalweg profile side of the field form. To minimize bias in
          selecting a substrate particle for size classification, it is important to concentrate on correct
          placement of the measuring stick along the cross-section, and to select the particle right at the
          bottom of the stick (not, for example, a more noticeable large particle that is just to the side of
          the stick). Classify the particle into one of the size classes listed on the field data form (Figure
          8.8) based on the middle dimension of its length, width, and depth. This median dimension
          determines the sieve size through which the particle can pass. When you record the size class as
          Other, assign an Fn flag on the field data form and describe the substrate type in the comments
          section of the field form, as shown in Figure 8.8.
          At substrate sampling locations on the 11 regular transects (A-K), examine particles larger than
          sand  for surface stains, markings, and algal coatings to estimate embeddedness of all particles in
          the 10 cm diameter circle around the substrate sampling point.  Embeddedness is the fraction of
          a particle's volume that is surrounded by (embedded in) sand or finer sediments on the stream
          bottom. By definition, record  the embeddedness of sand and fines (silt, clay, and muck) as 100
          percent, and  record the embeddedness of hardpan and bedrock as 0 percent.

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                                                                   Field Operations Manual
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Table 8.9 Procedure: Substrate Measurement
   1.
   2.
   3.
   4.
Fill in the header information on page 1 of a Channel/Riparian Cross-section Form. Indicate the
cross-section transect. At the transect, extend the surveyor's rod or metric tape across the channel
perpendicular to the flow, with the "zero" end at the left bank (facing downstream).
NOTE: If a side channel is present, and contains 16- 49% of the total flow, establish a secondary
cross-section transect. Use a separate field data form to record data for the side channel,
designating it as a secondary transect by marking both the EXTRA SIDE CHANNEL bubble and the
associated primary transect letter (e.g., A, B, etc.). Collect all channel and riparian cross-section
measurements from the side channel.
Divide the wetted channel width channel by 4 to  locate substrate measurement points on the cross-
section. In the DisrLB fields of the form, record the distances corresponding to 0% (LEFT), 25% (LCTR),
50% (CTR), 75% (RCTR), and 100% (RIGHT) of the measured wetted width. Record these distances at
Transects A-K, but just the wetted width at midway cross-sections.
Place your sharp ended meter stick or calibrated  pole at the LEFT location (0 m). Measure the depth
and record it on the field data form. (Cross-section depths are measured only at regular transects A-
K, not at the 10 midway cross-sections).
•   Depth entries at the left and right banks may be 0  (zero) if the  banks are gradual.
•   If the bank is nearly vertical, let the base of the measuring stick fall to the bottom (i.e., the
    depth at the bank will be > 0 cm), rather than holding it suspended at the water surface.
Pick up the substrate particle that is at the base of the  meter stick (unless it is  bedrock or boulder),
and visually estimate its particle size, according to the following table. Classify the particle according
to its median diameter (the middle dimension of  its length, width, and depth). Record the size class
code on the field data form. (Cross-section side of form for transects A-K; special entry boxes on
Thalweg Profile side of form for midway cross-sections.)
Code
RS
RR
HP
LB
SB
CB
GC
GF
SA
FN
WD
RC
OT
Size Class
Bedrock (Smooth)
Bedrock (Rough)
Hardpan
Boulders (large)
Boulders (small)
Cobbles
Gravel (Coarse)
Gravel (Fine)
Sand
Fines
Wood
Concrete
Other
Size Range (mm)
>4000
>4000
>4000
>1000 to 4000
>250 to 1000
>64 to 250
>16 to 64
> 2 to 16
>0.06 to 2
<0.06
Regardless of Size
Regardless of size
Regardless of Size
Description
Smooth surface rock bigger than a car
Rough surface rock bigger than a car
Firm, consolidated fine substrate
Yard/meter stick to car size
Basketball to yard/meter stick size
Tennis ball to basketball size
Marble to tennis ball size
Ladybug to marble size
Smaller than ladybug size; gritty between
fingers
Silt Clay Muck (not gritty between fingers)
Wood & other organic particles
Record size class in comment field
Metal, tires, car bodies etc. (describe in
comments)
   5.
Evaluate substrate embeddedness as follows at each of the five points. For particles larger than
sand, examine the surface for stains, markings, and algae. Estimate the average % embeddedness of
particles in the 10 cm circle around the measuring rod. Record this value on the field data form. For
sand and smaller particles, you will not be able to pick up an individual particle, but a "pinch" of fine
particles between your fingers. Determine and record the dominant size of particles in the "pinch."
By definition, sand and fines are  embedded 100%; bedrock and hardpan are embedded 0%.
                                                                                                         73

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          National Rivers and Streams Assessment 2013/14                           Field Operations Manual
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            6.  Move to the next location on the transect, and repeat Steps 3 - 5 at each location. Repeat Steps 1 - 5
                at each transect, including any additional side channel transects established if side channels are
                present.

          8.7.1   Bank Characteristics
          The  procedure for obtaining bank and channel dimension measurements is presented in Figure
          8.10. Data are recorded in the BANK MEASUREMENTS section of the Channel/Riparian Cross-section
          Form as shown in Figure 8.8. Bank angle and bank undercut distances are determined on the
          left and right banks at each cross-section transect. Figure 8.9 illustrates how bank angle is
          determined for several different situations. The scale at which bank angle is characterized is
          approximately 0.5 m. A short (approx. 1 m long) pole is used to determine bank angle. The angle
          is determined based on the pole resting on the ground for about 0.5 m. Other features include
          the wetted width of the channel (as determined in Section 8.4.1), the width of exposed mid-
          channel bars of gravel or sand, estimated incision height, and the estimated height and width of
          the channel at bankfull stage as described in Figure 8.10. Bankfull height and incised height are
          both measured relative to the present water surface (i.e. the level of the wetted edge of the
          stream). This is done by placing the base of the small measuring rod at the bankfull elevation
          and  sighting back to the survey rod placed at the water's edge using the clinometer as a level
          (i.e., positioned so the slope reading is 0%.). The height of the clinometer above the base of the
          smaller rod  is subtracted from the elevation sighted on the surveyor's rod. Bankfull flows are
          large enough to erode the stream bottom and banks, but frequent enough  (every 1 to 2 years)
          to not allow substantial growth of upland terrestrial vegetation. Consequently, in many regions,
          it is these flows that have determined the width and depth of the channel.

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National Rivers and Streams Assessment 2013/14
Draft Version 1.0, May 2013
Field Operations Manual
            Wadeable
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                                                                                                     75

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          National Rivers and Streams Assessment 2013/14                          Field Operations Manual
          Version 1.0, May 2013                                                            Wadeable

          Table 8.10 Procedure: Measuring Bank Characteristics
              1.   To measure bank angle, lay a meter ruler or a short (approx. 1 m long) rod down against
                  the left bank (determined as you face downstream), with one end at the water's edge. At
                  least 0.5 m of the ruler or rod should be resting comfortably on the ground to determine
                  bank angle. Lay the clinometer on the rod, and read the bank angle in degrees from the
                  external scale on the clinometer. Record the angle in the field for the left bank in the
                  BANK MEASUREMENT section of the Channel/Riparian Cross-section Form.
                  •  A vertical bank is 90°, overhanging banks have angles >90° approaching 180°, and
                     more gradually sloped banks have angles <90°. To measure bank angles >90°, turn
                     the clinometer (which only reads 0 to 90°) over and subtract the angle reading from
                     180°.
                  •  If there is a large boulder or log present at the transect, measure bank angle at a
                     nearby point where conditions are more representative.
              2.   If the bank is undercut, measure the horizontal distance of the undercutting to the
                  nearest 0.01 m. The undercut distance is the distance from the water's edge out to the
                  point where a vertical plumb line from the bank would hit the water's surface. Record
                  the distance on the field data form. Measure submerged undercuts by thrusting the rod
                  into the undercut and reading the length of the rod that is hidden by the undercutting.
              3.   Repeat Steps 1 and 2 on the right bank.
              4.   Hold the surveyor's rod vertical, with its base planted at the water's edge. Examine both
                  banks, then determine the channel incision as the height up from the water surface to
                  elevation of the first terrace of the valley floodplain (Note this is at or above the bankfull
                  channel height). Whenever possible, use the clinometer as a level (positioned so it reads
                  0% slope) to measure this height by transferring (back sighting) it onto the surveyor's
                  rod. Record this value in the INCISED HEIGHT field of the bank measurement section on the
                  field data form.
              5.   While still holding the surveyor's rod as a guide, and sighting with the clinometer as a
                  level, examine both banks to measure and record the height of bankfull flow above the
                  present water level. Look for evidence on one or both  banks such as:
                  •  An obvious slope break that differentiates the channel from a relatively flat
                     floodplain terrace higher than the channel.
                  •  A transition from exposed stream sediments to terrestrial vegetation.
 Q                •  Moss growth on rocks along the banks.
 
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                                       (A)
                 Bank Angle= Clkio meter
       Pole is lEsthg •comfortaWy-
       from wetted edge
                 Bank «ige= Udometer readng
                              Pale is "GomfortaUe"
                              from waters edge
    Not enough undercut
    exposed to detinc
    ovemantpig bank
                                            Pole rests most
                                            •comfortably -
                                            here
            (B)
                                                                 Angle— Cfawmeter readng
Too much ^>ace under
pole i measured from
water's edge
                                               Shelf is not wide enough to    ^l|i|i!;: || \;: || \;: || |j
                                                 B for determnng bank angle              .
                                                        This is as comfortable*
                                                        as pole can get from
                                                        wetted edge
                                                           Bank Angle=190* Cinometer reading
                                                                               DVP/PRK8/OS
Figure 8.9 Determining Bank Angle Under Different Types of Bank Conditions (A) typical, (B) incised
channel, (C) undercut bank, and (D) overhanging bank.
Unfortunately, we have to depend upon evidence visible during the low flow sampling season. If
available, consult published rating curves relating expected bankfull channel dimensions to
stream drainage area within the region of interest. Graphs of these rating curves can help you
get a rough idea of where to look for field evidence to determine the level of bankfull flows.
Curves such as these are available from the USGS for streams in most regions of the U.S. (e.g.,
Dunne and Leopold 1978; Harrelson et al. 1994, Leopold 1994). To use them, you need to know
the contributing drainage area to your sample site.  Interpret the expected bankfull levels from
these curves as a height above the streambed in a riffle, but remember that your field
measurement will be a height above the present water surface of the stream. Useful resources
to aid your determination of bankfull flow levels in streams in the United States are video
presentations produced by the USDA Forest Service for western streams (USDA Forest Service
1995) and eastern streams (USDA Forest Service 2002).
After consulting rating curves that show where to expect bankfull levels in a given size of
stream, estimate the  bankfull flow level by looking at the following indicators:
First look at the stream and its valley to determine the active floodplain. This is a depositional
surface that frequently is flooded and experiences sediment deposition under the current
climate and hydrological regime.
Then look specifically for:

    •   An obvious break in the slope of the banks.

    •   A change from water loving and scour tolerant vegetation to more drought tolerant
       vegetation.

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             •   A change from well sorted stream sediments to unsorted soil materials.
          In the absence of clear bankfull indications, consider the previous season's flooding as the best
          evidence available (note: you could be wrong if very large floods or prolonged droughts have
          occurred in recent years.). Look for:

             •   Drift debris ("sticky wickets" left by the previous seasons flooding).

             •   The level where deciduous leaf fall is absent on the ground (carried away by previous
                 winter flooding).

             •   Unvegetated sand, gravel or mud deposits from previous years flooding.
          In years that have experienced large floods, drift material and other recent high flow markers
          may be much higher than other bankfull indicators.  In such cases, base your determination on
          less transient indicators such as channel form, perennial vegetation, and depositional features.
          In these cases, flag your data entry and also record the height of drift material in the comments
          section of the field data form.
          We use the vertical distance (height) from the observed water surface up to the level of the first
          major valley depositional surface (Figure 8.10) as a measure of the degree of incision or
          downcutting of the stream below the general level of its valley. This value is recorded in the
          INCISED HEIGHT field. It may not be evident at the time of sampling whether the channel is
          downcutting, stable, or aggrading (raising its bed by depositing sediment). However, by
          recording incision heights measured in this way and monitoring them over time, we will be able
          to tell if streams are incising or aggrading.
          If the channel is not greatly incised, bankfull channel height and incision height will be the same
          (i.e., the first valley depositional surface is the active floodplain). However, if the channel  is
          incised greatly, the  bankfull level will be below the level of the first terrace of the valley
          floodplain, making bankfull channel height less than incision height (Figure 8.11). Bankfull height
          is never greater than incision height. You may need to look for evidence of recent flows (within
          about one year) to distinguish bankfull and incision heights. In cases where the channel is
          cutting a valley sideslope and has over-steepened and destabilized that slope, the bare
          "cutbank" against the steep hillside at the edge of the valley is not necessarily an indication  of
          recent incision. In such a case, the opposite bank may be lower, with a  more obvious terrace
          above bankfull height; choose that bank for your measurement of incised height. Examine both
          banks to more accurately determine incision height and bankfull height. Remember that incision
 ^        height is measured  as the vertical distance to the first major depositional surface above bankfull
 Q        (whether or not it is an active floodplain or a terrace. If terrace heights differ on left and right
 j^j        banks (both are above bankfull), choose the lower of the two terraces.  In many cases your
 °£        sample reach may be in a "V" shaped valley or gorge formed over eons, and the slope of the
 b        channel banks simply extends uphill indefinitely, not reaching a terrace before reaching the  top
 tx.        of a ridge. In such cases, record incision height values equal to bankfull values and make
 ^
 in        appropriate comment that no terrace is evident. Similarly, when the stream has extremely
 i-        incised into an ancient terrace, (e.g., the Colorado River in the Grand Canyon), you may crudely
 ^
 b        estimate the terrace height if it is the first one above bankfull  level. If you cannot estimate the
 <        terrace height, make appropriate comments describing the situation.

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      XV  Channel not Incised
Downcutting over
  geologic time
                                    Active
                             floodplain at or near
                             alley bottom elevation
                             (Record this height)
                           /Nlo recent incision
                               level at valley

                        _____________     4
                                                   First terrace on
                                                   valley bottom
                                                                              Second
                                                                              terrace
                                                     — banRfu
                                                    bottom
       B .
                        Channel
         Dov^ncutting over
           geologic time
 Former active floodplain
 no longer connected	
becomes new first terrace
   above banKfull level
   {Record this height),,
                                                      Former first
                                                   terrace becomes
                                                    second terrace
                                                                            ormer s
                                                                            rrace b
                                                                            third te
                                                                            • econd
                                                                            Bcomes
                                                                            -race
                                      Recent incision—
                                    banKfull level belovi/
                                    first terrace of valley
                                           bottom
Figure 8.10 Schematic Showing Relationship Between Bankfull Channel and Incision

(A) Not recently incised, and (B) recently incised into valley bottom. Note level of bankfull stage relative to
elevation of first terrace (abandoned floodplain) on valley bottom. (Stick figure included for scale).
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             A)  Deeply Incised Channel
                                  Hill Slope
                                   Incision Height (Always
                                      equal to or greater than
                                      bankfull height)

                                                 Second  Terrace

                                           First Terrace
               From Figure 7-S (Bl
r iecwd
/
P


T









^
IS
T' '
h
r— Bankfull
/ Height
/ (When
/ channel form
' is not a good
indicator, use
recent
floodina)
             B) Small stream constrained in V-shaped valley
                                          Bankfull Height
                                       (when channel form is
                                        not a good indicator
                                       use evidence of recent
                                          flooding, lack of
                                          permanent flood-
                                        intolerant vegetation)
  Flood-
  ntoferanl
  vegetation
                                       Incision He*ght=
                                       Bankful Height
                                                                        loteraul
                                                                     _, Jfegetation
  No incision:
No evidence of
  downcutting,
  vertical bank
   angle, etc.
          Figure 8.11 Determining Bankfull and Incision Heights for (A) Deeply Incised Channels, and (B) Streams
          in Deep V Shaped Valleys (Stick figure included for scale).

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8.8    Canopy Cover Measurements
Canopy cover over the stream is determined at each of the 11 cross-section transects. A
spherical densiometer (model A convex type) is used (Lemmon 1957). Mark the densiometer
with a permanent marker or tape exactly as shown in Figure 8.12 to limit the number of square
grid intersections read to 17. Densiometer readings can range from 0 (no canopy cover) to 17
(maximum canopy cover). Six measurements are obtained at each cross-section transect (four
measurements in each of four directions at mid-channel and one at each bank).
  TAPE
                         BUBBLE LEVELED"
Figure 8.12 Schematic of Modified Convex Spherical Canopy Densiometer.
From Mulvey et al. (1992). Note proper positioning with the bubble leveled and face reflected at the apex
of the "V". In this example, 10 of the 17 intersections show canopy cover, giving a densiometer reading of
10.

The procedure for obtaining canopy cover data is presented in Table 8.11. Hold the densiometer
level (using the bubble level) 0.3 m above the water surface with your face reflected just below
the apex of the taped "V", as shown in Figure 8.12. Concentrate on the 17 points of grid
intersection on the densiometer that lie within the taped "V". If the reflection of a tree or high
branch or leaf overlies any of the intersection points, that particular intersection is counted as
having cover. For each of the six measurement points, record the number of intersection points
(maximum=17) that have vegetation covering them in the CANOPY COVER MEASUREMENT section of
the Channel/Riparian Cross-section Form as shown in Figure 8.8.
Table 8.11 Procedure: Canopy Cover Measurements
 Canopy Cover Measurements with Densiometer
   1.  At each cross-section transect, stand in the stream at mid-channel and face upstream.
   2.  Hold the densiometer 0.3 m (1 ft) above the surface of the stream. Level the densiometer using the
      bubble level. Move the densiometer in front of you so your face is just below the apex of the taped
      "V".
   3.  Count the number of grid intersection points within the "V" that are covered by either a tree, a leaf,
      or a high branch. Record the value (0 to 17) in the Cf/vUp field of the canopy cover measurement
                             g

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           Canopy Cover Measurements with Densiometer
                section of the Channel/Riparian Cross-section and Thalweg Profile Form.
            4.  Face toward the left bank (left as you face downstream). Repeat Steps 2 and 3, recording the value
                in the CENL field of the field data form.
            5.  Repeat Steps 2 and 3 facing downstream, and again while facing the right bank (right as you look
                downstream). Record the values in the CfwDww and Cf/v/? fields of the field data form.
            6.  Move to the water's edge (either the left or right bank). Repeat Steps 2 and 3 again, this time facing
                the bank.  Record the value in the LEFT or RIGHT fields of the field data form. Move to the opposite
                bank and repeat.
            7.  Repeat Steps 1 through 6 at each cross-section transect (including any additional side channel
                transects established when islands are  present). Record data for each transect on a separate field
                data form.
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8.9    Visual Riparian Estimates

8.9.1  Riparian Vegetation Structure
The previous section described methods for quantifying the cover of canopy over the stream
channel. The following visual estimation procedures supplement those measurements with a
semi-quantitative evaluation of the type and amount of various types of riparian vegetation.
Riparian vegetation observations apply to the riparian area upstream 5 meters and downstream
5 meters from each of the 11 cross-section transects. They include the visible area from the
stream back a distance of 10m (~30 ft.) shoreward from both the left and right banks, creating a
10 m x 10 m riparian plot on each side of the stream (Figure 8.13). The riparian plot dimensions
are estimated, not measured. On steeply sloping channel margins, the 10 m x 10 m plot
boundaries are defined as if they were projected down from an aerial view.Table 8.12 presents
the procedure for characterizing riparian vegetation structure and composition. Figure 8.8
illustrates how measurement data are recorded on the Channel/Riparian Cross-section Form.
Conceptually divide the riparian vegetation into 3 layers: the Canopy layer (> 5 m high), the
Understory layer (0.5 to 5 m high), and the Ground cover layer (< 0.5 m high). Note that several
vegetation types (e.g., grasses or woody shrubs) can potentially occur in more than one layer.
Similarly note that some things other than vegetation are possible entries for the Ground cover
layer (e.g., barren ground).
Before estimating the areal coverage of the vegetation layers, record the type of woody
vegetation (broadleaf Deciduous, Coniferous, broadleaf Evergreen, Mixed,  or None) in each of
the two taller layers (Canopy and Understory). Consider the layer Mixed if  more than 10% of the
areal coverage is made up of the alternate vegetation type. If there is no woody vegetation in
the understory layer, record the type as None.
Estimate the areal cover separately in each of the three vegetation layers.  Note that the areal
cover can  be thought of as the amount of shadow cast by a particular layer alone when the sun
is directly  overhead. The maximum cover in each layer is 100%, so the sum of the areal covers for
the combined three layers could add up  to 300%. The four areal cover classes are Absent, Sparse
(<10%), Moderate (10 to 40%), Heavy ( 40 to 75%), and Very Heavy (>75%). These cover classes
and their corresponding codes are shown on the field data form (Figure 6.2-7). When rating
vegetation cover types for a single vegetation layer, mixtures of two or more subdominant
classes might all be given Sparse (1),  Moderate (2), or Heavy (3) ratings. One Very Heavy cover
class with  no clear subdominant class might be rated 4 with all the remaining classes rated as
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either Moderate (2), Sparse (1) or Absent (0). Note that within a given vegetation layer, two
cover types with 40-75% cover can both be rated 3, but no more than one cover type could
receive a rating of 4.
                                   10 m
              10 m
      Flow
               10 m
 RIPARIAN

    PLOT

 (Left Bank)
                                         Cross-sectibn Transect
                             5 m
          5 m
                               Instream Fish
                                 Cover Plot
 RIPARIAN

    PLOT

(Right Bank)
                                   10 m
                                                                    PRK/DVP SIOB
Figure 8.13 Riparian Zone and Instream Fish Cover Plots for a Stream Cross-Section Transect

Table 8.12 Procedure: Characterizing Riparian Vegetation Structure
 Riparian Vegetative Structure Measurements
   1.   Standing in mid-channel at a cross-section transect, estimate a 5 m distance upstream and
       downstream (10 m total length).
   2.   Facing the left bank (left as you face downstream), estimate a distance of 10 m back into the
       riparian vegetation. On steeply sloping channel margins, estimate the distance into the riparian zone
       as if it were projected down from an aerial view.
   3.   Within this 10 m x 10 m area, conceptually divide the riparian vegetation into 3 layers: a Canopy
       Layer (>5 m high), an Understory (0.5 to 5 m high), and a Ground Cover layer (<0.5 m high).
   4.   Within this 10 m x 10 m area, determine the dominant vegetation type for the CANOPY LAYER
       (vegetation  >5 m high) as; Deciduous, Coniferous, broadleaf Evergreen, Mixed, or None.  Consider
       the layer Mixed if more than 10% of the areal coverage is made up of the alternate vegetation type.
       Indicate the appropriate vegetation type in the VISUAL RIPARIAN ESTIMATES section of the
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                 Channel/Riparian Cross-section Form.
             5.   Determine separately the areal cover class of large trees (>0.3 m [1 ft] diameter at breast height
                 [dbh]) and small trees (<0.3 m dbh) within the canopy layer. Estimate areal cover as the amount of
                 shadow that would be cast by a particular layer alone if the sun were directly overhead. Record the
                 appropriate cover class on the field data form (0=absent: zero cover, l=sparse: <10%, 2=moderate:
                 10-40%, 3=heavy: 40-75%, or 4=very heavy: >75%).
             6.   Look at the UNDERSTORY layer (vegetation between 0.5 and 5 m high). Determine the dominant
                 woody vegetation type for the understory layer as described in Step 4 for the canopy layer. If there
                 is no woody vegetation in the understory layer, record the type as None.
             1.   Determine the areal cover class for woody shrubs and saplings separately from non-woody
                 vegetation within the understory, as described in Step 5 for the canopy layer.
             8.   Look at the GROUND COVER layer (vegetation <0.5 m high). Determine the areal cover class for
                 woody shrubs and seedlings, non-woody vegetation, and the amount of bare ground present as
                 described in Step 5 for large canopy trees.
             9.   Repeat Steps 1 through 8 for the right bank.
             10.  Repeat Steps 1 through 9 for all cross-section transects (including any additional side channel
                 transects established when islands are present). Uses a separate field data form at each transect.

          8.10   Instream Fish Cover, Algae, and Aquatic Macrophytes
          Over a defined area upstream and downstream of the sampling transects (Figure 8.13), crews
          shall estimate by eye  and by sounding the proportional cover of fish cover features and trophic
          level indicators including large woody debris,  rootwads and snags, brush, live trees in the wetted
          channel, undercut banks, overhanging vegetation, rock ledges, aquatic macrophytes,
          filamentous algae, and artificial structures.
          The procedure to estimate the types and amounts of instream fish cover is outlined in Table
          8.13. Data are recorded on the Channel/Riparian Cross-section Form as shown in Figure 8.8.
          Estimate the areal cover of all of the fish  cover and other listed features that are in the water
          and on the banks 5 m upstream and downstream of the cross-section (Figure 8.13). The areal
          cover  classes of fish concealment and other features are the same as those described for
          riparian vegetation (Section 8.9.1).
          The entry FILAMENTOUS ALGAE refers to long streaming algae that often occur in slow moving
          waters. AQUATIC MACROPHYTES are water loving plants, including mosses, in the stream that could
          provide cover for fish or macroinvertebrates.  If the stream channel contains live wetland
 •z_        grasses, include these as aquatic macrophytes. WOODY DEBRIS are the larger pieces of wood that
 Q        can influence cover and stream morphology (i.e., those pieces that would be included in the
 <        large woody debris tally [Section 8.5.1]). BRUSH/WOODY DEBRIS refers to smaller wood pieces that
 g£        primarily affect cover but not  morphology. LIVE TREES OR ROOTS are living trees that are within the
 (j        channel - estimate the areal cover provided by the parts of these trees or roots that are
 &.        inundated. OVERHANGING VEGETATION includes tree branches, brush, twigs, or other small debris
 ^f
 n:        that is not in the  water but is close to the stream (within 1 m of the surface) and provides
 h-        potential cover. BOULDERS are typically basketball- to car-sized particles. ARTIFICIAL STRUCTURES
 ^f
 b        include those designed for fish habitat enhancement, as well as in-channel structures that have
 <        been discarded (e.g.,  concrete, asphalt, cars, or tires) or deliberately placed for diversion,
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Table 8.13 Procedure: Estimating Instream Fish Cover
 Instream Fish Cover Measurements
   1.   Standing mid-channel at a cross-section transect, estimate a 5m distance upstream and downstream
       (10 m total length).
   2.   Examine the water and both banks within the 10 m segment of stream for the following features
       and types offish cover: filamentous algae, aquatic macrophytes, large woody debris, brush and
       small woody debris, in-channel live trees or roots, overhanging vegetation, undercut banks, boulders,
       and artificial structures.
   3.   For each cover type, estimate the areal cover. Record the appropriate cover class in the FISH
       COVER/OTHER section of the Channel/Riparian Cross-section Form:
          0=absent: zero cover,
          l=sparse: <10%,
          2=moderate: 10-40%,
          3=heavy: >40-75%, or
          4=very heavy: >75%).
   4.   Repeat Steps 1 through 3 at each cross-section transect (including any additional side channel
       transects established when islands are present). Record data from each transect on a separate field
       data form.
8.11   Human Influence
For the left and right banks at each of the 11 detailed Channel and Riparian Cross-sections,
evaluate the presence/absence and the proximity of 11 categories of human influences with the
procedure outlined in Table 8.14. Relate your observations and proximity evaluations to the
stream and riparian area within 5 m upstream and 5 m downstream from the station (Figure
8.14). Four proximity classes are used: In the stream or on the bank within 5 m upstream or
downstream of the cross-section transect, present within the 10 m x 10 m riparian plot but not
in the stream or on the bank, present outside of the riparian plot, and absent. Record data on
the Channel/Riparian Cross-section Form (Figure 8.8). If a disturbance is within more than one
proximity class, record the one that is closest to the stream (e.g., C takes precedence over P).
You may mark "P"  more than once for the same human influence observed outside of more
than one riparian observation plot (e.g. at  both Transect D and E). The rule is that you count
human disturbance items as often as you see them, BUT NOT IF you have to site through a
previously counted transect or its 10x10 meter riparian plot.
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                    C—within riparian plot
                                                                    B—on bank or in stream
                                                  P—outside plot
                                               (but do not sight through next
                                                    transect or plot)
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           Figure 8.14 Proximity Classes for Human Influences in Wadeable Streams
           Table 8.14 Procedure: Estimating Human Influence
            Estimate Human Influence
              1.  Standing mid-channel at a cross-section transect, look toward the left bank (left when facing
                 downstream), and estimate a 5 m distance upstream and downstream (10 m total length). Also,
                 estimate a distance of 10 m back into the riparian zone to define a riparian plot area.
              2.  Examine the channel, bank and riparian plot area adjacent to the defined stream segment for the
                 following human influences: (1) walls, dikes, revetments, riprap, and dams; (2) buildings; (3)
                 pavement/cleared lots (e.g., paved, gravelled, dirt parking lot, foundation); (4) roads or railroads, (5)
                 inlet or outlet pipes; (6) landfills or trash (e.g., cans, bottles, trash heaps); (7) parks or maintained
                 lawns; (8) row crops; (9) pastures, rangeland, hay fields, or evidence of livestock; (10) logging; and
                 (11) mining (including gravel mining).
              3.  For each type of influence, determine if it is present and what its proximity is to the stream and
                 riparian plot area. Consider human disturbance items as present if you can see them from the cross-
                 section transect. Do not include them if you have to sight through another transect  or its 10 m  xlO
                 m riparian plot.
              4.  For each type of influence, record the appropriate proximity class in the HUMAN INFLUENCE part of the
                 VISUAL RIPARIAN ESTIMATES section of the Channel/Riparian Cross-section Form. Proximity classes  are:
                    B (Bank)     Present within the defined 10 m stream segment and located in the stream or on the
                                stream bank.
                    C (Close)     Present within the 10 x 10 m riparian plot area.
                    P (Present)   Present, but outside  the riparian plot area.
                    0 (Absent)    Not  present within or adjacent to the 10 m stream segment or the riparian plot area
                                at the transect
              5.  Repeat Steps 1 through 4 for the right bank.
              6.  Repeat Steps 1 through 5 for each  cross-section transect, (including any additional side channel
                 transects established when islands are present). Record data for each transect on a  separate field
                 form.
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8.12   Cross-section Transects on Side Channels
If the wetted channel is split by an island, and the estimated flow in the side channel is less than
or equal to 15% of the total flow, the bank and riparian measurements are made at each side of
the main channel (the minor side channel is ignored other than to note its presence on the
Thalweg Profile Form), so one riparian plot is established on the island as shown in Figure 8.15
(side A). If an island is present that creates a major side channel containing more than 15% of
the total flow, an additional cross-section transect is established for the side channel as shown
in  Figure 8.15 (side B). Separate substrate, bank and riparian measurements are made for side
channel transects. Data from the additional side channel transect are recorded on a separate
Channel/Riparian Cross-section Form as shown in Figure 8.16. Riparian plots established on the
island for each transect may overlap (and be < 10 m shoreward) if the island is less than 10 m
wide at the transect.
    A) Island and minor side channel
     <15% total flow occurs in side channel
 •   No side channel cross section transect
 •   Note presence on field form
 •   Riparian plot established on island
                       10m
  I Flow
r^
y^
l(
^ 	 — -
n cross-section
isect (e.g., E)
" >
~v




RIPARIAN
PLOT
(Left Bank)
"T"""
i
— i
5m ' 5m
Instream Fish
Cover Plot
1
1
RIPARIAN
PLOT
(Right Bank)
	


10m
^^— ••





10

                      10m
B) Island and major side channel
 >15% total flow occurs in side channel
Side channel cross-section must be assessed
Two riparian plotsestablished on island (mayoverlap)
Data is recorded on additional PHab form
Figure 8.15 Riparian and Instream Fish Cover Plots for a Stream with Minor and Major Side Channels
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00
00
PHYSICAL HABITAT CHARACTERIZATION
1
00
01
n
annel/Riparian Cross-Section Form for an Additional Major Side Channel Transect.



0 NRSA 2013714 PHAB: CHANNEL/RIPARIAN CROSS-SECTION - WADEABLE ONLY •
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• Flag codes: K - Sample not collected; U = Suspect sample; F1, F2, etc. * flag assigned by field crew. Explain all flags m comment sections. 998 962 51 8*7
' 03/06/201 3 201 3 Phab Cha rtnel Rtparian - Watfeable

\lational Rivers and Streams Assessment 2013/14 Field Operat
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8.13  Channel Constraint, Debris Torrents, Recent Floods, and Discharge

8.13.1 Channel Constraint
After completing the thalweg profile and riparian/channel cross-section measurements and
observations, envision the stream at bankfull flow and evaluate the degree, extent and type of
channel constraint, using the procedures presented in Table 8.15. Record data on the Channel
Constraint Assessment Form (Figure 8.17). First, classify the stream reach channel pattern as
predominantly a single channel, an anastomosing channel, or a braided channel (Figure 8.18):
    1.  Single channels may have occasional in-channel bars or islands with side channels, but
       feature a predominant single channel, or a dominant main channel with a subordinate
       side channel.
    2.  Anastomosing channels have relatively long major and minor channels (but no
       predominant channel) in  a complex  network, diverging and converging around many
       vegetated islands. Complex channel  pattern remains even during major floods.
    3.  Braided channels also have multiple branching and rejoining channels, (but no
       predominant channel) separated by unvegetated bars. Channels are generally smaller,
       shorter, and more numerous, often  with no obvious dominant channel. During major
       floods, a single continuous channel may develop
After classifying the channel pattern, determine whether the channel is constrained within a
narrow valley, constrained by local features  within a broad valley, unconstrained and free to
move about within a broad floodplain, or free to move about, but within a relatively narrow
valley floor. Then examine the channel to ascertain the bank and valley features that constrain
the stream. Entry choices for the type of constraining features are bedrock, hillslopes,
terraces/alluvial fans, and human land use (e.g., a road, a dike, landfill, rip-rap, etc.). Estimate
the percent of the channel margin in contact with constraining features (for unconstrained
channels, this is 0%). To aid in this estimate, you may wish to refer to the individual transect
assessments of incision and constraint. Finally, estimate the "typical" bankfull channel width and
estimate the average width of the valley floor either with a topographic map or visually. If you
cannot directly estimate the valley width (e.g., it is further than you can see, or  if your view is
blocked by vegetation), record the distance you can see and mark the appropriate bubble on the
field form.
Table 8.15 Procedure: Assessing Channel Constraint
 Channel Constraint
 NOTE: These activities are conducted after completing the thalweg profile and littoral-riparian
 measurements and observations, and represent an evaluation of the entire stream reach.
 CHANNEL CONSTRAINT: Determine the degree, extent, and type of channel constraint based on envisioning the
 stream at bankfull flow.
   1.   Classify the stream reach channel pattern as predominantly a single channel, an anastomosing
       channel, or a braided channel.
       •  Single channels may have occasional in-channel bars or islands with side channels, but feature a
          predominant single channel, or a dominant main channel with a subordinate side channel.
       •  Anastomosing channels have relatively long major and minor channels branching and rejoining
          in a complex network separated by vegetated islands, with no obvious dominant channel.
       •  Braided channels also have multiple branching and rejoining channels, separated by
          unvegetated bars. Subchannels are generally small, short, and numerous, often with no obvious
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            Channel Constraint
              2.
    dominant channel.
After classifying the channel pattern, determine whether the channel is constrained within a narrow
                  valley, constrained by local features within a broad valley, unconstrained and free to move about
                  within a broad floodplain, or free to move about, but within a relatively narrow valley floor.
              3.   Then examine the channel to ascertain the bank and valley features that constrain the stream. Entry
                  choices for the type of constraining features are bedrock, hillslopes, terraces/alluvial fans, and
                  human land use (e.g., a road, a dike, landfill, rip-rap, etc.).
              4.   Based on your determinations from Steps 1 through 3, select and record one of the constraint
                  classes shown on the Channel Constraint Form.
              5.   Estimate the percent of the channel margin in contact with constraining features (for unconstrained
                  channels, this is 0%). Record this value on the Channel Constraint Form.
              6.   Finally, estimate the "typical" bankfull channel width, and visually estimate the average width of the
                  valley floor. Record these values on the Channel Constraint Form.
            NOTE: To aid in this estimate, you may wish to refer to the individual transect assessments of incision and
            constraint that were recorded on the Channel/Riparian Cross-Section Forms.
            NOTE: If the valley is wider than you can directly estimate, record the distance you can see and mark the
            bubble on the field form.
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     Site ID:
                                         NRSA 2013/14 CHANNEL CONSTRAINT

                                                                    Date:         /
      CHANNEL PATTERN (Fill in one):
     O One Channel

     O Anastomosing (complex) channel - (Relatively long major and minor channels branching and rejoining.)

     O Braided channel - (Multiple short channels branching and rejoining - mainly one channel broken up by
         numerous mid-channel bars.)
      CHANNEL CONSTRAINT)Fill in one):
     O Channel very constrained in V-shaped valley (i.e. it is very unlikely to spread out over valley or erode a
          new channel during flood)
     O Channel is in Broad Valley but channel movement by erosion during floods is constrained by Incision
         (Flood flows do not commonly spread over valley floor or into multiple channels.)
     O Channel is in Narrow Valley but is not very constrained, but limited in movement by relatively narrow
         valley floor (< -10 x bankfull width)
     O Channel is Unconstrained in Broad Valley (i.e. during flood it can Fill off-channel areas and side channels,
         spread out over flood plain, or easily cut new channels by erosion)
      CONSTRAINING FEATURES (Fill in one):
     O Bedrock (i.e channel is a bedrock-dominated gorge)

     O Hillslope (i.e. channel constrained in narrow V-shaped valley)

     O Terrace (i.e channel is constrained by its own incision into river/stream gravel/soil deposits)

     O Human Bank Alterations (i.e. constrained by rip-rap, landfill, dike, road, etc.)

     O No constraining features
      Percent of channel length with margin
        in contact with constraining feature: i_
                                             (0-100% |
      Bankfull width:
                                (m)
                                                                              Percent of Channel Margin Examples
                                                                                 100%
                                                                                                         100%
Valley width (Visual Estimated Average):               (m)

Note: Be sure to include distances between both sides of valley border for valley witflh.
If you cannot see the valley borders, record the distance  /-\
you can see and fill this bubble.
                                                                                50%
                                                                                                        50%
      COMMENTS
              04/06/2013 2013 Channel Constraint
                                                                                                    1742268066
Figure 8.17 Channel Constraint Form
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            A)  Anastomosing channel pattern
                  Vegetated islands above bankfull flow.  Multiple
                  channels remain during  major flood events.
             B) Braided channel pattern

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         Unvegetated bars below bankfull flow.  Multiple
         channel pattern disappears during major flood events.
                                                           DVP
Figure 8.18 Types of Multiple Channel Patterns

8.13.2 Debris Torrents and Recent Major Floods
Debris torrents, or lahars, differ from conventional floods in that they are flood waves of higher
magnitude and shorter duration, and their flow consists of a dense mixture of water and debris.
Their high flows of dense material exert tremendous scouring forces on streambeds. For
example, in the Pacific Northwest, flood waves from debris torrents can exceed 5 meters deep
in small streams normally 3 m wide and 15  cm deep. These torrents move boulders in excess of
1 m diameter and logs >1 m diameter and >10 m long. In temperate regions, debris torrents
occur primarily in steep drainages and are relatively infrequent, occurring typically less than
once in several centuries.
Because they may alter habitat and biota substantially, infrequent major floods and torrents can
confuse the interpretation of measurements of stream biota and habitat in regional surveys and
monitoring programs. Therefore, it is important to determine if a debris torrent or major flood
has occurred within the recent past. After completing the thalweg profile and channel/riparian
measurements and observations, examine the stream channel along the entire sample reach,
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including its substrate, banks, and riparian corridor, checking the presence of features described
on the Torrent Evidence Assessment Form (Figure 8.19). It may be advantageous to look at the
channel upstream and downstream of the actual sample reach to look for areas of torrent scour
and massive deposition to answer some of the questions on the field form. For example, you
may more clearly recognize the sample reach as a torrent deposition area if you find extensive
channel scouring upstream. Conversely, you may more clearly recognize the sample reach as a
torrent scour reach if you see massive deposits of sediment, logs, and other debris downstream.
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                    Site ID:
                                               NRSA 2013/14 TORRENT EVIDENCE ASSESSMENT

                                                                                 Date:        /         /
                                                                                                           R»vi*w«dby(ln*Gil>:_
                   Please fill in any of the following that are evident.

                   EVIDENCE OF TORRENT SCOURING:
                     O
                     O
              01 - Stream channel has a recently devegetated corridor two or more times the width of the low flow channel. This
              corridor lacks riparian vegetation with possible exception of fireweed, even-aged alder or cottonwood seedlings, grasses.
              or other herbaceous plants.
                            02 - Stream substrate cobbles or large gravel particles are NOT IMBRICATED.  (Imbricated means that they lie with flat
                            sides horizontal and that they are stacked like roof shingles -- imagine the upstream direction as the top of the "roof.") In
                            a torrent scour or deposition channel, the stones are laying in unorganized patterns, lying "every which way."  In addition
                            many of the substrate particles are angular (not "water-worn.")
                            03 - Channel has little evidence of pool-riffle structure. {For example, could you ride a mountain bike down the channel?)
                            04 - The stream channel is scoured down to bedrock for substantial portion of reach.
                              - There are gravel or cobble berms (little levees) above bankroll level.
                            06 - Downstream of the scoured reach (possibly several miles), there are massive deposits of sediment, logs, and other
                            debris.
              07 - Riparian trees have fresh bark scars at many points along the stream at seemingly unbelievable heights above the
              channel bed.
                            08 - Riparian trees have fallen into the channel as a result of scouring near their roots.
                   EVIDENCE OF TORRENT DEPOSITS:
                     O
                     O
              OS - There are massive deposits of sediment, logs, and other debris in the reach. They may contain wood and boulders
              that, in your judgement, could not have been moved by the stream at even extreme flood stage.
              10 - If the stream has begun to erode newly laid deposits, it is evident that these deposits are "MATRIX SUPPORTED."
              This means that the large particles, like boulders and cobbles, are often not touching each other, but have silt, sand, and
              other fine particles between them (their weight is supported by these fine particles — in contrast to a normal stream
              deposit, where fines, if present, normally "fill-in" the interstices between coarser particles.)
                   NO EVIDENCE:
                     O
              11 - No evidence of torrent scouring or torrent deposits.
                   COMMENTS
                        01/08/2013  2013 Torrent Evidence
                                                                                                                3362134147
Figure 8.19 Torrent Evidence Assessment Form
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8.14  Stream Discharge
No single method for measuring discharge is applicable to all types of stream channels. The
preferred procedure for obtaining discharge data is based on "velocity-area" methods (e.g.,
Rantz and others, 1982; Linsley et al., 1982). For streams that are too small or too shallow to use
the equipment required for the velocity-area procedure, two alternative procedures are
presented. One procedure is based on timing the filling of a volume of water in a calibrated
bucket. The second procedure is based on timing the movement of a neutrally buoyant object
(e.g., an orange or a small rubber ball) through a measured length of the channel, after
measuring one or more cross-sectional depth profiles within that length.

8.14.1 Velocity-Area Procedure
Because velocity and depth typically vary greatly across a stream, accuracy in field
measurements is achieved by measuring the mean velocity and cross-sectional area of many
increments across a channel (Figure 8.20). Each increment gives a subtotal of the stream
discharge, and the whole is calculated as the sum of these parts. Discharge measurements are
made at only one carefully chosen channel cross-section within the sampling reach. It is
important to choose a channel cross-section that is as much like a canal as possible. A glide area
with  a "U" shaped channel cross-section that is free of obstructions provides the best conditions
for measuring discharge by the velocity-area method. You may remove rocks and other
obstructions to improve the cross-section  before any measurements are made. However,
because removing obstacles from one part of a cross-section affects adjacent water velocities,
you must not change the cross-section once you commence collecting the set of velocity and
depth measurements.
The procedure for obtaining depth and velocity measurements is outlined in
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           Table 8.16. Record the data from each measurement on the Stream Discharge Form as shown in
           Figure 8.21. In the field, data will be recorded using only one of the available procedures.
                    15 to 20 equally spaced
                    intervals across stream.
                    beginning at left margin
Measure stream depth at the midpoint
of each interval, and obtain velocity
measurements at 0.6 depth
Extended surveyor's
rod or tape measure
                                                                             \
Record distance
and depth of
right margin
           Figure 8.20 Layout of Channel Cross-Section for Obtaining Discharge Data by the Velocity-Area
           Procedure
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Table 8.16 Procedure: Determining Stream Discharge - Velocity-Area
 Stream Discharge Velocity- Area Option
 1.  Locate a cross-section of the stream channel for discharge determination that has most of the
     following qualities (based on Rantz and others, 1982):
     •   Segment of stream above and below cross-section is straight
     •   Depths mostly greater than 15 centimeters, and velocities mostly greater than 0.15
         meters/second. Do not measure discharge in a pool.
     •   "U" shaped, with a uniform streambed free of large boulders, woody debris or brush, and dense
         aquatic vegetation.
     •   Flow is relatively uniform, with no eddies, backwaters, or excessive turbulence.
 2.  Lay the surveyor's rod (or stretch a measuring tape) across the stream perpendicular to its flow, with
     the "zero" end of the rod or tape on the left bank, as viewed when looking downstream. Leave the
     tape tightly suspended across the stream, approximately one foot above water level.
 3.  Attach the velocity meter probe to the calibrated  wading rod. Check to ensure the meter is functioning
     properly and the correct calibration value is displayed. Calibrate (or check the calibration) the velocity
     meter and probe as directed in the meter's operating manual. Darken in the "VELOCITY AREA" bubble
     on the Stream  Discharge Form.
 4.  Divide the total wetted stream width into 15 to 20 equal sized intervals. To determine interval width,
     divide the width by 20 and round up to a convenient number. Intervals should not be less than 10 cm
     wide, even if this results in less than 15 intervals. The first interval is located at the left margin of the
     stream (left when  looking downstream), and the last interval is located at the right margin of the
     stream (right when looking downstream).
 5.  Stand downstream of the rod or tape and to the side of the first interval point (closest to the left bank
     if looking downstream).
 6.  Place the wading rod in the stream at the interval point and adjust the probe or propeller so that it is
     at the water surface. Darken in the appropriate bubbles for "Distance Units" and  "Depth Units" on the
     Stream Discharge Form. Record the distance from the left bank and the depth indicated on the wading
     rod on the Stream Discharge Form.
     Note: For the first  interval, distance equals 0 cm, and in many cases depth may also equal 0 cm. For
     the last interval, distance will equal the wetted width (in cm) and depth may again equal 0 cm.
 7.  Stand downstream of the probe or propeller to avoid disrupting the stream flow. Adjust the  position
     of the probe on the wading rod so it is at 0.6 of the measured depth below the surface of the water.
     Face the probe upstream at a right angle to the cross-section, even if local flow eddies hit at oblique
     angles to the cross-section.
 8.  Wait 20 seconds to allow the meter to equilibrate, then measure the velocity. Darken the appropriate
     "Velocity Units" bubble on the Stream  Discharge Form. Record the value on the Stream Discharge
     Form. Note for the first interval, velocity may equal 0 because depth will equal 0.
     •   For the electromagnetic current meter (e.g., Marsh-McBirney), use the lowest time constant scale
         setting on  the meter that provides stable readings.
     •   For the impeller type meter (e.g., Swoffer 2100), set the  control knob at the mid-position of
         "DISPLAY AVERAGING". Press "RESET" then "START" and proceed with the measurements.
 9.  Move to the next interval point and repeat Steps 6 through 8. Continue until depth and velocity
     measurements have been recorded for all intervals. Note for the last interval (right margin),  depth and
     velocity values may equal 0.
     At the last interval (right margin), record a "Z" flag on the field form to denote the last interval
     sampled.
     If using a meter that computes discharge directly, darken the "Q" bubble on the discharge form, and
     record calculated discharge value. In this case, you do not have to record the depth and velocity data
     for each interval.
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NRSA 2013/14 DISCHARGE - WADEABLE ONLY
Date:


O Velocity Area
Distance Units
O ft O cm
Depth Units
Of O cm
Velocity Units
OffaXX.X Om'sX.XX
Dist from Bank Depth Velocity Flag
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
0



















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Flag









































































R»vi«w«d bv [irr-ul : ^B
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OTimed Filling
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1
2
3
4
5
Volume (L)





Time(s)





Flag






O Neutral Buoyant Object

Float Olst.
O ft O m
Float TlmB
(s)
Flag
Float 1



Float 2



Float 3



Cross Sections on Float Reach

Width
Oft Om
Depth 1
Of O cm
Depth 2
Depth 3
Depth .!
Depth ',
Upper Section
. • | 	








Middle Section
. • ( 	








Lower Section
. • t 	 t








if discharge is determined directly ,
in Held record value here: Q= °cfs Om/s FLAG
Comments



I CM/OS/2013 2013 D scharee 8 10304 893 6 |
Figure 8.21 Discharge Form, showing data recorded for All Discharge Measurement Procedures
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8.14.2 Timed Filling Procedure
In channels too "small" for the velocity-area method, discharge can sometimes be measured by
filling a container of known volume and timing the duration to fill the container. "Small" is
defined as a channel so shallow that the current velocity probe cannot be placed in a cross-
section of the stream that contains one or more natural spillways or plunges that  collectively
include the entire stream flow, or where the channel is broken up and irregular due to rocks and
debris, and a suitable cross-section for using the velocity area procedure is not available. . This
can be an extremely precise and accurate method, but requires a natural or constructed spillway
of freefalling water. A temporary spillway can also be constructed using a portable V-notch weir,
plastic sheeting, or other materials that are available onsite. If obtaining data by this procedure
will result in a lot of channel disturbance or stir up a  lot of sediment, wait until after all biological
and chemical measurements and sampling activities  have been completed.
Choose a location within the sampling reach that is narrow and easy to block when using a
portable weir. Position the weir in the channel so that the entire flow of the stream is
completely rerouted through its notch (Figure 8.22). Impound the flow with the weir, making
sure that water is not flowing beneath or around the side of the weir. Use mud or stones and
plastic sheeting to get a good waterproof seal. The notch must be high enough to create a small
spillway as water flows over its sharp crest.
                              Water Level
                                                                      Bucket
Figure 8.22 Use of a Portable Weir in Conjunction with a Calibrated Bucket to Obtain an Estimate of
Stream Discharge

The timed filling procedure is presented in Table 8.17. Make sure that the entire flow of the
spillway is going into the bucket. Record the time it takes to fill a measured volume on the
Discharge Measurement Form as shown in Figure 8.21. Repeat the procedure 5 times. If the
cross-section contains multiple spillways, you will need to do separate determinations for each
spillway. If so, clearly indicate which time and volume data  replicates should be averaged
together for each spillway; use additional Stream Discharge Form if necessary.
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          Table 8.17 Procedure: Determining Stream Discharge - Timed Filling
            Stream Discharge Timed filled Option
            NOTE: If measuring discharge by this procedure will result in significant channel disturbance or will stir up
            sediment, delay determining discharge until all biological and chemical measurement and sampling
            activities have been completed.
            1.  Choose a cross-section that contains one or more natural spillways or plunges, or construct a
               temporary one using on-site materials, or install a portable weir using a plastic sheet and on-site
               materials.
            2.  Darken the 'TIMED FILLING" bubble in the stream discharge section of the Stream Discharge Form.
            3.  Position a calibrated bucket or other container beneath the spillway to capture the entire flow. Use a
               stopwatch to determine the time required to collect a known volume of water. Record the volume
               collected (in liters) and the time required (in seconds) on the Stream Discharge Form.
            4.  Repeat Step 3 a total of 5 times for each spillway that occurs in the cross-section. If there is more
               than one spillway in a cross-section, you must use the timed filling approach on all of them.
               Additional spillways may require additional data forms
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          8.14.3 Neutrally Buoyant Object Procedure
          In very small, shallow streams with no waterfalls, where the standard velocity-area or timed
          filling methods cannot be applied; the neutrally buoyant object method may be the only way to
          obtain an estimate of discharge. The required pieces of information are the mean flow velocity
          in the channel and the cross-sectional area of the flow. The mean velocity is estimated by
          measuring the time it takes for a neutrally buoyant object to flow through a measured length of
          the channel. The channel cross-sectional area is determined from a series of depth
          measurements along one or more channel  cross-sections. Since the discharge is the product of
          mean velocity and channel cross-sectional area, this method is conceptually very similar to the
          standard velocity-area method.
          The neutrally buoyant object procedure is described  in Table 8.18. Examples of suitable objects
          include plastic golf balls (with holes), small  sponge rubber balls, or small sticks. The object must
          float, but very low in the water. It should also be small enough that it does not "run aground" or
          drag bottom. Choose a stream segment that is roughly uniform in cross-section, and that is long
          enough to require 10 to 30 seconds for an object to float through it. Select one to three cross-
          sections to represent the channel dimensions within  the segment, depending on the variability
          of width and/or depth. Determine the stream depth at 5 equally spaced points at each cross-
          section. Three separate times, measure the time required for the object to pass through the
          segment that includes all of the selected cross-sections. Record data on the Stream Discharge
          Form as shown in Figure 8.21.
          Table 8.18 Procedure: Determining Stream Discharge - Neutrally Buoyant Object
            Stream Discharge Neutrally Buoyant Object Option
1.  Darken the "NEUTRALLY BUOYANT OBJECT" bubble on the Stream Discharge Form.
   Select a segment of the sampling reach that is deep enough to float the object freely and long enough
   that it will take between 10 and 30 seconds for the object to travel. Mark the units used and record
   the length of the segment in the "FLOAT DIST." field of the Stream Discharge Form.
   If the channel width and/or depth change substantially within the segment, measure widths and
   depths at three cross-sections, one near the upstream end of the segment, a second near the middle
   of the segment, and a third near the downstream end of the segment.
   If there is little change in channel width and/or depth, obtain depths from a single "typical" cross-
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    section within the segment.
 4.  At each cross-section, measure the wetted width using a surveyor's rod or tape measure, and record
    both the units and the measured width on the Stream Discharge Form. Measure the stream depth
    using a wading rod or meter stick at points approximately equal to the following proportions of the
    total width: 0.1,0.3, 0.5, 0.7, and 0.9. Record the units and the depth values (not the distances) on the
    Stream Discharge Form.
 5.  Repeat Step 4 for the remaining cross-sections.
 6.  Use a stopwatch to determine the time required for the object to travel through the segment. Record
    the time in the "FLOATTIME" field of the Stream Discharge Form.
 7.  Repeat Step 6 two more times. The float time may differ somewhat for the three trials.
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          9      FECAL INDICATOR (ENTEROCOCCI)
          9.1     Summary of Method
          Collect a fecal indicator sample at the last transect (Transect K) after all other sampling is
          completed. Filters must be frozen within 6 hours of collection. Use a pre-sterilized, 250 ml
          bottle and collect the sample approximately 1 m off the bank at about 0.3 meter (12 inches)
          below the water. Following collection, place the sample in a cooler and maintain on ice prior to
          filtration of two 50 ml volumes. Again, samples must be filtered and frozen on dry ice within 6
          hours of collection. In addition to collecting the sample, look for signs of disturbance throughout
          the reach that would contribute to the presence of fecal contamination to the waterbody.
          Record these disturbances on the Site Assessment Form (Error! Reference source not found.).
          9.2     Equipment and Supplies
          Table 9.1 provides the equipment and supplies  needed to collect the fecal indicator sample.
          Record the sample data on the Sample Collection Form (Error! Reference source not found.).
          Table 9.1 Equipment and Supplies: Fecal Indicator Sampling (Wadeable Sites)
           For collecting samples
                            nitrile gloves
                            pre-sterilized, 250 ml sample bottle
                            sodium thiosulfate tablet
                            Wet ice
                            cooler
           For recording measurements
                           Sample Collection Form
                           Fecal Indicator sample labels (2 vial labels and 1 bag label)
                           Pencils (for data forms)
                           Fine tipped indelible markers (for labels)
                           Clear tape strips
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          9.3     Sampling Procedure
          The procedure for collecting the fecal indicator sample is presented in Table 9.2.
          Table 9.2 Procedure: Fecal Indicator (Enterococci) Sample Collection (Wadeable Sites)
           Enterococci Sample
1.   Put on sterile, nitrile gloves.
2.   Select a sampling location at transect K that is approximately 1 m from the bank and approximately
    0.3m deep. Approach the sampling location slowly from downstream or downwind.
3.   Lower the uncapped, inverted 250 ml sample bottle to a depth of 1 foot below the water surface,
    avoiding surface scum, vegetation, and substrates.
4.   Point the mouth of the container away from the body or boat. Right the bottle and raise it through
    the water column, allowing bottle to fill completely.
5.   If the depth does not reach 0.3m along transect at 1 m from the bank, take the sample and flag it on
    the field form.
6.   After removing the container from the water, discard a small portion of the sample to allow for
    proper mixing before filtering.
7.   Add the sodium thiosulfate tablet, cap, and shake bottle 25 times.
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   8.   Store the sample in a cooler on ice to chill (do not freeze immediately). Chill for at least 15 minutes.
   9.   Sample must be filtered and all filters frozen within 6 hours of collection
9.4     Sample Processing in the Field
You will need to process two separate filters for the Enterococci sample. All the filters required
for an individual site should be sealed in plastic bags until use to avoid external sources of
contamination. Please refer to Section 13.3 for information regarding processing the Enterococci
samples.
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          10     FISH ASSEMBLAGE

          10.1   Summary of Method
          The fish sampling method is designed to provide a representative sample of the fish community,
          collecting all but the rarest fish taxa inhabiting the site. It is intended to accurately represent
          species richness, species guilds, relative abundance, size, and presence of anomalies. The
          intended uses of the fish assemblage data are to calculate predictive models of multimetric
          indicators (MMIs; similar to an Index of Biotic Integrity (IBI); Pont et al. 2008, USEPA 2013a) and
          possibly Observed/Expected (O/E) taxa richness. In addition, the fish assemblage data provides a
          starting point for developing potential indicators of ecosystem services related to fish.
          For wadeable streams, collect fish using a bank or towed barge electrofishing unit (2,500- to
          5,000-V; 2.5-5.0 GPP unit or equivalent). Use a backpack electrofishing unit in smaller streams,
          when conductivity is appropriate. As a last option, use seining as an alternate method when
          electrofishing is precluded by high conductivity or extreme levels of turbidity.
          There are different protocols for collecting fish from wadeable streams of different sizes (see
          Section  10.3). At smaller streams (<12.5 m wide), sample all available habitats over the entire
          support reach (40 channel widths [CW]). At  medium and large wadeable streams (>12.5 m
          wide), the initial  length of the fish sampling  reach is less than the entire sampling reach, and
          effort is focused  on habitats along the stream margins. For all wadeable sites, conduct sampling
          in an upstream direction (i.e., from the downstream end of the reach), allocating effort (button
          time) within subreaches (areas between the cross-section transects). At medium and large
          wadeable streams, if you have not collected 500 individuals at the end of the defined fish
          sampling reach, sample additional subreaches until you obtain at least 500 individuals or until
          you have sampled the entire reach.  Record  information related to sampling effort on the front
          of the Fish Gear and Sample Information Form (Figure 10.1). Record species identification and
          enumeration data on one or more pages of  the Fish Collection Form (Figure 10.2). There are
          several revisions and clarifications to the wadeable sampling protocols from those used in the
          NRSA 2008/09.
          The layout of the fish sampling reach and how to allocate effort across the reach for smaller and
          larger wadeable  streams is clarified. The Fish Gear Form is modified to include the specific
          protocol used to sample (wadeable or large wadeable), to clarify reasons for not sampling. The
          collection form is modified to include information about introduced species and hybrids and to
          break down total counts by major size classes (to begin to look at ecosystem service-related
          indicators). Measuring minimum and maximum lengths for each species collected is not
          required. An additional form to record seining effort information is used to evaluate the
          sufficiency of the seining results and the ability to combine them with electrofishing data. The
          procedure for preparing voucher specimens is clarified to distinguish unknown/range extension
          vouchers (those taken back to a local facility for identification) and  quality assurance (QA)
          vouchers (a sample of all species from a site sent to an external facility for confirmation of all
          field identifications). Guidance for recording unknown taxa is clarified to minimize ambiguity.
          The procedure to update species identifications based on the results from unknown/range
          extension voucher specimens is clarified and uses an additional data form.
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Site ID:
FISH SAMPLING PROTOCOL (select one):
O BOATABLE(20to40ChannelWidtr!S(CW)>
O L<3- WADEABLE - (>=12.5m wide) (500 m to 40 C\
O WADEABLE - (<12.5 m wide) (40 CW)
Final Length of
Fishing Reach (m):
O Fast flowing high gradient site Sampli
FISH GEAR INFORMATION
WaterVisibility: QGood OPoor
Primary Electrofishing Gear
# of Netters(1):
Anodes
O RAFT (No motor) Number.
O BACKPACK
Diameters:
O BANK OR TOWED BARGE
Secondary Electrofishing Gear
*ofNetters(1):
Anodes
O RAFT (No motor) Number
O BACKPACK
Diameters:
O BANK OR TOWED BARGE

•ORMAI ION (Front) BT
Date: / /
FISH SAMPLING - NOT CONDUCTED OR SUSPENDED (select one):
O Fished - None Collected
vi O Not Fished/Fishing suspended - Can't sample >/= 50% of
required reach:
- Boatable (1 0 CW)
• Lg. Wadeable (250m or 10 CW, whichever is longer)
- Wadeable (20 CW)
ng Protocol Comments:
O Not Fished - No Permit
O Not Fished - Site Conditions Prohibit Sampling (Describe in comments)
O Fishing Suspended - Permit Restriction (Listed species encountered)
O Not Fished - Equipment Failure
O Not Fished - Other Explain:


Water Temp(°C): Cond(uSfern):
Wave Form: Volts: (50-1000)
f\ A/~ .» ii .«« 1 L. >
_ _ _ 2500 o r 5000 ( boatfra ft) ,_

Q in. O Pulsed DC pps or H2
provided for bp) _
Wave Form: Volts: (50-1000)
f\ Ar~
rt__ 2500 or 5000 (boat/raft) .

O in. O Pulsed DC pps or Hi
provided for bp)

Pulse Width (ms):
Total Shock(button) Time (s):
Total FishlngTime(min):
% of Fishing Reach Sampled:
Pulse Width(ms):
Total Shock(button) Time (s):
Total FishingTime(min):
% of Fishing Reach Sampled:
Primary Seine Net: QSAGSBNE Q MINNOW SEINE No. of crew members:
Height(m): Length(m): Mesh(mm]: va. 3U H^- °f
?m*m!nS 	 % of Fishing Reach Sampled:
Secondary Seine Net: O BAG SEINE O MINNOW SEINE No. of crew members:
Height(m): Length(m): Mesh(mm): Lenoih^ml' Hauls' T°T
im^min" 	 % of Fishing Reach Sampled:
GEAR INFORMATION COMMENTS


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                         Wadeable
10.2  Equipment and Supplies
Table 10.1 shows the checklist of equipment and supplies required to complete the fish
assessment. This checklist is similar to the one presented in Appendix A, which is used at the
base location to ensure that all of the required equipment is brought to the site.
Table 10.1 Equipment and Supplies: Fish Collection (Wadeable Sites)
 For collecting       Bank or towed barge electrofishing
 samples           equipment (including variable voltage
                   pulsator unit, wiring cables, generator,
                   and electrodes)
                   Backpack electrofishing unit with anode
                   and cathode
                   Linesman gloves, boots, and other
                   necessary safety equipment
                   Dip nets
                   Extra electrofishing unit batteries
                   Scientific collection permit(s)
                   Digital camera with extra memory card &
                   battery
                   1 Laser rangefinder (optional)
                   Polarized sunglasses and hats
                   10 ft x 6 ft Minnow or Bag Seine with %
                   inch mesh (additional 4' depth seine may
                   also be used)
 For recording       Sheet of sample labels and voucher
 measurements      specimen tags (for unknown/range
                   extension voucher samples)
                   Sheet of sample labels and voucher
                   specimen tags (for QA voucher samples)
                   Fish Gear Form
                   Fish Collection forms
1 Scalpel for slitting open large fish
before preservation.
1 container of 10% buffered formalin
Small mesh bags or several Leak proof
HOPE jars (various sizes from 250 mL-4
L) for fish voucher specimens
2 non-conducting dip nets with % inch
mesh
1 Minnow net for dipping small fish from
live well
2 measuring boards (3 cm size classes)
Iset Fish ID keys
Field Operations Manual and/or
laminated Quick Reference Guide
20 ft x 6 ft Minnow or Bag Seine with %
inch mesh (additional 4 ft depth seine
may also be used)

Clear tape strips
Soft (#2) lead pencils
Fine-tip indelible markers
10.3  Sampling Procedures
The reach length sampled for fish varies based on the width of the stream used to establish the
length of the support reach and on the number of individuals collected (Figure 10.3). For small
wadeable streams (with a mean channel width [CW] < 12.5 meters), follow the protocol
presented in Section 0. Sample the entire support reach (150 to 500 m) and move the
electrofishing unit (towed barge or backpack) within each subreach to sample both shorelines as
well as the mid-channel.
For medium and large wadeable streams (mean CW > 12.5  m), follow the protocols described in
Section 10.3.3. In medium wadeable streams (mean CW between 12.5 and 25 meters) the
minimum length for fish sampling is 500 m, which will include between 5 and 10 subreaches. If
you reach the required distance within a subreach, extend the length of the fish sampling reach
to the end of the subreach so you end fishing efforts at a transect. For large wadeable stream
(mean CW > 25 meters), the minimum length for fish sampling is 5 of the 10 subreaches. In both
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           medium and large wadeable streams, if a minimum of 500 fish are not collected after sampling
           the minimum fishing reach,  additional subreaches will be sampled until 500 fish are collected or
           all 10 subreaches have been sampled. Table 10.2 summarizes the fishing protocols for each of
           three sizes of wadeable streams.
           If conditions prohibit any type of electrofishing, collect fish by seining as described in Table 10.3.
           The objective of seining is to collect species and relative abundance data that is comparable to
           what would have been obtained by electrofishing at the site.

           10.3.1 Irruptive Species
           For the purposes of NRSA, the term irruptive species will be used to describe fish species which
           are found in locally abundant "patches" in one or two small places within the sampling reach.
           These  are distinct from dominant species which are in abundance throughout most of the reach.
           As such, irruptive species  may artificially skew necessary effort to reach 500 individuals; and, if
           included the overall assemblage counts, may artificially skew the calculations of relative
           abundance offish species in the reach. To avoid the impact of irruptive species, move quickly
           through large isolated schools of a single species (e.g., shad, certain shiners, etc.). Also, when
           tallying total fish at the end  of the designated fish sampling reach, calculate the percentage of
           irruptive species to total individuals captured. If any single irruptive species comprises greater
           than 50% of the total sample, continue fishing one or more additional subreaches until the
           percentage of the irruptive species decreases to  less than 50%.
           Table 10.2 Summary of Wadeable Fishing Protocols
          Small Wadeable (less than 12.5 meters wide)
              •   Sampling reach will be between 150 and 500 meters
              •   Subreaches will be between 15 and 50 meters each
                  •S   Sample all 10 subreaches in their entirety from bank to bank starting at Transect A
                  S   Total button time will range from 500-700 seconds per subreach
                    •   You do not have to expend equal button time among the 10 subreaches— you can devote
                       more  button time to subreaches with more complex habitat.
                  •S   No minimum fish number
          Medium Wadeable (12.5 to 25 meters wide)
              •   Sampling reach will be between 500 and 1000 meters
              •   Subreaches will be between 50 and 100 meters each
              •   Minimum fishing length = 500 meters which will between 5 and 10 subreaches
              •   If needed, extend fishing length to end at a transect
                  S   Fish each subreach in a swath 8 meters from bank in pairs of subreaches starting at a
                      random bank at Transect A
                  S   Button time is roughly 700 seconds per subreach
                    •   Depending upon the habitat complexity, you can vary the distance actively fished to
                       allocate the available button time throughout the subreach.
 LJJ               •/   Minimum fish number is 500  unless all 10 subreaches have been fished.
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 <                   >  After fishing at least 500  meters, if 500 fish have not been collected, add subreaches
 CQ                      one at a time (but fish them in their entirety) until 500 fish are collected  or all 10
 ^                      subreaches have been fished.
 1/3       Large Wadeable (25 + meters wide)
 <       — - -
 S           •   Sampling reach will be between 1000 and 4000 meters
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    •   Subreaches will be between 100 and 400 meters each
    •   Minimum fishing length = 5 subreaches (which will equal between 500 and 2000 meters)
        •S  Fish each subreach in a swath 8 meters from bank in pairs of subreaches starting at a
           random bank at Transect A
        S  Button time is roughly 700 seconds per subreach
          •    Depending  upon the habitat complexity, you can vary the distance actively fished to
              allocate the available button time throughout the subreach.
        •S  Minimum fish number is 500 unless all 10 subreaches have been fished.
           >  After fishing 5 subreaches, if 500 fish have not been collected, add subreaches one at a
               time (but fish them in their entirety) until 500 fish are collected or all 10 subreaches
               have been fished.
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                     Small Wadeable Stream: Channel Width < 12.5 m
               <	 Fish Entire Reach (40 x Channel Width) —
                                                                                      K
       Medium Wadeable Stream: Channel Width  12.5 m to 25 m
                                   Fish a minimum of 500 m*, but do not
                                       stop in middle of a subreach
                                                                                       K
           Example: If average CW = 20 meters,
           each subreach equals 80 meters.
           500 meters would fall between Transects G and H.
           Initial fishing reach would stop at Transect H and equal 560 meters.
                         Large Wadeable Stream: Channel Width > 25 m
                                                    Fish a minimum of 5
                                                       subreaches*
                                                   (20 x Channel Width)
                                                                           K
 *At medium & large streams, if < 500 individuals have been collected after minimum sampling reach, continue fishing
 to next transect (alternating banks) until 500 individuals are collected or Transect K is reached, (10 subreaches fished)
Figure 10.3 Reach Layouts for Fish Sampling at Wadeable Sites

Dark shaded areas indicate the minimum length of the fish sampling reach. Light shaded areas are
sampled as needed to meet the required 500 individuals.
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10.3.2  Small Wadeable Streams
Table 10.3 describes the procedure for collecting fish in small wadeable streams. The sampling
crew should consist of one electrofisher operator, one dip-netter (1/4" mesh dip net), and an
optional bucket carrier (who may also have a net to aid in transferring fish to the livewell). For
safety, all crew members are required to wear non-breathable waders and insulated gloves. To
aid vision, wear polarized sunglasses and a hat or visor. Begin sampling at the downstream
stream end of the support reach defined for the site (Figure 10.3) and proceed upstream.
Sample the entire support reach, which will be between 150 and 500 m long. Total button time
will vary between 500 and 700 seconds per subreach. Conduct sampling by subreach  (area
between transects), but you do not have to allocate effort equally among all 10 subreaches.

Whenever possible, process fish at the end of each subreach to minimize mortality and stress to
fish. You can  use  multiple lines per species on the Fish Collection Form if necessary (e.g., you
collect a large number of individuals and need additional space for tallying, or collect  the same
species at non-adjacent subreaches [e.g., A-B and G-H]).
Table 10.3 Procedure: Electrofishing (Small Wadeable Streams)
 Electrofishina Procedures in Small Wadeable Streams
 1)  Complete the header section of the fish gear form (Site ID and date).
 2)  Decide if you will be able to sample the site.
     a)  Review all collecting permits to determine if any sampling restrictions are in effect for the site. In
         some cases, you may have to cease sampling if you encounter certain State or Federally listed
         species. If you cannot sample at all because of permit restrictions, mark Wot Fished- No Permit.
     b)  If site conditions prevent barge or backpack electrofishing (e.g., no access, safety concerns,
         conductivity is too high or too low to use a barge or backpack electrofishing unit), determine if
         you can sample by seining.
         i)   If yes, follow the procedures presented in Table 10.5.
         ii)   If not, mark Wot Fished - Site Conditions Prohibit Sampling. Note the  conditions in the
            Sampling Protocol Comments.
     c)  If you can determine that > 50% of the required fish sampling reach cannot be sampled, mark Wot
         Fished/Fishing Suspended - Can't sample >=50% of required reach.
     d)  If you cannot sample because of equipment problems, mark Not Fished-Equipment Failure.
     e)  If you cannot sample for any other reason, mark Wot Fished - Other and note the reason in the
        Explain field
 3)  If you can begin to sample, mark Wadeable in the Fish Sampling Protocol section.
     a)  Proceed to the downstream end of the support reach (Transect A)
     b)  For safety, everyone must wear personal floatation devices, non-breathable waders, foot
         protection, and insulated linesman's gloves
     c) To aid vision while netting fish, wear polarized sunglasses and a hat or visor.
 4)  Mark the appropriate Water Visibility conditions on the form. Poor implies that your ability to
     electrofish effectively is compromised because of poor visibility. Record the water temperature and
     conductivity.
 5)  Mark either Backpack, Bank or Towed Barge in the Primary Electrofishing Gear section of the fish gear
     form,
     a)   Do not use any secondary electrofishing gear in a small wadeable stream.
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                a)  Set unit to pulsed DC and mark it in the Wave Form section of the fish gear form.
                b)  Test settings outside of the sampling area. Start the electrofisher, set the timer, and depress the
                    switch to begin fishing. Typical settings are:
                    i)   500-1000 V DC
                    ii)  8-20 A
                    iii) 120 Hz.
                c)  If fishing success is poor, increase the pulse width first and then the voltage. Increase the pulse
                    rate last to minimize mortality or injury to large fish. If mortalities occur, first decrease pulse rate,
                    then voltage, then pulse width.
                d)  If no fish can be collected or electrofishing is ineffective due to high conductivity or extreme
                    turbidity, refer to seining protocol in Table 10.5.
            7)  Operation of Backpack Electrofisher—
                a)  Set unit to pulsed DC and mark it in the Wave Form section of the fish gear form.
                b)  Select the initial voltage setting based on the conductivity of the river.
                    i)   150-400 V for high conductivity (>300 u.S/cm).
                    ii)  500-800 V for medium conductivity (100 to 300 u.S/cm).
                    iii) 900-1100 V for low conductivity (< 100 u.S/cm).
                c)  Select the initial pulse rate and width.
                    i)   In waters with strong swimming fish (length >200 mm [8 in]), use a pulse rate of 30 Hz with a
                        pulse width of 2 m/sec.
                    ii)  If you expect mostly small fish, use a pulse rate of 60-70 Hz.
                d)  Turn the electrofisher on, set the timer, and depress the switch to begin fishing. If fishing success
                    is poor, increase the pulse width first and then the voltage. Increase the pulse rate last to
                    minimize mortality or injury to large fish. If mortalities occur, first decrease  pulse rate, then
                    voltage, then pulse width.
                e)  If no fish can be collected or electrofishing is ineffective due to high conductivity or extreme
                    turbidity, refer to seining protocol in Table 10.5.
                f)  Once you have  determined the appropriate settings, record them on the fish gear form. Start
                    cleared clocks and resume fishing.
                    i)   Note: some electrofishers do not meter all the requested settings; provide what you can.
                g)  If button time is not metered, estimate it with a stop watch and flag the data.
            8)  Once the settings on the electrofisher are adjusted properly to sample effectively and minimize injury
                and mortality, begin sampling at the downstream end of the reach (Transect A) and fish in an
                upstream direction.
                a)  The minimum reach length is 150 m. The maximum reach length for this protocol is 500 m.
                    i)   Search for fish even if the stream is extremely small, and it appears that sampling may
                        produce no specimens.
                    ii)  Button time should range from 500 to 700 sec per subreach. Actual time will depend upon
                        the water conditions, the diversity and complexity of the available habitat, and on the
                        number offish present in the support reach.
 LJJ
 W                     (1) If the electrofisher is highly effective and the fish are staying stunned longer and the
 ^                         water  is clear and flowing slowly then button time will be much lower than it will be in a
 ^                         system where it is more  turbid, flowing faster, and the fish are not being stunned as well.
 LJJ
 $                     (2) You do not have to expend equal button time among the 10 subreaches—you can devote
 ^                         more button time to subreaches with more complex habitat and less time to subreaches
 S£!                         with simple habitat.
 U-

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             (3) Consider that the sampling effort should provide a sample of -500 individuals, if these
                numbers exist within the support reach.
     b)   Depress the switch and slowly sweep the electrode from side to side.
     c)   Sample all habitat types (deep, shallow, fast, slow, complex, and simple). Avoid the temptation to
         focus sampling only in the richest habitat types.
         i)   For available cut-bank and snag habitats, move the anode wand into cover with the current
             off, turn the anode on when in the cover, and then remove the wand quickly to draw fish out.
         ii)   In fast, shallow water, sweep the anode and fish downstream into a net.
         iii)  In stretches with deep pools, fish the margins of the pool as much as possible, being
             extremely careful not to step or slide into deep water.
     d)   Keep the cathode near the anode if fish catch is low.
 9)  The netter, with the net 1 to 2 ft from the anode, follows the operator, nets stunned individuals, and
     places them in a bucket.
 10) Continue upstream until you reach the next transect (end of subreach).
     a)   Process fish and/or change water after each subreach to reduce mortality and stress.
         i)   Although not required, you may note amphibians and reptiles captured on the fish collection
             form.
     b)   Release fish in a location that eliminates the likelihood of recapture.
 11) Repeat Steps 8-10 until all 10 subreaches are sampled (i.e., you reach transect K).
 12) After sampling all 10 subreaches, record the final length of the fish sampling reach in the Fish Sampling
     Protocol section of the fish gear form.
     a)   If you suspend sampling before completing all 10 subreaches, record the actual length that was
         sampled, and mark the reason for the suspension in the Fish sampling - Not Conducted or
         Suspended section of the fish gear form.
     b)   If you did not collect any fish, mark Fished - None Collected in the Fish sampling - Not Conducted
         or Suspended section of the fish gear form.
 13) In the Primary Electrofishing Gear section of the fish gear form, record the total button time  expended
     for electrofishing, the total time spent sampling, and the  percentage of the total fish sampling reach
     (recorded in Step 12) sampled by electrofishing (should be 100%).

10.3.3  Medium and Large Wadeable Streams

Table 10.4 describes the procedure for collecting fish in medium and large wadeable streams.
The electrofishing crew should consist of one electrofishing operator, and one dip netter and an
optional bucket carrier (who may also have a net to aid in transferring fish to the livewell). For
safety, all crew members are required to wear non-breathable waders and insulated gloves.
Polarized sunglasses and caps to aid vision are also required.

For medium wadeable streams with a mean channel width between 12.5 and 25 m, the
minimum fish sampling reach is ~500 m (Table 10.2). If 500 m falls within a subreach, complete
sampling the subreach (so you will sample between 5 and 10 subreaches). For large wadeable
streams with a mean channel width >25 m, the minimum fish sampling reach is 20 channel
widths (5 subreaches).

As shown in Figure 10.3, begin sampling at Transect A on  a randomly determined bank and fish a
section of the subreach that extends approximately 8 m from the bank in an upstream direction.
For streams <16 m wide, or wider streams with simple habitat, fish the subreach in its entirety
(from bank to bank). Within each subreach, fish the near  bank habitat as well as midstream
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           habitat within the 8-m sampling area for a button time of ~700 seconds. When 700 seconds are
           reached, stop electrofishing unless you are "pushing" a large school offish, in which case
           continue fishing until you capture them at a break. To reduce stress and mortality, net
           immobilized fish immediately and deposit into a bucket or live-well for processing.

           Whenever possible, process fish at the end of each subreach to minimize mortality and stress to
           fish. You can use multiple lines per species on the fish collection form if necessary (e.g., you
           collect a large number of individuals and need additional space for tallying, or collect the same
           species at non-adjacent subreaches [e.g., A-B and G-H]).

           At the end of the designated fish sampling reach, determine if you have collected at least 500
           individuals. If so, stop sampling. If not, sample additional subreaches (one at a time) until at
           least 500 individuals are captured. Stop sampling when you reach Transect K (the end of the
           entire support reach), regardless of the number of individuals collected. Once the decision is
           made to fish an additional subreach, it should be completely fished as described above (do not
           stop sampling partway through a subreach).

           Table 10.4 Procedure: Electrofishing (Medium and Large Wadeable Sites)
                                  ; in Medium and Large Wadea.
            1)  Complete the header section of the fish gear form (Site ID and date).
            2)   Decide if you will be able to sample the site.
                a) Review all collecting permits to determine if any sampling restrictions are in effect for the site. In
                   some cases, you may have to cease sampling if you encounter certain State or Federally listed
                   species. If you cannot sample at all because of permit restrictions, mark Wot Fished- No Permit.
                b) If site conditions prevent barge or backpack electrofishing (e.g., no access, safety concerns,
                   conductivity is too high or too low to use a barge or backpack electrofishing unit), determine if
                   you can sample by seining.
                   i)   If yes, follow the procedures presented in Table 10.5.
                   ii)  If not, mark Wot Fished - Site Conditions Prohibit Sampling. Note the  conditions in the
                       Sampling Protocol Comments.
                c) If you can determine  that > 50% of the required fish sampling reach cannot be sampled, mark Wot
                   Fished/Fishing Suspended - Can't sample >=50% of required reach.
                d) If you cannot sample because of equipment problems, mark Not Fished-Equipment Failure.
                e)  If you cannot sample for any other reason, mark Wot Fished - Other and note the reason in the
                   Explain field
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            3)   If you can begin to sample, mark Wadeable in the Fish Sampling Protocol section.
                a)  Proceed to the downstream end of the support reach (Transect A)
                b)  For safety, everyone must wear personal floatation devices, non-breathable waders, foot
                   protection, and insulated linesman's gloves
                c)  To aid vision while netting fish, wear polarized sunglasses and a hat or visor.
            4)   Mark the appropriate Water Visibility conditions on the form. Poor implies that your ability to
                electrofish effectively is compromised because of poor visibility. Record the water temperature and
                conductivity.
5)   Mark either Backpack or Bank or Towed Barge in the Primary Electrofishing Gear section of the fish
    gear form,
    a)  Do not use any secondary electrofishing gear in a large wadeable stream.
            6)   Operation of Bank or Towed Barge Electrofisher—
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 Electrofishing Procedures in Medium and Large Wadeable Streams
     a)  Set unit to pulsed DC and mark it in the Wave Form section of the fish gear form.
     b)  Test settings outside of the sampling area. Start the electrofisher, set the timer, and depress the
         switch to begin fishing. Typical settings are:
         i)   500-1000 V DC
         ii)   8-20 A
         iii)  120 Hz.
     c)  If fishing success is poor, increase the pulse width first and then the voltage. Increase the pulse
         rate last to minimize mortality or injury to large fish. If mortalities occur, first decrease pulse rate,
         then voltage, then pulse width.
 7)
Operation of Backpack Electrofisher
a)   Set unit to pulsed DC and mark it in the Wave Form section of the fish gear form.
b)   Select the initial voltage setting based on the conductivity of the stream.
    i)   150-400 V for high conductivity (>300 u.S/cm).
    ii)   500-800 V for medium conductivity (100 to 300 u.S/cm).
    iii)  900-1100 V for low conductivity (< 100 u.S/cm).
    Select the initial pulse rate and width.
    i)   In waters with strong swimming fish (length >200 mm [8 in]), use a pulse rate of 30 Hz with a
        pulse width of 2 m/sec.
    ii)   If you expect mostly small fish, use a pulse rate of 60-70 Hz.
    Turn the electrofisher on, set the timer, and depress the switch to begin fishing. If fishing success
    is poor,  increase the pulse width first and then the voltage. Increase the pulse rate last to
    minimize mortality or injury to large fish. If mortalities occur, first decrease pulse rate, then
    voltage, then pulse width.
    If no fish can be collected or electrofishing is ineffective due to high conductivity or extreme
    turbidity, refer to seining protocol  in Table 10.5.
    Once you have determined the appropriate settings, record them on the fish gear form. Start
    cleared  clocks and resume fishing.
    i)   Note: some electrofishers do not meter all the requested settings; provide what you can.
    If button time is not metered, estimate it with a stop watch and flag the data.
     c)



     d)


     e)

     f)
     g)
 8)  Once the settings on the electrofisher are adjusted properly to sample effectively and minimize injury
     and mortality, begin sampling at the downstream end of the reach (Transect A). Randomly choose a
     bank on which to start and fish in an upstream direction within 8 m of the shoreline.
     a)  For the medium stream protocol, the minimum initial fish sampling reach length is 500 m. If 500
         m or 2000 m falls within a subreach, finish sampling the subreach.
     b)  For the large stream protocol, the minimum initial fish sampling reach length is 5 subreaches
         i)   For streams <16 m wide, or wider streams with simple habitat, fish the subreach in its
             entirety (from bank to bank).
 9)  When using a towed electrofishing unit (barge or pram), the minimum crew size for electrofishing is
     three.
     a)  The barge operator must remain actively at the control box and navigate the barge.
     b)  The probe operator will use one probe.
 10) When using a backpack electrofishing unit, the minimum crew size is two (one operator with the
     probe and one netter).
 11) Depress the switch and slowly sweep the electrode from side to side.
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            Electrofishing Procedures in Medium and Large Wadeable Streams
                a)   Search for fish even if it appears that sampling may produce no specimens.
                b)   Sample all habitat types (deep, shallow, fast, slow, complex, and simple). Avoid the temptation to
                    focus sampling only in the richest habitat types.
                    i)   For available cut bank and snag habitats, move the anode wand into cover with the current
                        off, turn the anode on when in the cover, and then remove the wand quickly to draw fish out.
                        In fast, shallow water, sweep the anode and fish downstream into a net.
                        In stretches with deep pools, fish the margins of the pool as much as possible, being
    c)
                        extremely careful not to step or slide into deep water.
                    Keep the cathode near the anode if fish catch is low.
            12) The netter, with the net 1 to 2 ft from the anode, follows the operator, nets stunned individuals, and
                places them in a bucket.
            13) Continue upstream until you reach the next transect (end of subreach).
                a)   The total button time within each subreach should be -700 sec. depending upon the habitat
                    complexity, you can vary the distance actively fished to allocate the available button time
                    throughout the subreach.
            b)      Process fish after each subreach to reduce mortality and stress. Release fish in a location that
                    eliminates the likelihood of recapture.
                    i)   Although not required, you may note amphibians and reptiles captured on the fish collection
                        form.
            14) Move to the opposite bank when necessary (see Figure 10.3). Repeat Stepsl2-14 until you have
                sampled the required length of stream (Step 8).
                a)   If you have to suspend sampling before completing the required fish sampling reach, record the
                    actual length that was sampled, and mark the reason for the suspension in the Fish sampling -
                    Not Conducted or Suspended section of the fish gear form.
            15) If the required length did not include all 10 subreaches, determine the total number of individuals
                collected.
                 a)   If the total is < 500, sample one or more additional subreaches until at least 500 individuals have
                     been collected and processed, or you sample all 10 subreaches. Go to Step 17.
                 b)   If you collect > 500 individuals, determine if a single irruptive species comprises > 50% of the
                     total  number of individuals.
                       i) If an irruptive species make up > 50% of the sample, sample one or more additional
                        subreaches to bring the proportion of the irruptive species below 50%. Go to Step 17.
                       ii) If not, go to Step 17.
                c)  If you  have sampled all 10 subreaches (i.e., you have reached Transect K), go to Step 17.
            16) After sampling, record the final length of the fish sampling reach in the Fish Sampling Protocol section
                of the fish gear form.
                a)  If you did not collect any fish, mark Fished - None Collected in the Fish sampling - Not Conducted
                    or Suspended section of the fish gear form.
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17)  In the Primary Electrofishing Gear section of the fish gear form, record the total button time expended
    for electrofishing (should be -700 sec per subreach sampled), the total time spent sampling, and the
    percentage of the total fish sampling reach (recorded in Step 15 or 17) sampled by electrofishing
    (should be 100%).
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10.4   Seining
In small or large wadeable streams where conditions prohibit electrofishing, use seining only as
the last option for collecting fish. If conditions are such that seining is the only method used,
provide a justification in the Sampling Protocol Comments section of the fish gear form (Figure
10.1). Table 10.5 presents the procedure for seining wadeable streams. The intent of the seining
effort is to provide comparable data to electrofishing.
Although electrofishing typically works best in an upstream direction, seining may work best
moving downstream. Allocate seine hauls so that the snag, edge, and mid-channel habitats are
fished thoroughly. In general, edge and snag habitats will be sampled using narrower seines
over shorter distances, while mid-channel habitats will be sampled using longer seines over
longer distances. Generalized habitat seining procedures are presented in Table 10.5.
Depending upon habitat types and complexity, use 2 to 3 crew members. Two crewmembers
move the seine; an optional third person creates and maintains a bag in the seine in area with
higher velocities, or agitates rocks in riffles or snags. To avoid mortality, process fish after each
seine haul. If necessary, you can use additional lines on the fish collection form (Figure 10.2) to
record species collected with different nets or in different hauls.
If you seine, record information for each seine haul on the Seining Information Form to track
effort ( Figure 10.4).  Denote the bank as right or left as you face downstream. Restrict each haul
to a single habitat type. After fish sampling is completed for the site, use the information from
the seining information form to complete the information in the Primary Seine Net section of
the fish gear form (Figure 10.1). Include the seining information form with the packet of
completed field forms you submit to the NRSA information management staff.
Table 10.5 Procedure: Seining (Wadeable Sites)
 Procedures for Seining at.
/adeable Sites
 1)  Use seining as a last option only (e.g., when electrofishing is ineffective due to high conductivity or
     extremely high turbidity). Do not use seining as a supplementary method to electrofishing. Fish
     sampling reach must be safely wadeable.
     a)   If site conditions are such that only seining is used, note the reason in the Sampling Protocol
         Comments section.
     b)   At the end of each seine haul, immediately place all fish in one or more live wells to minimize
         injury and mortality, and so that most fish can be returned to the river alive.
 2)  Complete the header section of the fish gear form (Site ID and date).
 3)  Mark the pertinent protocol and size class in the Fish Sampling Protocol section.
     a)   Proceed to the downstream end of the support reach (Transect A).
         i)   At some sites, seining may be more effective while working downstream (from Transect K)
             instead of upstream.
             (1)  If working downstream in a large wadeable stream, reverse the transects in Figure 10.3
                and move to the opposite bank where indicated.
     b)   For safety, everyone must wear personal floatation devices and foot protection.
     c)   To aid vision while seining, wear polarized sunglasses and a hat or visor.
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 4)  Mark the appropriate Water Visibility conditions on the form. Poor implies that your ability to seine
     effectively is compromised because of poor visibility. Record the water temperature and conductivity.
 5)  Mark the type of seine being used (Bag Seine or Minnow Seine) in the Primary Seine Net section of the
     fish gear form. This is the seine that will be used for sampling the majority of the fish sampling reach.
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                a)  Record the number of crewmembers (2-3), and the net dimensions (height, total length, and
                    mesh size) on the fish gear form.
                b)  If you have to use a second type of seine for parts of the sampling reach, Mark the type and
                    record the dimensions in the Secondary Seine Net section of the fish gear form.
            6)  Determine the length of the fish  sampling reach and the number of subreaches that should be
                sampled (refer to Table 10.2).
            7)  To maximize capture efficiency, please do the following:
                a)  Always use 10 and 20 ft. seines. When necessary, reduce the width by rolling seine poles and
                    floats into the net.
                b)  When narrowing seines, always keep lead line outside of the pole.
                c)  When working edge habitats, only roll the inner side of the seine, while keeping the near bank
                    pole extended.
                d)  As a default, use seines that are 2 meters in depth. A 1.25 meter seine may be  used in shallow
                    habitats.
                e)  Keep the float line above the surface (avoid dragging it below the surface while pulling).
                f)  Maintain the lead line along the river bottom.
                g)  Either tie the seine to the poles tightly, or roll the seine into the poles.
                h)  Always maintain the bag behind the poles.
            8)  Seining habitats include large riffles or gravel bars, pools (which include backwater  areas), glides or
                runs, edges, and snags. Seine width and haul length is dependent upon the water velocity, depth,
                and/or complexity of the habitat.
                a)  The objective of the seining  effort is to acquire a comparable collection of fish  (in terms of species
                    richness and relative abundance, and allocation of effort throughout the fish sampling reach) to
                    that obtained if the site was electrofished.
                    i)   Avoid extended seine hauls that collect hundreds of individuals.
                    ii)   Seine as many available habitat types as  possible within each subreach (one haul each).
                    iii)  Total time spent seining a site should be  comparable to what would have been spent
                        electrofishing.
                b)  Riffle Habitats
                    i)   Use two crewmembers, each tending a seine pole. Place the seine perpendicular to the
                        current across the downstream end of the riffle. Ensure that the lead  line is on the bottom.
                        Tilt the net slightly downstream to form a bag  to trap aquatic  vertebrates.
                    ii)   Starting no more than 3 m upstream, a third crewmember kicks the substrate and overturns
                        rocks, proceeding quickly downstream toward the net.
                    iii)  When the area is thoroughly kicked, quickly raise and bag the net. .  Process fish (i.e.,
                        enumerate, identify and voucher fish) and record tally information on the  fish collection form
                        (Figure 10.2). You may use separate lines on the fish collection form to record species
                        information from seine  hauls.
                    iv)  Record seine characteristics and haul length, habitat, and time on the seining information
                        form (  Figure 10.4).
 LJJ             c)  Pool, Backwater, and Bar Habitats (Slack water)
 (D
 fj                 i)   Use two crewmembers, each tending a seine pole. Pull the seine across the pool using
 S                     shallow riffles or banks as barriers. A third crewmember creates and maintains a bag in the
 £J                     seine.
 to
 <                 ii)   In areas with current, pull the net downstream and then sweep toward the bank with one or
 ^                     both poles, or post one  pole on the bank and sweep the other end in a wide arc from
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             midstream to the same bank.
         iii)  You can work pools in short to long hauls and use seines of varying width depending on the
             complexity and depth of the pool. Keep the seine depth constant at 2 meters.
         iv)  Pull the bag completely to shore at a predesignated point.
     d)   Glide or Run Habitats (noticeable current)
         i)   Use two crewmembers, each tending a seine pole. Pull the seine diagonally across the glide
             towards the bank. If necessary, a third crewmember creates and maintains a bag in the seine.
         ii)   Pull the net quickly downstream along the glide moving diagonally toward  the bank. When
             you reach the bank with the outer edge of the seine, post the pole and  sweep the other end
             in a wide arc from midstream to the same bank.
         iii)  Because of decreased complexity and shallower depths, seine hauls in glides or runs are
             typically longer and use wider nets. You can use a 1.25 m deep seine in  shallow glides.
         iv)  Pull the bag completely to shore at a predesignated point.
     e)   Edge Habitats
         i)   Edge habitats may be shallow too deep with complex to uniform habitat, and may include
             undercut banks.
             Use two crewmembers, each tending a seine pole. Seine along the nearshore area.
             The near bank crewmember moves along the shore while jabbing along any undercut or small
             structure. The other crewmember stays ahead of the shoreline pole to  maintain a "J" in the
             seine bag. At a predesignated point, post the near shore pole and sweep the seine towards
             and up on the bank.
         iv)  Depending on edge complexity and depth, seine width and haul length  may vary. Use wider
             seines and longer hauls in shallower, less complex habitats. As complexity, depths, and flow
             increase, shorten the seine width and haul length accordingly. Seine depth may vary
             depending on depth.
     f)   Snag Habitats
         i)   Snag habitats often require  creativity in terms of seine length and approach. You can use  a
             1.25 meter deep seine to avoid snagging the net on structure, but use a 2 m deep seine in
             deeper areas. Narrow seine widths and short hauls are preferred.
         ii)   Use two crewmembers, each tending a seine pole. Jab seining is often the most effective
             method. Quickly jab a shortened seine (< 2 m wide) under the cover and near the river
             bottom, then quickly lift the seine to the water surface. You can use a third crewmember  to
             agitate the snag to move fish out toward the seine.
         iii)  For small snags along the bank, seining along the edge may work best. The near snag
             crewmember moves along the snag, while jabbing along its length. The other crewmember
             stays ahead of the shoreline pole to maintain a "J" in the seine bag. At a predesignated point,
             quickly pull the seine to the surface.
 9)  To minimize mortality, process fish (i.e., identify, count, and prepare preserved voucher specimens or
     photovoucher images) after each seine haul (rather than at the end of a subreach.
     a)   Record identifications, tallies, and voucher information on the fish collection form (Figure 10.2).
         You may use separate lines on the fish collection form to record species information from
         separate seine hauls.                                                                            LU
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     b)   Tally seine characteristics and haul length, habitat, and time on the seining information form (           ^
         Figure 10.4).                                                                                   |
 10) At the end of all sampling, use the seining information form to  determine the total number of hauls,          1/1
     the average haul length, the total time spent seining, and the proportion of the total fish sampling            <
     reach sampled  (recorded in the Fish Sampling Protocol section  of the fish gear form) for each type  of          S
                                                                                                       LJ_


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                seine. Record the totals in the Primary and Secondary Seine Net sections of the fish gear form.
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FISH ASSEMBLAGE

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          National Rivers and Streams Assessment 2013/14                          Field Operations Manual
          Version 1.0, May 2013                                                            Wadeable

          10.5  Processing Fish
          Process fish at the end of each subreach, as described in Table 10.6. However, if fish show signs
          of stress (e.g., loss of righting response, gaping, gulping air, excessive mucus) in the middle of a
          subreach, change the water in the live well or stop fishing and initiate processing. Always
          process and release individuals of State or Federally listed threatened or endangered species or
          large game fish immediately after collection. After processing fish, release them in a location
          that prevents the likelihood of their recapture.
          If you use a seine to collect fish, please record the information on a separate line on the field
          form.

          10.5.1 Identification and Tallying
          Record species identifications, tallies, and other information for individuals collected on the Fish
          Collection Form (Figure 10.2). Use multiple pages of the form as needed to record all species
          collected. It is important to record page numbers correctly because page number is one of the
          variables used to uniquely identify a species record. You can record separate collections of the
          same species on multiple lines of the collection form (e.g., when you  encounter a species in non-
          adjacent subreaches, or collect a species with a secondary gear type). Do not process individuals
          with total length < 25 mm (1 inch), as these are likely young of year individuals that cannot be
          identified confidently to species. Only crew members designated as "taxonomic specialists" by
          EPA regional coordinators can identify fish species. Tally fish by species and major size class (6
          inch [15 cm]  intervals), and examine them for the presence of DELT (Deformities, Eroded Fins,
          Lesions and Tumors) anomalies. Use common names of species established by the American
          Fisheries Society Common and Scientific Names of Fishes from the United States, Canada and
          Mexico (Nelson, et al. 2004). Appendix D provides a list of species names reported from the
          NRSA 2008/09.
          If you believe a specimen is nonindigenous to the site, mark it as Introduced on the collection
          form. If you suspect it represents a potential range extension for the  species, prepare one or
          more specimens (preserved if possible but photographs if not). Physical specimens are  required
          in order to publish reports of range extensions. Include specimens to document suspected range
          extensions are included as part of the preserved Unknown/Range Extension voucher sample
          (UNK/RNG; Section 10.5.2).

          10.5.2 Unknown Specimens
          If you cannot positively identify individuals to species in the field, record taxonomic information
          of the collection form using scientific names rather than common names. If you can identify a
          specimen only to family, record the scientific rather than the  common family name (e.g.,
          UNKNOWN PERCID A, not UNKNOWN PERCH A) on the fish collection form. If you can identify a
          specimen to genus, record the scientific name rather than the common name (e.g., UNKNOWN
          PERCINA A, not UNKNOWN DARTER) on the fish collection form. Using scientific rather  than
          common names for unknowns reduces ambiguity, since some common names may in fact refer
 £j       to multiple genera (e.g., "darter", "shiner", "sucker", "sunfish", etc.).  If you identify an unknown
 fj       species to Genus, retain a small number (up to 20 individuals per putative species) as part of the
 pn
 ^       preserved UNK/RNG voucher sample (see Section 0) or take good  digital photographs
 to       (SectionlO.5.3) for laboratory identification. If you are only able to identify an unknown to
 **       Family, retain as many of the individuals as possible for later identification. Use the UNK/RNG
122

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             Wadeable
Voucher label on the label sheet to label your jar of unknown to track from which sites the
unknowns originated.

Table 10.6 Procedure: Processing Fish (Wadeable Sites)
 Fish Processing
 1)   Complete all header information accurately and completely on the fish collection form. It is important
      to paginate the collection forms correctly (e.g., start with page 1, do not duplicate page numbers,
      etc.), as page number is part of the unique record identifier for the fish count data.
 2)   Process individuals collected at the end of each subreach. You can record a single species on multiple
      lines of the collection form (e.g., use separate lines for individuals collected in nonadjacent
      subreaches, collect with a secondary gear type, or if you need additional space to record tally marks,
      etc.).
      a)   Process species listed as threatened and endangered first as described in Step 4.
            i)     Photograph specimens for voucher purposes if conditions permit and stress to individuals
                 will be minimal. Mark as Photo in the Voucher section of the collection form.
            ii)    If individuals die due to sampling, prepare them as part of the local voucher sample and
                 preserve them in the field. Comply with the conditions of your collection permit in
                 regards to mortality of listed species.
            iii)   Return individuals to the river immediately after processing.
 3)   Only identify and process individuals > 25mm (1 inch) in total length (TL). Ideally handle specimens
      only once.
 4)   Identify each individual to the lowest possible taxonomic level:
      a)   If you can confidently identify the individual to species, record the common name on the first
          blank line in the Common Name field of the fish collection form.
            i)    Common names should follow those recognized by the American Fisheries Society. Use
                 of alternative names is discouraged. Use names presented in Appendix D, which are
                 based on those used in the NRSA 2008/09.
              (1)  Record the complete common name. Avoid using shortened names (e.g., stoneroller,
                  carp, bass, etc.).
              (2)  If you use a non-standard name, you must assign a flag to the line and provide the
                  taxonomic reference for the name in the Comments section of the collection form.
      b)   If you cannot positively identify an individual to the species level:
            i)     Identify it to the lowest taxonomic level (i.e., family or genus). Record the putative
                  name as UNKNOWN plus the scientific name of the family or genus (e.g., UNKNOWN
                  CATOSTOMID A, UNKNOWN MOXOSTOMA A) in the Common Name column of the
                  collection form.
            ii)     If you are permitted to retain the specimen, assign it the next available sequential tag
                  number (starting with 01) in the Voucher Tag Number column and see Step 9.
      c)   If you believe the individual is a hybrid:
                  Mark as Hybrid? on the collection form.
                  If the hybrid has an accepted standard common name (e.g., Tiger muskellunge,
                  Saugeye, Wiper, etc), record that name. For other hybrids record the common name of
                  both species (e.g., Green sunfish x Bluegill, Cutthroat trout x Rainbow trout). Avoid using
                  non-specific terms such as Hybrid sunfish.
              \    If you are unsure of the identification and are permitted to retain the specimen, assign
              it the next available sequential tag number (starting with 01) in the Voucher Tag Number
              column and see Step 9.
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            5)    If you know the species is not native to this location, mark as Introduced?
            6)    Visually estimate the total length of each individual (a measuring board is not necessary). Keep a
                 running tally in the appropriate Tally and Counts section (< 6 in., 6-12 in., 12-18 in., or > 18 in.) of the
                 fish collection form.
                 a)   If all individuals of a species appear to be the same size, provide a flag and comment for the line
                     if you believe the population is stunted.
            7)    Examine each individual for external anomalies. Readily identify external anomalies including missing
                 organs (eye, fin), skeletal deformities, shortened operculum, eroded fins, irregular fin rays or scales,
                 tumors, lesions, ulcerous sores, blisters, cysts, blackening, white spots, bleeding or reddening,
                 excessive mucus, and fungus. After you process all of the individuals of a species, record the total
                 number observed in tbeAnom Count column of the collection form.
            8)    If an individual has died due to electrofishing or handling, include it in the running tally for the
                 species. After you process all of the individuals of a species, record the total number observed in the
                 Mortality/Count column of the collection form.
            9)    If you are retaining individuals of the species as part of the preserved Unknown/Range Extension
                 Unk/Rng) voucher sample:
                 a)   Mark as Unk/Rng in the Voucher section of the collection form.
                 b)   Assign the species the next available voucher specimen tag, and record the number in the
                     Voucher Tag # column of the collection form.
                      i)       If you take one or more photographs of the species instead of preserving specimens,
                              assign the next available voucher specimen tag number in the Voucher Tag # column of
                              the collection form. Include the specimen tag in all photos of the species. Mark Photo
                              in the  Voucher section of the collection form.
                       (1)    Ideally, take photos of all species collected at a site that are not being preserved.
                 c)   Record the number of individuals retained for the preserved voucher sample in the Vouchers
                     Retained column of the collection form.
                      i)       NOTE: Do not keep separate tallies of voucher and non-voucher specimens. Record all
                              individuals in the appropriate area of the Tally and Counts section. The retained
                              voucher specimens represent a subsample of the total count.
                      ii)     Place the specimens in a jar which has been labeled with the site ID. You can have
                             multiple individuals of the same species in the jar, but each species will have a separate
                             voucher tag number (i.e. one tag number per line on the collection form).
            10)  If you are retaining specimens as part of a preserved QA voucher sample for the site:
                 a)   Mark as QA in the Voucher section of the fish collection form.
                      i)      NOTE: This should be marked at least once for all species collected at the site (including
                             unknowns).
                 b)   Use the sheet of labels and tags for the QA voucher sample (the jar label has a preprinted sample
                     ID number). Assign the species the next available voucher specimen tag number. Record the
                     specimen tag number in the Voucher Tag # column of the collection form.
                      i)      If you take one or more photographs of the species instead of preserving specimens,
                             assign the next available voucher specimen  tag number, and record the number in the
                             Voucher Tag # column of the collection form. Include the specimen tag in all photos of
                             the species. Mark Photo in the Voucher section of the collection form.
                 c)   Record the number of individuals retained for the preserved voucher sample in the Vouchers
                     Retained column of the collection form.
                      i)      NOTE: Do not keep separate tallies of voucher and non-voucher specimens. Record all
                             individuals in the appropriate area of the Tally and Counts section. The retained voucher
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                 specimens represent a subsample of the total count.
           ii)    Place the specimens in a fine mesh bag (or separate jar) along with the voucher
                 specimen tag that matches the number recorded on the collection form. You can have
                 multiple bags (or jars) of the same species, but each bag (or jar) will have a separate
                 voucher tag number (i.e., one tag per line on the collection form).
 11)   Repeat Steps 2 through 10 for each subreach sampled. Use additional fish collection form sheets as
      needed, being careful to paginate each sheet correctly.
 12)   Record the fish collected with seining methods on a separate line on the field form.
 13)   At the end of sampling, follow the appropriate procedure to prepare the preserved voucher samples
      (UNK/RNG and/or QA) and/or select specimens for tissue samples.
      a)   For all voucher samples, use a sufficient volume of 10% buffered formalin (the volume of
          formalin solution used must exceed the volume of specimens). Use additional jars if necessary.
          Slit large individuals (TL > 200 mm  [~8 in.]) along the right side in the lower abdominal cavity to
          allow penetration of the formalin solution.
 14)   Complete a sample jar label for the UNK/RNG voucher sample. Attach it to the sample jar and cover it
      with clear tape.
 15)   If you did not prepare a QA voucher sample, mark Wo Voucher Preserved on the back of the fish gear
      form.
      a)   Otherwise complete a sample jar label for the QA voucher sample. Attach it to the sample jar and
          cover it with clear tape.
      b)   Record QA voucher sample label information on the back of the fish gear form.
 16)   Record the file names of any photovouchers taken on the back of the fish gear form.
      a)   Use only one line per voucher tag, even if you took multiple photos (record the beginning and
          end of the sequence in the Sequence column). Make sure the page and line numbers you record
          match those on the collection form.
      b)   Name image files as: Site ID + Visit number + tag number + sequence (e.g.,
          CTLS1001_Vl_tag01a).
 17)   If you did not collect any fish from the entire fish sampling reach, mark Fished - None Collected in the
      Fish Sampling - Not Conducted or Suspended section of the fish gear form.
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          10.5.3 Photovouchering
          Use digital imagery for fish species that cannot be preserved as voucher specimens (e.g., rare,
          threatened, and endangered species; very large bodied). Ideally, take photos of all species
          collected a site (that are not preserved) to provide a minimal level of documentation of
          occurrence. Take photographs of entire specimens and additional specific morphological
          features that are appropriate  and necessary for an independent taxonomist to accurately
          identify the specimen. Additional detail for these guidelines is provided in Stauffer et al. (2001),
          which is provided to all field crews as a handout.
          The recommended specifications for digital images to be used for photovouchering include: 16
          bit color at a minimum resolution of 1024x768 pixels; macro lens capability allowing for images
          to be recorded at a distance of less than 4 cm; and built-in or external flash for use in low-light
          conditions. Specimens (or morphological features) should occupy as much of the field of view as
          possible.  Use a fish measuring board, ruler, or some other calibrated device to provide  a
          reference to scale. Provide an adequate background color for photographs (e.g., fish measuring
          board). Include a card with site ID number, site name, and date in each photograph so that
          photos can be identified if file names become corrupted. In addition, include the voucher
          specimen tag that  you assign to the species to provide a link to the line on the fish collection
          form. For each photovoucher specimen, include at least a full-body photo (preferably of the left
          side of the fish), and other macro images of important morphological features (e.g., lateral line,
          ocular/oral orientation, fin rays, gill arches, mouth structures, etc.). It may also be necessary to
          photograph males, females, or juveniles.
          Save images in medium-to  high-quality jpeg format. It is important that time and date stamps
          are accurate, as this  information can also be useful in tracking the origin of photographs.
          Transfer images stored in the  camera to a personal computer (PC) or external storage device
          (e.g., thumb drive  or flash memory card) at the first available opportunity. At this time,  rename
          the original files to include the site ID, visit number, voucher specimen tag number, and photo
          sequence (e.g., CTLS1001_Vl_tag01a.jpg). Record the file names on the back of the fish gear
          form (Figure 10.6).
          Maintain a complete set of your photovoucher files in a safe location (e.g., an office computer
          that is backed up regularly) for the duration of the sampling season. At this time, you will
          transfer all images to the NRSA information management (IM) staff. You will also send copies of
          your image files as part of your QA voucher samples (Section 10.5.3).

          10.5.4 Preparing Preserved Voucher Specimen Samples
          There are two different types of samples for preserved voucher specimens. The UNK/RNG
          voucher samples are used to identify specimens that cannot be confidently identified in the
          field, and to provide physical specimens of suspected range extensions. After submitting the fish
          collection form to the NRSA IM staff, you will receive an update form that lists only the  records
          for unknown species recorded on the fish collection form (including phototvouchers) that were
 LU       marked as being part of the UNK/RNG voucher sample.
 (D
 5i       In addition to a UNK/RNG voucher sample (if needed),  you will prepare an additional  QA voucher
 ^       sample (Section 10.5.6). A QA voucher sample will be performed at a pre-designated  set of sites
 to       and includes preserved specimens (or photographs)  of all species collected at a site (including
 <       the unknowns). Use the voucher specimen tags and sample labels designated for QA voucher
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samples. QA voucher samples are eventually sent to an independent taxonomist as a check on
the accuracy of each fish taxonomist.

10.5.5 Preserving Voucher Specimen Samples
Preserve UNK/RNG and QA voucher specimens in the field with a 10% buffered formalin
solution. The volume of formalin must be equal to or greater than the total volume of
specimens. Use additional jars if necessary to ensure proper preservation. For individuals having
a total length  larger than 200 mm (~8 in.), make a slit along the right side of the fish in the lower
abdominal cavity to allow penetration of the preservative solution. Follow all the precautions for
handling formalin outlined in the MSDS. Formalin is a potential carcinogen. Handle with
extreme caution, as vapors and solution are highly caustic and may cause severe irritation on
contact with skin, eyes, or mucus membranes. Wear vinyl or nitrile gloves and safety glasses,
and always work in a well-ventilated area.
Once you have completed preserving all jars of voucher specimens, complete the appropriate
jar label (Figure 10.5 for UNK/RNG samples, and Figure 10.6 for QA voucher samples). Attach
the completed label to the jar and cover with clear shipping tape. If you have > 2 jars of either
type of sample, prepare a hand-made label for each additional jar (this is more important for the
QA voucher sample,  which has a unique sample ID number).
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          National Rivers and Streams Assessment 2013/14

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Field Operations Manual

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FISH VOUCHER - UNK/RNG
Site ID:
/ /201

FISH - BAG
TAG: 41
FtSH - BAG
TAG: 37
FtSH - BAG
TAG; 33
RSH -BAG
TAG: 29
FISH - BAG
TAG: 25
RSH - BAG
TAG: 21
FtSH - BAG
TAG: 17
FtSH - BAG
TAG: 13
FISH - BAG
TAG: 09
FISH - BAG
TAG: 05
RSH - BAG
TAG: 01
FISH - BAG
TAG: 42
FISH - BAG
TAG: 38
FISH - BAG
TAG: 34
FISH - BAG
TAG: 30
FISH - BAG
TAG: 26
FISH - (?AG
TAG. 22
FISH - BAG
TAG: 18
FISH - BAG
TAG: 14
FISH - BAG
TAG: 10
FISH - BAG
TAG: 06
FISH - BAG
TAG: 02
FISH VOUCHER - UNK/RNG
Site ID:
i 1201

FISH - BAG
TAG: 43
RSH - BAG
TAG: 39
' 	 1
FISH - BAG
TAG: 35
FISH- fWj
TAG. 31
FISH - BAG
TAG: 27
RSH - BAG
TAG: 23
RSH - BAG
TAG: 19
RSH - BAG
TAG: 15
FISH - BAG
TAG: 11
FISH - BAG
TAG: 07
FtSH - BAG
TAG: 03
FISH - BAG
TAG: 44
FISH - BAG
TAG: 40
FISH - BAG
TAG; 36
FISH - BAG
TAG: 32
FISH - BAG
TAG: 28
FISH - BAG
TAG: 24
FISH - BAG
TAG: 20
FISH - BAG
TAG: 16
FISH - BAG
TAG: 12
FISH - BAG
TAG: 08
FISH - BAG
TAG: 04
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          Figure 10.5 Unknown/Range Extension Voucher Sample Labels and Voucher Specimen Tags
128

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                                    NRSA 2013/14 FISH GEAR AND SAMPLING INFORMATION (Back)


                                                                                                  Date:        /
                                   QA VOUCHER SAMPLE INFORMATION (VERT)
                                                                                                                 NO VOUCHERS PRESERVED Q
                                    Sample 10
                                                       Oof Jars  Preserved
                                                                 O
                                                                                        Comments
                                   PHOTO VOUCHER FILE INFORMATION
          Photo File Name (SilelD_V(Vtsit#)_Tag*)   Sequence

Page  Line       (e.g.: SSCT001_V1_Tag01)        (a.g. a-c)
                                                                      Page
Photo File Name |SltelD_Vp/]slt*)_Tag*|

    (e.g.: SSCTQ01_V1_Tag01)
Sequence

 (e.g. a-c)
                                   FISH TISSUE PLUG SAMPLES (FPLG)
                                                                                                                    NO SAMPLE COLLECTED Q
                                   Collection Method:  Q ELECTROFISHING  Q HOOKS. LINE   O SEINING  Q OTHER:
                                          Sample ID
                                                                         Common Name
                                                                                                      LwiglrXmm)
                                                                                                                 Weighing)
                                            04/11/2013 2013 Fish Gear
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M      FISH ASSEMBLAGE

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          National Rivers and Streams Assessment 2013/14                          Field Operations Manual
          Version 1.0, May 2013                                                           Wadeable

          10.5.6 Processing Unknown/Range Extension (UNK/RNG) Voucher Samples
          Table 10.7 outlines the procedure for determining the identification of unknown specimens
          from each UNK/RNG sample. A more detailed procedure for conducting the laboratory
          identifications is presented in the NRSA laboratory operations manual (USEPA 2013b). Identify
          unknown specimens using whatever resources are necessary (magnification, literature,
          reference collections/specimens, including dissected anatomical features or in-house
          colleagues).
          Following positive laboratory identification, use the update form for the sample (Figure 8.10) to
          reconcile the unknown records to reflect the actual species identifications and  numbers. It is
          important to update counts and identifications by voucher tag—do not combine multiple
          samples of the same unknown before updating.
          If all specimens for an  unknown record are a single species, simply record the final identification
          (as common name from the standard list [Appendix D]) in the Revised Name column, and enter
          the original count in the Revised Count column. If you determine that a single unknown record is
          actually >1 species, you will record the new species on blank lines. For example, if a sample of
          20 specimens of UNKNOWN COTTUSA is later identified as 15 individuals of one species and 5
          individuals of another, correct the total count for the unknown assigning 75% of the original
          total count to the first species  and 25% of the original total count to the second species. Record
          the information (revised common name and revised count) for the first species on the same line
          as the original unknown record. Record the information for additional species from this original
          unknown as new data  records, but retain the page, line number, and voucher specimen tag
          number of the original unknown record. Record the name and count in the Revised Name and
          Revised Count columns.
          If you use a non-standard name, enter the page, line number, tag number, and taxonomic
          reference for the name in the Comments section of the update form. Submit your completed
          update forms to the NRSA IM staff as soon as possible after completing the laboratory
          identifications. Retain the preserved UNK/RNG voucher samples from each site. Contact your
          regional  EPA coordinator if you cannot store the samples at your facility.
          If your attempts at identification do not yield a positive identification for 100% of the fish you
          retained, contact the Field Logistics Coordinator for further guidance (Chris Turner,
          cturner@glec.com. 715-829-3737). There are provisions under which fish can be identified by a
          contracted lab and the results  returned to you.

          10.5.6 Processing QA Voucher Samples
          Prepare the QA voucher sample as outlined in Table 10.8. Prepare the QA voucher sample
          separately from the UNK/RNG voucher sample. Processing involves ensuring that the sample
          jar(s) and photovoucher files include specimens of ALL species (including unknowns and
          common species collected from the site. Each unique species (including unknowns) should have
          a unique QA voucher specimen tag assigned (Figure 10.7). Record information  about the
 LJJ       preserved QA voucher sample on the back side of the fish gear form (Figure 10.6).
 (D
 ->       Retain all of your QA voucher samples (including digital image files) until given  direction by EPA
 ^       regarding where to send them. When you are ready to ship the samples, complete a sample
 I i i
 $       tracking form as described in Appendix C. QA voucher samples may require shipping as
 **       "dangerous goods," and packing and documentation requirements will differ depending on
 S2       whether the samples contain formalin or ethanol, and on the particular shipping service used.


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QA FISH VOUCHER (VERT)
Site ID:
	 / 	 /201_
990011
FISH - BAG
TAG
41
990011
FISH - BAG
TAG
37
990011
FISH - BAG
TAG:
33
990011
FISH -BAG
TAG
29
990011
FISH -BAG
TAG
25
990011
FISH - BAG
TAG
21
990011
FISH - BAG
TAG
17
990011 s)
FISH - BAG
1*0
13
990011
FISH - BAG
TAG
09
990011
FISH - BAG
TAG
05
990011
FISH -BAG
TAG:
01
990011
FISH - BAG
TAG
42
990011
FISH -BAG
TAG
38
99001 1
FISH - BAG
TAG
34
990011
FISH-BAG
TAG
30
990011
FISH - BAG
TAG
26
99001 1
FISH - BAC
TAG
22
9S.OC11
F1SH - BAG
TAG
18
990011
FISH - BAG
TAG
14
990011
FISH - BAG
TAG
10
99001 1
FISH -BAG
TAG
06
99001 1
FISH -BAG
TAG
02
990011
QA FISH VOUCHER (VERT)
Site ID:
	 / 	 /201_
990011
FISH - BAG
TAG
43
990011
FISH - BAG
TAG
39
99001 1
FISH - BAG
?5°
99001 1
FISH - BAG
TAG
31
c\990011
rlSH - BAG
TAG:
27
990011
FISH - BAG
TAG
23
990011
FISH - BAG
TAG
19
990011
FISH - BAG
TAG
15
990011
FISH - BAG
TAG
11
990011
FISH - BAG
TAG
07
990011
FISH - BAG
TAG:
03
990011
FISH -BAG
TAG
44
990011
FISH -BAG
TAG
40
990011
FISH -BAG
TAG:
36
990011
FISH -BAG
TAG
32
990011
FISH -BAG
TAG
28
990011
FISH -BAG
TAG
24
990011
FISH -BAG
TAG
20
990011
FISH -BAG
TAG
16
990011
FISH -BAG
TAG
12
990011
FISH -BAG
TAG
08
990011
FISH - BAG
TAG
04
990011
Figure 10.7 QA Voucher Sample Labels and Voucher Specimen Tags
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           Table 10.7 Procedure: Processing Unknown/Range Extension (UNK/RNG) Voucher Samples
               Processing UNK/RNG Voucher Samples
           1)
    Following fixation for 5 to 7 days, decant and properly discard the formalin solution. Formalin is a
    potential carcinogen and should be used with extreme caution, as vapors and solution are highly
    caustic and may cause severe irritation on contact with skin, eyes, or mucus membranes. Wear vinyl or
    nitrile gloves and safety glasses, and always work in a well-ventilated area.
    a)   Formalin must be disposed of properly. Contact your regional EPA coordinator if your laboratory
        does not have the capability of handling waste formalin.
2)  Replace the formalin with tap water and soak specimens over a 4-5 day period. Soaking may require
    periodic water changes and should continue until the odor of formalin is barely detectable.
3)  Decant the tap water.  Use 45%-50% isopropyl alcohol or 70% ethanol as a final preservative for
    specimens.
4)  You will receive a  Fish  Update Identification and Count Form for each UNK/RNG sample from the NARS
    IM staff. This form lists all records from the original  collection form that were marked as being part of
    the UNK/RNG voucher sample. Identify unknown fish to species in the laboratory, using the procedure
    described in the NRSA laboratory operations manual, which is briefly described below.
    a)   Process unknowns by tag number—do not combine multiple bags (or jars) of the same unknown
        before determining the final identifications. Corrections and updates need to be linked back to
        the original page and line number, and voucher specimen tag you recorded on the collection
        form.
5)  Record the final identification(s) and count for each unknown  record in the Revised Name and  Revised
    Count columns of the update form.
    a)   Use common names from the standard list (Appendix D) as revised names.
    b)   If you must use a non-standard name, provide the page, line number, specimen tag number, and
        the taxonomic reference in the Comments section of the update form.
6)  If an unknown turns out to include > 1 species, correct the final counts based on the proportion of
    each species found in the original unknown bag.
    a)   Record the revised name and count for one species on the line of the original unknown.
    b)   Record the revised name and count for the second species as a new record on the update form.
        i)   Record the page, line number, and specimen tag number from the original unknown record
           on the next available blank line of the update form.
        ii)  Mark New Record Leave the Original Name and Original Count columns blank.
        iii) Record the revised name and count in the Revised Name and Revised Count columns.
7)  After reconciling all of the unknowns, submit the completed update forms to the NARS IM staff in
    Corvallis. Retain the preserved UNK/RNG voucher samples. Contact your regional  EPA coordinator if
    you cannot store the samples at your facility.
132

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           National Rivers and Streams Assessment 2013/14                             Field Operations Manual
           Version 1.0, May 2013                                                                 Wadeable

           Table 10.8 Procedure: Processing QA Voucher Samples

           1)   Ensure that all species collected at a site are represented by either preserved voucher specimens or
               photovouchers. There should be a unique QA voucher specimen tag assigned to every species
               recorded on the fish collection form.
           2)   Before submitting the QA Voucher sample, ensure that all specimens have been positively identified. If
               your attempts at identification do not yield a positive identification for 100% of the fish you retained,
               contact the Field Logistics Coordinator for further guidance (Chris Turner, cturner@glec.com, 715-829-
               3737).
           3)   After preparing the preserved QA voucher sample, check that the sample ID number recorded on the
               fish gear form matches the preprinted label attached to each sample jar, and that the number of jars
               recorded on the fish gear form is correct.
           4)   Retain the QA voucher samples in appropriate storage space for formalin until you receive information
               regarding where to send them from the NRSA staff at EPA Office of Water.
           5)   If you are storing the preserved QA voucher samples for an extended period, you  may need to replace
               the formalin fixative with ethanol.
               a)   Following fixation for 5 to 7 days, decant and properly discard the formalin solution. Formalin is a
                   potential carcinogen handle with extreme caution, as vapors and solution are highly caustic and
                   may cause severe irritation on contact with skin, eyes, or mucus membranes. Wear vinyl or nitrile
                   gloves and safety glasses, and always work in a well-ventilated area.
               b)   Formalin must be disposed of properly. Contact your regional EPA coordinator if your laboratory
                   does not have the capability of handling waste formalin.
           6)   Replace the formalin with tap water and soak specimens over a 4-5 day period. Soaking may require
               periodic water changes and should continue until the odor of formalin is barely detectable.
           7)   Decant the tap water. Use 45%-50% isopropyl alcohol or 70% ethanol as a final preservative for
               specimens.
           8)   When ready to ship all of the QA voucher samples, complete a sample tracking form as described in
               Appendix C.
           9)   Package the preserved samples properly for either formalin or ethanol and prepare all required
               documentation and safety measures for the shipment.
           10) Include a CD with the photovoucher files for each QA voucher sample in the shipment. Use the file
               names that are recorded on the fish gear form.

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National Rivers and Streams Assessment 2013/14                           Field Operations Manual
Draft Version 1.0, May 2013                                                         Wadeable
11     FISH TISSUE PLUG SAMPLING METHODS

11.1   Method Summary

Because many fish spend their entire life in a particular water body they can be important
indicators of water quality, especially for toxic pollutants (e.g., pesticides and trace elements).
Toxic pollutants, which may be present in the water column or sediments at concentrations
below our analytical detection limits, can  be found in fish tissue due to bioaccumulation.

Typical fish tissue collection methods require the fish to be sacrificed, whether it be a whole fish
or a skin-on fillet tissue sample. This can be problematic when there is a need to collect large
trophy-sized fish for contaminant analysis or when a large sample size is necessary for statistical
analysis. The following describes an alternative method for the collection of fish tissue samples
which uses a tissue plug instead of a skin-on fillet. Two fish tissue plugs for mercury analysis will
be collected from two fish of the same species (one plug per fish) from the target list (below) at
every site. These fish are collected during the fish assemblage sample collection effort (Section
10). A plug tissue sample is collected by inserting a biopsy punch into a de-scaled thicker area of
dorsal muscle section of a live fish. After collection, antibiotic salve is placed over the wound
and the fish is released.

11.2   Equipment and Supplies

Table 11.1 lists the equipment and supplies necessary for field crews to collect fish tissue plug
samples. This list is comparable to the checklist presented in Appendix A, which provides
information to ensure that field crews bring all of the required equipment to the site. Record
the fish tissue plug sampling data on the Fish Gear and Voucher/Tissue Sample Information
Form (Figure 10.6).
Table 11.1 Equipment and Supplies: Fish Tissue Plug Sample

 For fish tissue plug samples      Fish measuring board
                              Fish weigh scale
                              Plastic bags
                              Sterile 20  mL glass scintillation vial
                              Coolers with ice
                              Cooler with dry ice
                              Nitrile gloves                                                          y,
                              8 millimeter disposable biopsy punch (Acuderm brand Acu-Punch or            Q
                              equivalent)                                                           ^
                              Sterile disposable scalpel                                               ^
                              Sterile forceps                                                        ^
                              Laboratory pipette bulb.                                                z
                              Antibiotic salve.                                                       Q-
                              Fish collection gear (electrofisher, nets, livewell, etc.)                        ^
                              Dip net                                                               £
                              Field Operations Manual and laminated Quick Reference Guide                ID
 For recording measurements     Fish tissue plug sample labels                                            °-
                              Fish Gear and Sampling Form                                            ID
                                                                                                  to
                              Soft (#2) lead pencils for recording data on field forms                       ^
                              Fine-tipped indelible markers for filling out sample labels                     -,-
                              Clear tape strips for covering labels                                       •£


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          11.3   Sample Collection Procedures
          The fish tissue plug indicator samples will be collected using the same gear and procedures used
          to collect the fish assemblage. Collection of individual specimens for fish tissue occurs in the
          sample reach during the fish assemblage sampling. Samples should be taken from the species
          listed in the target list found in Table 11.2.  If the target species are unavailable, the fisheries
          biologist will select an alternative species (i.e., a species that is commonly consumed in the
          study area, with specimens of harvestable or consumable size) to obtain a sample from the
          species that are available. Recommended and  alternate target species are given in Table 11.2.
          The procedures for collecting and processing fish plug samples are presented in Table 11.3.
          Table 11.2 Recommended Target and Alternate Species for  Fish Tissue Plug Collection
             B







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Family name


Centrarchidae



Ictaluridae


Percidae

Moronidae
Esocidae


Salmonidae


Cyprinidae
Common name
Spotted bass
Largemouth bass
Smallmouth bass
Black crappie
White crappie
Channel catfish
Blue catfish
Flathead catfish
Sauger
Walleye
Yellow perch
White bass
Northern pike
Chain pickerel
Brown trout
Cutthroat trout
Rainbow trout
Brook trout
Northern pikeminnow
Scientific name
Micropterus punctulatus
Micropterus salmoides
Micropterus dolomieu
Pomoxis nigromaculatus
Pomoxis annularis
Ictalurus punctatus
Ictalurus furcatus
Pylodictis olivaris
Sander canadensis
Sander vitreus
Perca flavescens
Morone chrysops
Esox lucius
Esox niger
Salmo trutta
Oncorhynchus clarkii
Oncorhynchus mykiss
Salvelin us fan tin alis
Ptychocheilus oregonensis
Length Guideline
(Estimated
Minimum)
~280 mm
~280 mm
~300 mm
~330 mm
~330 mm
~300 mm
~300 mm
~300 mm
~380 mm
~380 mm
~330 mm
~330 mm
~430 mm
~430 mm
~300 mm
~300 mm
~300 mm
~330 mm
~300 mm
Centrarchidae
Bluegill

Rock bass

Redbreast sunfish
Lepomis macrochirus

Ambloplites rupesths

Lepomis auritus
"200 mm

"200 mm

"200 mm
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Table 11.3 Procedure: Fish Tissue Plug Samples
                                                                         Field Operations Manual
                                                                                      Wadeable
Fish Tissue Plug Methods
 1.
     Put on clean nitrile gloves before handling the fish. Do not handle any food, drink, sunscreen, or
     insect repellant until after the composite sample has been collected, measured, and wrapped.
 2.   Rinse  potential target species/individuals in ambient water to remove any foreign material from
     the external surface and  place in clean holding containers (e.g., livewells, buckets). Return non-
     target fishes or small specimens to the river or stream.
 3.   Retain two  individuals of  the same target species from each site. The fish should be of adequate
     size to sample (refer to Table 11.2 for minimum species length guidelines). Select fish based on the
     following criteria:
        •    is on the target list,
        •    both the same species
        •    both satisfy legal requirements of harvestable size (or weight) for the sampled river, or at
            least be of consumable size if no legal harvest requirements are in effect, and
        •    are of similar size,  so that the smaller individual is no less than 75% of the total length of
            the larger individual.
 4.   Remove one fish retained  for analysis from the clean holding container(s) (e.g., livewell) using clean
     nitrile gloves.
 5.   Measure the fish to determine total body length. Measure total length of the specimen in
     millimeters, from the anterior-most part of the fish to the tip of the longest caudal fin ray (when
     the lobes of the caudal fin are depressed dorsoventrally).
 6.   Weigh the fish in grams  using the fish weigh scale.
 7.   Note any anomalies (e.g.,  lesions, cuts, sores, tumors, fin erosion) observed on the fish.
 8.   Record site ID, date, sample ID, species, and specimen length and weight on the back of the Fish
     Gear and Sampling Form in the Fish Tissue Plug section (Figure 10.6). Make sure the sample ID
     numbers and specimen numbers/lengths that are recorded on the collection form match those on
     the sample tracking form and labels where applicable.
 9.   Prepare a Sample Identification Label for the sample, ensuring that the label information matches
     the information recorded  on the Fish Tissue Plug section of the Fish Gear and Sampling Form. Affix
     label to a sterile 20 milliliter scintillation vial and cover with clear tape.
 10.  On a  meaty portion of the left side dorsal area of the fish between the dorsal fin and the lateral
     line, clear a  small area of scales with a sterile disposable scalpel
 11.  Wearing clean nitrile gloves, insert the 8 millimeter biopsy punch into the dorsal muscle of the fish
     through the scale-free area. The punch  is inserted with a slight twisting motion cutting the skin and
     muscle tissue. Once full  depth of the punch is achieved a slight bending or tilting of the punch is
     needed to break off the end of the sample. Remove biopsy punch taking care to ensure sample
     remains in the punch. Note: The full depth of the punch should be filled with muscle tissue, which
     should result in collecting a minimum of 0.25 to 0.35 grams of fish tissue for mercury analysis.
 12.  Apply a generous amount  of antibiotic salve to the plug area and gently return the fish to the
     water.
 13.  Using a laboratory pipette bulb placed on the end of the biopsy punch, give a quick squeeze,
     blowing the tissue sample into a sterile  20 milliliter scintillation vial.
 14.  Repeat steps 2-13 for the second fish, collecting a second fish plug sample. Place the second plug in
     the same scintillation vial as the first. The two plugs should provide at least 0.5 grams of tissue.
 15.  Place the sample immediately on dry ice for shipment.
 16.  Dispose of gloves, scalpel and biopsy punch.
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            Fish Tissue Plug Methods
            17. Keep the samples frozen on dry ice or in a freezer at <-20°C until shipment
            18. Frozen samples will subsequently be packed on dry ice and shipped to the batched sample
                laboratory via priority overnight delivery service within 1 week.
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National Rivers and Streams Assessment 2013/14                           Field Operations Manual
Draft Version 1.0, May 2013                                                       Wadeable

12    WHOLE FISH SAMPLING METHOD

12.1  Method Summary
Fish are important integrators of toxic contaminants that are bioavailable in the water column
and in sediment. EPA monitors the occurrence of toxic chemicals in fish fillet samples to assess
the potential health impacts to people who consume fish. Results from the 2008/09 NRSA
provided the first statistically representative national data for fish contamination in U.S. rivers.
Collecting whole fish tissue samples and submitting them to the laboratory for filleting and
homogenization during the 2013/14 NRSA allows consistency with fish tissue methods of the
previous NRSA, and provides tissue amounts that are sufficient to allow the analysis of multiple
chemical contaminants of concern. It will also provide the first opportunity for temporal analysis
of probability-based national fish contamination trends in U.S. rivers. Additionally, collecting fish
at locations sampled during the previous NRSA will reduce the variability in data for trends
analysis.
Whole fish tissue sampling procedures are described in detail in Table 2.3. The objective is to
collect one whole fish tissue sample from each of the 453 designated target sites. The focus is
on fish species that commonly occur throughout the region of interest and that are sufficiently
abundant within a sampling reach. Each whole fish tissue sample will consist of five adult fish of
the same species that are similar in size (the smallest individual in the sample is no less than
75% of the total length of the largest individual). Collection occurs in the sampling reach. Whole
fish samples are shipped to the laboratory designated for fish sample preparation. The
laboratory fillets the fish and homogenizes the fillet tissue for analysis of mercury and other
contaminants (e.g., perfluorinated compounds).

12.2  Equipment and Supplies
Table 11.4 lists the equipment and supplies necessary for field crews to collect whole fish tissue
samples. This list is comparable to the checklist presented in  Appendix A, which provides
information to ensure that field crews bring all of the required equipment to the site. Record the
fish tissue sampling data on the Whole Fish Tissue Collection Form (Figure 11.1).
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          Table 11.4 Equipment and Supplies: Whole Fish Tissue Sample Collection
            For collecting whole
            fish tissue sample
            For storing and
            preserving whole
            fish tissue sample
            For documenting the
            whole fish tissue
            sample

            For shipping the
            whole fish tissue
            samples
                              Electrofishing equipment (including
                              variable voltage pulsator unit, wiring
                              cables, generator, electrodes, dip nets,
                              protective gloves, boots, and necessary
                              safety equipment)
                              Scientific collection permit
                              Sampling vessel (including boat, motor,
                              trailer, oars, gas, and all required safety
                              equipment)
                              Aluminum foil (solvent rinsed and
                              baked)
                              Heavy-duty food grade polyethylene
                              tubing
                              Large plastic (composite) bags
                              Whole Fish Tissue Collection Form
                              Clipboard
                              Clear tape strips

                              Preaddressed FedEx airbill
                              Coolers
Coast Guard approved personal
floatation devices
Maps of target sites & access
routes Global Positioning System
(GPS) unit
Livewell and/or buckets
Measuring board (millimeter
scale) Clean nitrile gloves
Knife or scissors
Dry Ice
Plastic cable ties
Coolers

Sample Identification Labels
#2 pencils
Fine tipped indelible markers
Tracking Form
Chain-of-custody labels
Packing/strapping tape
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           12.3   Sampling Procedures
           The whole fish tissue samples will be collected using the same gear and procedures used to
           collect the fish assemblage samples. Collection of individual specimens for whole fish samples
           occurs in the sample reach during the fish assemblage sampling. Ideally, each fish sample will
           contain 5 fish of the same species that are similar in size. Depending on the size of the fish,
           fewer than 5 fish may be acceptable or more than 5 fish will be necessary to meet the 500-gram
           tissue requirement for chemical analysis and archived tissue (refer to Frequently Asked
           Questions  in the sampling kits). If sufficient fish are not  collected during the fish assemblage
           sampling, sample for up to one additional hour. If no fish can be collected, call the Field  Logistics
           Coordinator at the end  of the day and  record "no sample collected" on the whole fish tissue
           collection form, along with the reason in the comments section of the form. If the target species
           are unavailable, the fisheries biologist will select an alternative species (i.e., a species that is
           commonly consumed in the study area, with specimens of harvestable or consumable size, and
           in sufficient numbers to yield a fish sample with adequate tissue for analysis) to obtain a whole
           fish sample from the species that are available. Recommended target species are given in Table
           11.5. The procedures for collecting and processing whole fish tissue samples are presented in
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Table 11.6.
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Table 11.5 Recommended Target Species for Whole Fish Tissue Collection
                                                                                 Field Operations Manual
                                                                                              Wadeable
^^^H Family name Common name Scientific name
Target Species
Centrarchidae
Ictaluridae
Percidae
Moronidae
Esocidae
Salmonidae
Spotted bass
Largemouth bass
Smallmouth bass
Black crappie
White crappie
Channel catfish
Blue catfish
Flathead catfish
Sauger
Walleye
Yellow perch
White bass
Northern pike
Chain pickerel
Brown trout
Cutthroat trout
Rainbow trout
Brook trout
Micropterus punctulatus
Micropterus salmoides
Micropterus dolomieu
Pomoxis nigromaculatus
Pomoxis annularis
Ictalurus punctatus
Ictalurus furcatus
Pylodictis olivaris
Sander canadensis
Sander vitreus
Perca flavescens
Moron e chrysops
Esox lucius
Esox niger
Salmo trutta
Oncorhynchus clarkii
Oncorhynchus mykiss
Salvelin us font in alis
~280 mm
~280 mm
~300 mm
~330 mm
~330 mm
~300 mm
~300 mm
~300 mm
~380 mm
~380 mm
~330 mm
~330 mm
~430 mm
~430 mm
~300 mm
~300 mm
~300 mm
~330 mm
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Table 11.6 Sampling Procedures for Whole Fish Tissue Samples
 Whole Fish Tissue Method
 1.
    Put on clean nitrile gloves before handling the fish. Do not handle any food, drink, sunscreen, or
    insect repellant until after the whole fish sample has been collected, measured, and wrapped.
2.   Rinse potential target species/individuals in ambient water to remove any foreign material from the
    external surface and place in clean holding containers (e.g., livewells, buckets). Return non-target
    fishes or small specimens to the river or stream.
3.   Collect one target species sample from each designated site. The sample should consist of 5 fish of
    adequate size to provide a total of 500 grams of edible tissue for analysis (refer to Table 11.5 for
    minimum species length guidelines). Select fish  for each sample based on the following criteria:
        •   all are of the same species,
        •   all satisfy legal requirements of harvestable size (or weight) for the sampled river, or at
            least be of consumable size if no legal harvest requirements are in effect,
        •   all are of similar size, so that the smallest individual in a composite is no less than 75% of
            the total length of the largest individual, and
        •   all are collected at the same time, i.e.,  collected as close to the same time  as possible, but
            no more than one week apart (Note: Individual fish may have to be frozen until all fish to
            be included in the sample are available for delivery to the designated  laboratory).
        Accurate taxonomic identification is essential in assuring and defining the  organisms that have
        been collected and submitted for analysis.  Do not mix species in a single whole fish sample.
4.   Measure each individual fish to determine total body length.  Measure total length of each
    specimen in millimeters, from the anterior most part of the fish to the tip of the longest caudal fin
    ray (when the lobes of the caudal fin are depressed  dorsoventrally).
5.   Record site ID, date, sample ID, species (common name), and specimen length  on the Whole Fish
    Tissue Collection Form (Figure 11.1) in black ink. Fill in site type ("Wadeable" or "Boatable") at the
    top of the form.  Address the two sample criteria in the space above the fish specimen data to
    confirm compliance. All samples must meet these two criteria (i.e., fish are all the same species and
    fish lengths are all within 75% of the largest specimen length). Make sure the sample ID numbers
    and specimen numbers/length s that are recorded on the collection form match the corresponding
    information on each individual specimen label.
6.   Remove each fish selected for analysis from the clean holding container(s) (e.g., livewell)  using
    clean nitrile gloves. Dispatch each fish using a clean  wooden bat (or equivalent wooden device).
7.   Wrap each fish in extra heavy-duty aluminum foil with the dull  side in (foil provided by EPA as
    solvent-rinsed, oven-baked sheets).
8.   Prepare  a Sample Identification Label for each sample, ensuring that the label information matches
    the information  recorded on the Whole Fish Tissue Collection Form.
9.   Cut a length of food grade tubing (provided by EPA) that is long enough to contain each individual
    fish and to allow extra length on each end to secure with cable ties. Place each foil wrapped
    specimen into the appropriate length of tubing. Seal each end of the tubing with  a plastic cable tie.
    Attach the fish sample label to the outside of the food grade tubing with clear tape  and secure the
    label by taping around the entire fish  (so that tape sticks to tape).
10. Place all the wrapped fish in the sample from each site in a large plastic bag and seal with another
    cable tie.
11. After each sample is packaged, place it immediately on dry ice for shipment.  If samples will be
    carried back to a laboratory or other facility to be frozen before shipment, wet ice can be used to
    transport wrapped and bagged fish samples in the coolers to a  laboratory or other interim facility.
12. If possible, keep all (five) specimens designated for a particular sample in the same  shipping
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            Whole Fish Tissue Method
               container (ice chest) for transport.
            13. Samples may be stored temporarily on dry ice (replenishing the dry ice daily). You have the option,
               depending on site logistics, of:
                    •   shipping the samples packed on dry ice in sufficient quantities to keep samples frozen for
                       up to 48 hours (50 pounds are recommended), via priority overnight delivery service (e.g.,
                       Federal Express), so that they arrive at the sample preparation laboratory within less than
                       24 hours from the time of sample collection, or
                    •   freezing the samples within 24 hours of collection at <-20°C, and storing the frozen
                       samples until shipment within 2 weeks of sample collection (frozen samples will
                       subsequently be packed on dry ice and shipped to the sample preparation laboratory via
                       priority overnight delivery service).
            14. Ship fish tissue samples to the designated laboratory for fish sample preparation on  Monday
               through Thursday (no Saturday delivery to the laboratory).
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                               NRSA 2013/14 WHOLE FISH TISSUE COLLECTION
                                          O WADEABLE     O BOATABLE
                                                                                      Reviewed by i.ir.tui],_
    WHOLE FISH TISSUE FILLET SAMPLE (FTIS)   SAMPLE ID:
                                                                                      NO SAMPLE COLLECTED Q
           O FISH ARE ALL THE SAME SPECIES
O FISH ALL WITHIN 75% OF LARGESTSPECIMEN
     .01
     .02
     .03
     .04
     .05
     .06*
     .07*

     .08*
     .09*

     .10*
          Common Name
                                                    Total
                                                 Length (mm)
     O
     O
     O
     O
     O
     O
     'Additional specimens for smaBer fish species to ensure sufficient [issue is available for chemical analysis of filet tissue
         04/08/2013  2013 Fish Tissue Collection
                                                                                         2227506532
Figure 11.1 Whole Fish Tissue Collection Form
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          13    FINAL SITE ACTIVITIES

          13.1  Overview of Final Site Activities
          Prior to leaving the site, make a general visual assessment of the site and its surrounding
          catchment. The objective of the site assessment is to record observations of catchment and site
          characteristics that are useful for future data interpretation, ecological value assessment,
          development of associations, and verification of stressor data. Your observations and
          impressions are extremely valuable.
          You will filter and process the fecal indicator, chlorophyll-a, and periphyton samples, as well as.
          Conduct a final check of the data forms, labels and samples. The purpose of the second check of
          data forms, labels and samples is to assure completeness of all sampling activities. Finally, clean
          and pack all equipment and supplies, and clean the launch site and staging areas. After you
          leave the site, report the sampling event to the Information Management Coordinator, and ship
          or store the samples.  Activities described in this section are summarized in Figure 13.1.
                                           COMPLETE SITE
                                            ASSESSMENT
                                              (4 People)
                      REVIEW DATA FORMS
                          (Crew Leader)
                     •  Completeness
                     •  Accuracy
                     •  Legibility
                     •  Flags/Comments
  FILTER, PRESERVE, &
    INSPECT SAMPLES
       (3 People)
 •   Complete
 •   Sealed
 •   Ice packs
 •   Packed for transport
                     REVIEW SAMPLE LABELS
                          (Crew Leader)
                    • Completeness
                    • Accuracy
                    • Legibility
                    • Cross-check with forms
  INSPECT BOAT, MOTOR,
  TRAILER, AND NETS FOR
 PRESENCE OF PLANT AND
  ANIMAL MATERIAL, AND
   CLEAN THOROUGHLY
        (3 People)
                      PACK EQUIPMENT AND
                    SUPPLIES FOR TRANSPORT
                           (2 People)
LOAD BOAT ONTO TRAILER;
 CLEAN UP LAUNCH SITE
   AND STAGING AREA
        (2 People)
                                            LEAVE SITE
                                         COMMUNICATIONS
          SHIP SAMPLES
          Figure 13.1 Final Site Activities Summary
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13.2   General Site Assessment
Complete the Site Assessment Form (Figure 13.2) after sampling, recording all observations
from the site that were noted during the course of the visit. This Site Assessment Form is
designed as a template for recording pertinent field observations. It is by no means
comprehensive, and any additional observations should be recorded in the General Assessment
section.

13.2.1  Elevation at Transect K
Record elevation at Transect K using your GPS device. To record this information, record the
elevation holding the GPS at approximately 3 feet above the surface of the water. Ensure that
the numbers are properly recorded for Transect K on the Assessment Form.

13.2.2  Watershed Activities and Disturbances Observed
Record any of the sources of potential stressors listed in the "Watershed Activities and
Disturbances Observed" section on the Site Assessment Form (Figure 13.2). Include those that
were observed while on the site, while driving or walking through the site catchment, or while
flying over the site and catchment. For activities and stressors that you observe, rate their
abundance or influence as low (L), moderate (M), or heavy (H) on the line next to the listed
disturbance. Leave the line blank for any disturbance not observed. The distinction between
low, moderate, and heavy will be  subjective. For example, if there are two to three houses on a
site, circle "L" for low next to "Houses." If the site is ringed with houses, rate it as heavy (H).
Similarly, a small patch of clear-cut logging on a hill overlooking the site would rate a low
ranking. Logging activity right on the site shore, however, would get a heavy disturbance
ranking. This section includes residential, recreational, agricultural, industrial, and stream
management categories.

13.2.3  Site Characteristics
Record observations regarding the general characteristics of the site on the Site Assessment
Form (Figure 13.2 & Figure 13.3).  When assessing these characteristics, look at a 200 m riparian
distance on both banks. Rank the  site between "pristine" and  "highly disturbed", and between
"appealing" and "unappealing." Document any signs of beaver activity and flow modifications.
Record the dominant land use and forest age class. Document the weather conditions on the
day of sampling, and any extreme weather conditions just in the days prior to sampling.

13.2.4  General Assessment
Record any additional information and observations in this narrative section. Information to
include could be observations on biotic integrity, vegetation diversity, presence of wildlife, local
anecdotal information, or any other pertinent information about the site or its catchment.
Record any observations that may be useful for future data interpretation.
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LJJ
H
>
                                                      NRSA 2013/U ASSESSMENT (Front)
                                                                              Date;         /
                                                                                                        R«vl*w*dby(.r*ltJil):_
                   Elevation at transect K:
                                                             Oft  Om
                   WATERSHED ACTIVITIES AND DISTURBANCES OBSERVED
                   (Intensity: Blank^Not observed, l_ = l_ow. M=Moderate, H=HeavyJ
                                                                                           BLANK FIELD INDICATES ABSENCE
                                                        o
                   Residential
                   O O O R«si*»»«5
                   O 0 O Manila inedLawis
                   © 0 © Construction
                   O O O P'P's. Drains
                   O O O Dumping
                   O 0 O RMl"
                   O 0 O Bridges/Causeway
                   O 0 O Sewsgs Treatment
                                        Recreational
                                      O O O HikingTiads
                                      O © O Parks.Cnmpgrounds
                                      O 0 0 Prinii'r*
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     Site ID:
                                      NRSA 2013/14 ASSESSMENT (Back)

                                                            Date:        !         I
    GENERAL ASSESSMENT AND COMMENTS
    INVASIVE OR NUISANCE SPECIES OF LOCAL INTEREST
    Record species of plants and animals that were observed but are not on the invasive plant form. Examples would be Zebra Mussel or
    New Zealand Mud Snail, or invasive plants or animals of concern to a particular state. Indicate your level of confidence in your
    identification, and provide some idea of how prevalent it is in the sampling reach or adjacent riparian area.
      Species (Common Name)
                                   Confidence
                                                  Prevalence
                                                                      Comments
                                     OLOW
                                     Q HIGH
O DOMINANT O SPARSE
Q COMMON	
                                     OLOW
                                     O HIGH
O DOMINANT O SPARSE
O COMMON
                                     OLOW
                                     O HIGH
O DOMINANT
O COMMON
OSPARSE
                                     OLOW
                                     O HIGH
O DOMINANT O SPARSE
O COMMON
                                     O LOW
                                     O HIGH
O DOMINANT
O COMMON
                                     O LOW
                                     OHIGH
O DOMINANT O SPARSE
O COMMON
                                     OLOW
                                     O HIGH
O DOMINANT
O COMMON
O SPARSE
                                     OLOW
                                     OHIGH
O DOMINANT O SPARSE
O COMMON
         04/08/2013  2013 Assessment
                                                                                         0986596801
Figure 13.3 Site Assessment Form (back)
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           13.3   Processing the Fecal Indicator, Chlorophyll-a, and Periphyton Samples
           13.3.1 Equipment and Supplies (Fecal Indicator Filtering)
           Table 13.1 provides the equipment and supplies needed for field crews to collect the fecal
           indicator sample.
           Table 13.1 Equipment and Supplies: Fecal Indicator Sample Processing
             For processing samples
             For recording measurements
                                Nitrile gloves
                                sterile screw-cap 50-mL PP tube
                                Filtration apparatus with collection flask
                                Sterile filter holder, Nalgene 145/147
                                Vacuum pump (electric pump may be used if available)
                                Sterile phosphate buffered saline (PBS)
                                Osmotics 47 mm polycarbonate sterile filters
                                Sterile disposable forceps
                                Petri dishes (60 x 15, disposable)
                                2 sterile microcentrifuge tubes containing sterile glass beads
                                1 additional sterile microcentrifuge tube if collecting filter blank
                                Dry ice
                                Cooler
                                Field Operations Manual and laminated Quick Reference Guide
                                Sample Collection Form
                                Soft (#2) lead pencils for recording data on field forms
                                Fine-tipped indelible markers for filling out sample labels
                                Fecal Indicator sample labels (2 vial labels and 1 bag label)
                                Filter blank label if collecting filter blank
                                Clear tape strips for covering labels
           13.3.2 Procedures for Processing the Fecal Indicator Sample
           The fecal indicator sample must be filtered before the chlorophyll-o and periphyton samples,
           since the filtering apparatus needs to be sterile for this sample. The procedures for processing
           the fecal  indicator sample are presented in Table 13.2. The sample must be filtered and frozen
           within 6 hours of collection.
           Table 13.2 Procedure: Fecal Indicator Sample Processing
1.   Put on nitrile gloves.
2.   Set up sample filtration apparatus on flat surface and attach vacuum pump. Set-out 50-mL sterile PP
    tube, sterile 60-mm Petri dish, 2 bottles of chilled phosphate buffered saline (PBS), Osmotics 47 mm
    polycarbonate sterile filter box, and 2 filter forceps.
3.   Chill Filter Extraction tubes with beads on dry ice.
4.   Aseptically transfer 2 polycarbonate filters from filter box to base of opened Petri dish. Close filter
    box and set aside.
5.   Remove the pre-loaded cellulose nitrate (CN) filter (the filter with grid design on it) from funnel and
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    discard. Be sure to leave the support pad in the filter funnel.
6.   Load filtration funnel with sterile polycarbonate filter on support pad (shiny side up).
7.   Shake sample bottle(s) 25 times to mix well.
8.   Measure 25-mL of the mixed water sample in the sterile graduated PP tube and pour into filter
    funnel.
9.   Replace cover on filter funnel and pump to generate a vacuum (do not generate more than 7 inches
    of Hg of pressure). Keep pumping until all liquid is in filtrate collection flask.
10. If the first 25 mL volume passes readily through the filter, add another 25 mLand continue filtration.
    If the filter clogs before completely filtering the first or second 25 mL volume, discard the filter and
    repeat the filtration  using a lesser volume.
11. Pourapprox. 10-mLof the chilled phosphate buffered saline (PBS) into the graduated PP tube used
    for the sample. Cap the tube and shake 5 times. Remove the cap and pour rinsate into filter funnel to
    rinse filter.
12. Filter the rinsate and repeat with another 10 mL of phosphate buffered saline (PBS).
13. Remove filter funnel from base without disturbing filter. Using sterile disposable forceps remove the
    filter (touching only the filter edges) and fold it in half, in quarters, in eighths, and then in sixteenths
    (filter will be folded 4 times).
14. Insert filter into chilled filter extraction tube (with beads). Filter should be inserted open end down
    (pointed side up) into the tube. Replace and tighten the screw cap, insert tube(s) into ziplock bag on
    dry ice for preservation during transport and shipping.
15. Record the volume of water sample filtered through each filter and the volume of buffer rinsate each
    filter was rinsed with on the Sample Collection Form, Side 2. Record the filtration start time and finish
    time for each sample.
        Repeat steps 6 to 15 for the remaining 50-mL sub-sample volume to be filtered.
Processing Procedure—fecal indicator filter blank
    Enterococci filter blanks will be prepared at all revisit sites during the first visit. Prepare the filter
    blanks before filtering the river sample.
    1.   Set up sample filtration apparatus using same procedure as used for the river sample. Chill Filter
        Extraction tubes with beads on dry ice.
    2.   Aseptically transfer 1 polycarbonate filter from filter box to base of opened Petri dish. Close filter
        box and set aside.
    3.   Remove the pre-loaded cellulose nitrate (CN) filter (the filter with grid design on it) from funnel
        and discard. Be sure to leave the support pad in the filter funnel.
    4.   Load filtration funnel with sterile polycarbonate filter on support pad (shiny side up).
    5.   Measure 10-mLof the chilled phosphate buffered saline  (PBS) in the sterile graduated PP tube
        and pour into filter funnel.
    6.   Replace cover on filter funnel and pump to generate a vacuum (do not generate more than 7
        inches of Hg of pressure). Keep pumping until all liquid is in filtrate collection flask.
    7.   Remove filter funnel from base without disturbing filter.  Using sterile disposable forceps remove
        the filter (touching only the filter edges) and fold it in half,  in quarters, in eighths, and then in
        sixteenths (filter will be folded 4 times).
    8.   Insert filter into chilled filter extraction tube (with beads). Filter should be inserted open end
        down (pointed side up) into the tube. Replace and tighten the screw cap, insert tube(s) into
        ziplock bag on dry ice for preservation during transport and shipping.
    9.   Label the samples as "blank" on the label and field form, and package and submit this sample to
        the lab with the standard samples.
    10. Indicate that you have collected a filter blank by filling in the "Blank Collected" button on the
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                  Sample Collection Form.
           13.3.3 Equipment and Supplies (Chlorophyll-a from Water Sample Filtering)
           Table 13.3 provides the equipment and supplies needed to process the chlorophyll-a water
           sample.
           Table 13.3 Equipment and Supplies:r Chlorophyll-a Processing
           For filtering chlorophyll-a sample
                                Whatman GF/F 0.7 u.m glass fiber filter
                                Filtration apparatus with graduated filter holder and collection flask
                                Vacuum pump (electric pump may be used if available)
                                50-mL screw-top centrifuge tube
                                Aluminum foil square
                                250 mLgraduated cylinder
                                Dl water
                                Nitrile gloves
                                Forceps
                                Dry ice
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           For recording measurements
                                Sample Collection Form
                                Sample labels
                                #2 pencils
                                Fine-tipped indelible markers
                                Clear tape strips
           13.3.4 Procedures for Processing the Chlorophyll-a Water Sample
           The procedures for processing chlorophyll-a water samples are presented in Table 13.4.
           Whenever possible, sample processing should be done in subdued light, out of direct sunlight.
           Table 13.4 Procedure: Chlorophyll-a Sample Processing
           Filtering for the chlorophyll a water sample
1.  Put on nitrile gloves.
2.  Use clean forceps to place a Whatman GF/F 0.7 urn glass fiber filter in the graduated filter holder
   apparatus with the gridded side of the filter facing down.
3.  Retrieve the 2 liter chlorophyll sample bottle from the cooler and shake the bottle to homogenize the
   sample. While filtering sample, keep the bottle in the cooler on ice.
4.  Measure 250 mLof water with a graduated cylinder and pour into the filter holder, replace the cap,
   and use the vacuum pump to draw the sample through the filter (do not exceed 7 inches of Hg).  If
   250 mL of site water will not pass through the filter, change the filter, rinse the apparatus with Dl
   water, and repeat the procedures using 100-mLof site water.
      •    NOTE: IF the water is green or turbid, use a smaller volume to start with.
5.  Observe the filter for visible color. If there is visible color, proceed; if not, repeat steps 3 & 4 until
   color is visible on the filter or until a maximum of 2,000 mL have been filtered. Record the actual
   sample volume filtered on the Sample Collection Form.
6.  Rinse the upper portion of the filtration apparatus and graduated cylinder thoroughly with Dl water
   to include any remaining cells adhering to the sides and pump through the filter. Monitor the level of
   water in  the lower chamber to ensure that it does not contact the filter or flow into the pump.
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For preparing
periphyton samples
    Remove the bottom portion of the apparatus and pour off the water from the bottom as often as
    needed.
 7.  Remove filter funnel from base without disturbing filter.
 8.  Remove the filter from the holder with clean forceps. Avoid touching the colored portion of the filter.
    Fold the filter in half, with the colored side folded in on itself.
 9.  Place the folded filter into a 50-mL screw-top centrifuge tube and cap. Tighten the cap as tightly as
    possible. The cap will seal tightly after an additional % turn past the point at which initial resistance is
    met. Failure to tighten the lid completely could allow water to infiltrate into the sample and may
    compromise its integrity.
 10. Record the sample volume filtered on a chlorophyll label and attach it to the centrifuge tube (do not
    cover the volume markings on the tube). Ensure that all written information  is complete and legible.
    Cover with a strip of clear tape.
11.   Wrap the tube in aluminum foil and place in a self-sealing plastic bag. Place this bag immediately on
     dry ice to freeze.

13.3.5 Equipment and Supplies (Periphyton Sample)
Table 13.5 lists the equipment and supplies needed to process the periphyton sample.
Table 13.5 Equipment and Supplies: Periphyton Samples
                       Whatman 47 mm 0.7 micron GF/F glass fiber filter
                       Whatman 47 mm 1.2 micron GF/C glass fiber filter
                       Filtration apparatus with collection flask  and graduated filter holder
                       Vacuum pump (electric pump may be used)
                       25 or 50-mL graduated cylinder
                       Pipette and pipette bulb (2 mL)
                       3 50 mL screw-top centrifuge tubes
                       60-mL syringe with tip removed
                       Aluminum foil squares
                       Forceps
                       deionized water in wash bottle
                       plastic electrical tape
                       dry ice
                       wet ice
                       coolers
                       formalin
                       Sample Collection Form
                       Sample labels
                       Pencils
                       Fine-tipped indelible markers
                       Clear tape strips

13.3.6 Procedures for Processing the Periphyton Samples
Three different types of laboratory samples are prepared from the composite index samples: an
ID/enumeration sample (to determine taxonomic composition and relative abundances),
chlorophyll a sample, and a biomass sample (for ash-free dry mass [AFDM]). All the sample
For data recording
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          containers required for an individual site should be sealed in plastic bags until use to avoid
          external sources of contamination (e.g., dust, dirt, or mud) that are present at site shorelines.

          13.3.6.1 ID/Enumeration Sample
          Prepare the ID/Enumeration sample as a 50-mL aliquot from the composite index sample,
          following the procedure presented in Table 13.6. Preserve each sample with formalin. Record
          the sample ID number from the container label and the total volume of the periphyton sample
          in the appropriate fields on the Sample Collection Form as shown in Error! Reference source not
          found.. Store the preserved samples upright in a  container containing absorbent material.
          Table 13.6 Procedure: ID/Enumeration Samples of Periphyton
          Periphyton ID Sample Processing Procedure
  1.   Prepare a sample label (with pre-printed sample ID number sample type "PERI"). Record the
      volume of the subsample (typically 50 mL) and the volume of the composite index sample on the
      label. Attach completed label to a 50-mL centrifuge tube; avoid covering the volume graduations
      and markings. Cover the label completely with a clear tape strip.
  2.   Record the sample ID number of the label and the total volume of the composite index sample on
      the Sample Collection Form.
  3.   Rinse a 60-mL syringe with deionized water.
  4.   Thoroughly mix the bottle containing the composite sample.
  5.   Immediately after mixing, withdraw 50 mLof the mixed sample into the syringe and place the
      contents of syringe into the labeled 50-mL centrifuge tube.
  6.  Use a syringe or bulb pipette to add 2ml of 10% formalin to the tube. Cap the tube tightly and  seal
      with plastic electrical tape. Tighten the cap as tightly as possible. The cap will seal tightly after an
      additional % turn past the point at which initial resistance is met.
  7.   Shake gently to distribute preservative.
  8.   Record the volume of the sample in the centrifuge tube (excluding  the volume of preservative) in
      "Assemblage ID Subsample Vol." field of the Sample Collection Form.

13.3.6.2Periphyton Chlorophyll a Sample
Prepare the periphyton chlorophyll a sample by filtering a 25-mL aliquot of the composite index
sample through a 47  mm 0.7 micron GF/F glass fiber filter. The procedure for preparing
periphyton chlorophyll a samples is presented in Table 13.7. Chlorophyll a can  degrade rapidly
when exposed to bright light. If possible, prepare the  samples in subdued light (or shade),
filtering as  quickly as possible after collection to minimize degradation. If using the same
filtration chamber that was used for Enterococci and index site chlorophyll-o samples, rinse it
with deionized water prior to filtering the periphyton  chlorophyll-o sample. If you are reusing a
filtration chamber from a previous site, you  should rinse it with Dl water each day before use at
the base site and then seal in a plastic bag until use at the stream (be sure to use a new chamber
at each site for the Enterococci sample as it  needs to be filtered in  a sterile chamber). Keep the
glass fiber filters in a dispenser inside a sealed plastic  bag until use.
It is important to measure the volume of the sample being filtered  accurately (±1 ml) with  a
graduated cylinder. During filtration, do not exceed 7  inches of Hg to avoid rupturing cells.  If the
vacuum pressure exceeds 7 inches of Hg, prepare a new sample. If  the filter clogs completely
before all the sample in the chamber has been filtered, discard the sample and filter, and
prepare a new sample using a smaller volume of sample.
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Table 13.7 Procedure: Preparing Chlorophyll Samples of Periphyton
Periphyton Chlorophyll a Sample Processing Procedure
    1.   Using clean forceps, place a Whatman GF/F 0.7 urn glass fiber filter on the filter holder gridded
        side down. If needed, use a small amount of deionized water from a wash bottle to help settle
        the filter properly. Attach the filter funnel to the filter holder and filter chamber, and then attach
        the vacuum pump to the filter flask.
    2.   Rinse the sides of the filter funnel and the filter with a small volume of deionized water to
        prevent cross-contamination from the  previously filtered sample.
    3.   Rinse a 25-mL or 50-mL graduated cylinder three times with small volumes of deionized water
        and discard.
    4.   Mix the composite sample bottle thoroughly.
    5.   Measure 25 mL (±1 mL) of sample into the graduated cylinder.
         • NOTE: For a composite sample containing fine sediment, allow grit to settle for 10 - 20
        seconds before pouring the sample into the  graduated cylinder.
    6.   Pour the 25-mL aliquot into the filter funnel, replace the cap, and pull the sample through the
        filter using the vacuum pump. Vacuum pressure from the pump should not exceed 7 inches of Hg
        to avoid rupture of fragile algal cells.
         • NOTE: If 25 mL of sample will not pass through the filter, discard the filter and rinse the
        chamber thoroughly with deionized water. Collect a new sample using a smaller volume of
        sample, measured to ±1 mL. Be sure to record the actual volume sampled on the sample label
        and the Sample Collection Form.
    1.   Monitor the level of water in the lower chamber to ensure that it does not contact the filter or
        flow into the pump.  Remove the bottom portion of the apparatus and pour off the water from
        the bottom as often as needed.
    8.   Remove the filter chamber from the filter holder being careful not to disturb  the filter. Remove
        the filter from the holder with clean forceps. Avoid touching the colored portion of the filter.
        Fold the filter in half, with the colored sample (filtrate) side folded in on itself. Place  the folded
        filter in a 50 mL centrifuge tube.
    9.   Tighten the cap as tightly as possible. The cap will seal tightly after an additional % turn past the
        point at which initial resistance is met.
    10.  Prepare a sample label (with pre-printed sample ID number sample type "PCHL") including the
        volume filtered, and attach it to the centrifuge tube. Cover the label completely with a strip of
        clear tape.
    11.  Place the centrifuge tube into a self-sealing plastic bag.
    12.  Record the sample ID number of the label and the total volume of the composite index sample
        on the Sample Collection Form. Record the volume filtered in the "Periphyton Chlorophyll" field
        on the Sample Collection Form. Double check that the volume recorded on the collection form
        matches the total volume recorded on the sample label.
    13.  Place the centrifuge tube containing the filter on dry ice.

13.3.6.3Periphyton Biomass Sample (AFDM)

Prepare the ash-free dry mass  (AFDM) sample by filtering a 25-mL aliquot of the composite
index sample through a 47 mm 1.2 micron GF/C glass fiber filter. The procedure for preparing
AFDM samples is presented in Table 13.8. If using the same filtration chamber that was used for
Enterococci and chlorophyll-a samples, rinse it with deionized water prior to filtering the
periphyton biomass sample. If you are reusing a filtration chamber from a previous site, you
should rinse it with Dl water each day before use at the base site and then seal in a plastic bag
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           until use at the stream (be sure to use a new chamber at each site for the Enterococci sample as
           it needs to be filtered in a sterile chamber). Keep the glass fiber filters in a dispenser inside a
           sealed plastic bag until use.

           It is important to measure the volume of the sample being filtered accurately (±1 mL) with a
           graduated cylinder. During filtration, do not exceed 7 inches of Hg to avoid rupturing cells. If the
           vacuum pressure exceeds 7 inches of Hg prepare a new sample. If the filter clogs completely
           before all the sample in the chamber has been filtered, discard the sample and filter, and
           prepare a  new sample using a smaller volume of sample.

           Table 13.8 Procedure: Preparing Ash-Free Dry Mass (AFDM) Samples of Periphyton
           Periphyton AFDM Sample Processing Procedures
1.
2.

3.

4.
5.
1.
               8.
                  Using clean forceps, place a Whatman 47 mm 1.2 micron GF/C glass fiber filters on the filter
                  holder gridded side down. If needed, use a small amount of deionized water from a wash bottle
                  to help settle the filter properly. Attach the filter funnel to the filter holder and filter chamber,
                  then attach the hand vacuum pump to the filter flask.
                  Rinse the sides of the filter funnel and the filter with a small volume of deionized water to
                  prevent contamination from the previously filtered sample.
                  Rinse a 25-mL or  50-mL graduated cylinder three times with small volumes of deionized water
                  and discard.
                  Mix the composite sample bottle thoroughly.
                  Measure 25 mL (±1 mL) of sample into the graduated cylinder.
                       •   NOTE: For a composite sample containing fine sediment, allow grit to settle for 10 - 20
                           seconds before pouring the sample into the graduated cylinder.
                  Pour the 25-mL aliquot into the filter funnel, replace the cap,  and pull the sample through the
                  filter using the vacuum pump. Vacuum pressure from the pump should not exceed 7 inches of Hg
                  to avoid rupture of fragile algal cells.
                       •   NOTE : If 25 mL of sample will not pass through the filter, discard the filter and rinse the
                           chamber thoroughly with deionized water. Collect a new sample using a smaller
                           volume of sample, measured to ±1 mL Be sure to record the actual volume sampled on
                           the sample label and the Sample Collection Form.
                  Monitor the level of water in the lower chamber to ensure that it does not contact the filter or
                  flow into the pump. Remove the bottom portion of the apparatus and pour off the water from
                  the bottom as often as needed.
    Remove the filter chamber from the filter holder being careful not to disturb the filter. Remove
    the filter from the holder with clean forceps. Avoid touching the colored portion of the filter.
    Fold the filter in half, with the colored sample (filtrate) side folded in on itself. Place the folded
    filter in a 50 mL centrifuge tube.
9.   Tighten the cap as tightly as possible. The cap will seal tightly after an additional % turn past the
    point at which initial resistance is met.
10.  Prepare a sample label (with pre-printed sample ID number sample  type "PBIO"), including the
    volume filtered, and attach it to the centrifuge tube. Cover the label completely with a strip of
    clear tape. Place the centrifuge tube into a self-sealing plastic bag.
11.  Record the sample ID number of the label and the total volume of the composite index sample
    on the Sample Collection Form. Record the volume filtered in the "Periphyton Biomass" field on
    the Sample Collection Form. Double check that the volume recorded on the collection form
    matches the total volume recorded on the sample label.
12.  Place the centrifuge tube containing the filter on dry ice.
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13.4  Data Forms and Sample Inspection
After the Site Assessment Form is completed, the Field Team Crew Leader reviews all of the data
forms and sample labels for accuracy, completeness, and legibility. The other team crew
members inspect all sample containers and package them in preparation for transport, storage,
or shipment. Refer to Appendix C for details on preparing samples for shipping.
Ensure that all required data forms for the site have been completed. Confirm that the SITE-ID,
the visit number, and date of visit are correct on all forms. On each form, verify that all
information has been recorded accurately, the recorded information is legible, and any flags are
explained in the comments section. Ensure that written comments are legible, with no
"shorthand" or abbreviations. Make sure there is no marking s in the scan code boxes. Make
sure the header information is completed on all pages of each form. After reviewing each form
initial the upper right corner of each page of the form.
Ensure that all samples are labeled, all labels are completely filled in, and each label is covered
with clear plastic tape. Compare sample label information with the information recorded on the
corresponding field data forms (e.g., the Sample Collection Form) to ensure accuracy. Make sure
that  all sample containers are properly sealed.

13.5  Launch Site Cleanup
Load the  boat on the trailer and inspect the boat, motor, and trailer for evidence of weeds and
other macrophytes. Clean the boat, motor, and trailer as completely as possible before leaving
the launch site. Inspect all nets for pieces of macrophyte or other organisms and remove as
much as possible before packing the nets for transport. Pack all equipment and supplies in the
vehicle and trailer for transport. Keep equipment and supplies organized so they can be
inventoried using the equipment and supply checklists presented in Appendix A.  Lastly, be sure
to clean up all waste material at the launch site and dispose of or transport it out of the site if a
trash can is not available.
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          14    FIELD QUALITY CONTROL
          Standardized training and data forms provide the foundation to help assure that data quality
          standards for field sampling are met. This section for field sampling and data collection are the
          primary guidelines for all cooperators and field crews. In addition, repeat sampling and field
          evaluation and assistance visits will address specific aspects of the data quality standards for the
          National Rivers and Streams Assessment.

          14.1   Revisit Sampling Overview
          Revisit sampling will provide data to make variance estimates (for measurement variation and
          index period variation) that can be used to evaluate the NRSA design for its potential to
          estimate status and detect trends in the target population of sites. A summary of the repeat
          sampling design is provided in Figure 14.1.

1
Visit 1
I
Revisit Sites (4 per State)
1
Space revisits minimum of two
weeks to one month apart
I i
Collect all
samples
In situ measures
Water chemistry
Chlorophyll-a
Feriphyton
Benthos
Enterococci
Fish
Fish plugs
Fish tissue
(at select sites)
Physical habitat
Filter Blank
Enterococci
BEFOREfiltering
other samples




Revisits = Measurement
Variation + Index period
variation




i

Visit 2
r-1
Collect all
Samples
In situ measures
Water chemistry
Chlorophyll-a
Feriphyton
Benthos
Enterococci
Fish
Fish plugs
Physical habitat









Figure 14.1 Summary of the Repeat Sampling Design

14.2  Revisit Sampling Sites
A total of 200 (approximately 10%) of the target sites visited will be revisited during the same
field season by the same field crew that initially sampled the site. Revisit samples and
measurements are taken from the same reach as the first visit. Each state has four revisit sites;
two wadeable and two non-wadeable sites. For each state these sites are:

    Wadeable Revisit sites:

    •  The two wadeable revisit sites are re samples from the NRSA 2008/09 (1-4th order). They
       are in the base sites in NRSA09, and have _RVT in the panel column. For most States,
       these are the first two sites in that category, but this is not always the case.
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    Non-Wadeable Revisit sites:

    •  The two non-wadeable revisit sites are resamples from the NRSA 2008/09 (5th and
       above order). They are in the base sites in NRSA09, and have _RVT in the panel column.
       For most States, these are the first two sites in that category, but this is not always the
       case.
If a site selected for revisit sampling is dropped, then the alternate assigned to replace it should
be revisited. The alternate site is the next site on the sampling list, in the same category (i.e.
Repeat NRSA l-4th order). If a non-wadeable site is sampled with wadeable methods, the next
non-wadeable site should be selected as the revisit site. The primary purpose of this "revisit" set
of sites is to collect temporal replicate samples to provide variance estimates for both
measurement variation and index period variation. The revisit will include the full set of
indicators and associated parameters. The time period between the initial and repeat visit to a
site is,  not less than 2 weeks and not more than one month. Label the samples Visit 2 to indicate
that they are samples from revisit sites. We will not be collecting  replicate data on whole fish
tissue.  Whole fish tissue will only be collected on the first visit.
At each revisit site, a filter blank will be collected for Enterococci during the first sampling visit
(Visit 1). The crews will filter a small amount (10 ml) of sterile buffer through 1 filter, label them
and write "blank" on the label and field form, and package and submit these samples to the lab.
The filter blanks should be run before the sample is filtered. The filter blanks should be collected
on the  first field visit (Visit 1) (Figure 14.1). Detailed description of the filter blanks is found in
Table 13.2.

14.3  Field Evaluation and Assistance Visits
A rigorous program  of field and laboratory evaluation and assistance visits has been developed
to support the National Rivers and Streams Assessment Program. These evaluation and
assistance visits are explained in detail  in the Quality Assurance Project Plan  (QAPP) for the
NRSA. The following sections will focus only on the field evaluation and assistance visits.
These visits provide a quality assurance/quality control (QA/QC) check for the uniform
evaluation of the data collection methods, and an opportunity to  conduct procedural reviews as
required minimizing data loss due to improper technique or interpretation of field procedures
and guidance. Through uniform training of field crews and review cycles conducted early in the
data collection process, sampling variability associated with specific implementation or
interpretation of the protocols will be significantly reduced. The field evaluations will be based
on the  Field Evaluation Plan and Checklists. This evaluation will be conducted for each unique
crew collecting and  contributing data under this program (EPA will make a concerted effort to
evaluate every crew, but will rely on the data review and validation process to identify
unacceptable data that will not be included in the final database).
                                                                                                 	i
14.3.1 Specifications for QC Assurance Field Assistance Visits                                   o
                                                                                                 i-
Field evaluation and assistance personnel are trained in the specific data collection methods              ^
detailed in this Field Operations Manual. A plan and checklist for field evaluation and assistance           u
visits have been developed to detail the methods and procedures. The plan and checklist are             b
included in the QAPP. Table 14.1 summarizes the plan, the checklist, and corrective action               <
procedures.                                                                                      &
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           Field
           Evaluation
           Plan
It is anticipated that evaluation and assistance visits will be conducted with each Field Crew
early in the sampling and data collection process, and that corrective actions will be conducted
in real time. If the Field Crew misses or incorrectly performs a procedure, the Evaluator will note
this on the checklist and immediately point this out so the mistake can be corrected on the spot.
The role of the Evaluator is to provide additional training and guidance so that the procedures
are being performed consistent with the Field Operations Manual, all data are recorded
correctly, and paperwork is properly completed at the site.
Table 14.1 General Information Noted During Field Evaluation

              Field Logistics Coordinator will arrange the field evaluation visit with each Field Crew,
              ideally within the first two weeks of sampling.
              The Evaluator will observe the performance of a crew through one complete  set of
              sampling activities.
              If the Crew misses or incorrectly performs a procedure, the Evaluator will note it on the
              checklist and immediately point it out so the mistake can be corrected on the spot.
              The Evaluator will review the results of the evaluation with the Field Crew before leaving
              the site, noting positive practices and problems.
              The Evaluator observes all pre-sampling activities and verifies that equipment is properly
              calibrated and in good working order, and NRSA protocols are followed.
              The Evaluator checks the sample containers to verify that they are the correct type and
              size, and checks the labels to be sure they are correctly and completely filled  out.
              The Evaluator confirms that the Field Crew has followed NRSA protocols for locating the
              site.
              The Evaluator observes the complete set of sampling activities, confirming that all
              protocols are followed.
              The Evaluator will record responses or concerns, if any, on  the Field Evaluation and
              Assistance Check List.
              If the Evaluator's findings indicate that the Field Crew is not performing the procedures
              correctly, safely, or thoroughly, the Evaluator must continue working with this Field Crew
              until certain of the Crew's ability to conduct the sampling properly so that data quality is
              not adversely affected.
              If the Evaluator finds major deficiencies in the Field Crew operations the Evaluator must
              contact a NRSA QA Project Coordinator.
           Field
           Evaluation
           Checklist
           Corrective
           Action
           Procedures
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14.4   Reporting
When the sampling operation has been completed, the Evaluator will review the results of the
evaluation with the Field Crew before leaving the site (if practicable), noting positive practices
and problems (i.e., weaknesses [might affect data quality] or deficiencies [would adversely
affect data quality]). The Evaluator will ensure that the Crew understands the findings and will
be able to perform the procedures properly in the future. The Evaluator will record responses or
concerns, if any, on the Field Evaluation and Assistance Check List. After the Evaluator
completes the Field  Evaluation and Assistance Check List, including a brief summary of findings,
all Field Crew members must read and sign off on the evaluation.
If the Evaluator's findings indicate that the Field Crew is not performing the procedures
correctly, safely, or thoroughly, the Evaluator must continue working with this Field Crew until
certain of the Crew's ability to conduct the sampling properly so that data quality is not
adversely affected. If the Evaluator finds major deficiencies in the Field Crew operations (e.g.,
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major misinterpretation of protocols, equipment or performance problems) the Evaluator must
contact the following QA official:
       Sarah Lehmann, EPA National Rivers and Streams Assessment Project QA Officer
The QA Officer will contact the Project Manager to determine the appropriate course of action.
Data records from sampling  sites previously visited by this Field Crew will be checked to
determine whether any sampling sites must be redone.
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