s>EPA
United States Environmental Protection Agency
Office of Water
Washington, DC
EPA 841-B-21-011
National Lakes Assessment 2022
Field Operations
Manual
Version 1.2, May 2022
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Version History
Version
Date
Revisions and Comments
0.0
December 2021
Internal EPA version for project QAC review and comments
0.0
February 2022
Updated introduction to be consistent with other NLA 2022 manuals
Corrected typos, formatting, numbering of tables and figures
Updated field data and tracking form names to be consistent with the
NLA App
Updated sample bottle types
Added ESA conservation measures (Section 6.4, Section 10 and
Appendix C)
1.0
February 2022
Final approved document
Corrected NLA App entry inconsistencies
Clarified whole fish sample storage and shipping (Section 7.3)
Revised ESA conservation measures (Section 6.4, Section 10 and
Appendix C)
Updated base kit list
1.1
March 2022
Added a project identifier (NLA2022) for bloomWatch reports
(Section2.2.4.5)
Clarified DO probe calibration requirements (Section 5.2.2.3)
Added direction to homogenize last integrated sample if the last pull
does not fit in container (Section 5.5.3)
Updated Figure 5-4 with attachment bridle length
Corrected shipping timeframe for T-2 samples in Table B-l
Corrected cross reference errors, formatting and typos
1.2
May 2022
Corrected atrazine preservation and shipping. Atrazine samples are to
be kept chilled until analysis (Section 5.5.3.2, Section 8.4 and Appendix B
(T-2 Frozen Batched Samples))
Added Appendix D: NLA Handpicked Sites: Resampling of the National
Eutrophication Study Lakes
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NOTICE
The intention of the National Lakes Assessment 2022 (NLA 2022) project is to provide a comprehensive
"State of the Lakes" assessment for lakes, ponds, and reservoirs across the United States. The complete
documentation of overall project management, design, methods, and standards and Quality
Assurance/Quality Control (QA/QC) measures is contained in this document and companion documents,
including:
National Lakes Assessment 2022: Quality Assurance Project Plan (QAPP) (EPA 841-B-21-009)
National Lakes Assessment 2022: Site Evaluation Guidelines (SEG) (EPA 841-B-16-008)
National Lakes Assessment 2022: Laboratory Operations Manual (LOM) (EPA 841-B-16-010)
These documents together comprise the integrated set of QAPP documents. 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), phytoplankton,
zooplankton, Enterococci, environmental DNA (eDNA), algal toxins, benthic macroinvertebrates, physical
habitat, and contaminants in fish tissue. These methods are based on both the guidelines developed and
followed in the Western Environmental Monitoring and Assessment Program (Baker, et. al., 1997),
methods employed by several key states that were involved in the planning phase of this project and
prior National Lakes Assessments. Methods described in this document are to be used specifically in
work relating to the NLA 2022. All Project Cooperators should 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. 2022. National Lakes Assessment 2022. Field Operations Manual. Version 1.2. EPA 841-B-16-011.
U.S. Environmental Protection Agency, Washington, DC.
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NTENTS
TITLE COVER PAGE
NOTICE Ill
TABLE OF CONTENTS IV
LIST OF TABLES IX
LIST OF FIGURES X
ACRONYMS/ABBREVIATIONS XI
1.0 BACKGROUND 1
1.1 Selection of Sampling Locations 1
1.2 Selection and Description of Survey Indicators 2
1.2.1 Trophic Status and Water Quality Indicators 3
1.2.1.1 Chlorophyll-a 3
1.2.1.2 Secchi Disk Transparency 3
1.2.1.3 Vertical Profile Measurements 3
1.2.1.4 Water Chemistry and Associated Measurements 3
1.2.1.5 Atrazine Pesticide Screen 3
1.2.2 Biological Indicators 3
1.2.2.1 Benthic Macroinvertebrate Assemblage 4
1.2.2.2 Zooplankton Assemblage 4
1.2.3 Physical Habitat Characterization 4
1.2.4 Human Health and Recreational Use Indicators 5
1.2.4.1 CyanoHAB Visual Observations and Real-time Reporting 5
1.2.4.2 Algal toxins (microcystins and cylindrospermopsin) 5
1.2.4.3 Fecal indicator (Enterococci) 5
1.2.4.4 Fish Fillet Contaminants Indicator 5
1.2.5 Other Indicators 6
1.2.5.1 Lake Characterizations 6
1.2.5.0 Environmental DNA 6
2.0 LOGISTICS 8
2.1 Roles and Contact Information 8
2.2 Key Information and Materials 10
2.2.1 Site Maps 10
2.2.2 Forms 11
2.2.2.1 Field Forms 11
2.2.2.2 Tracking Form 11
2.2.3 Equipment and Supplies 11
2.2.3.1 Request Form 11
2.2.3.2 Base Kit 12
2.2.3.3 Site Kit 12
^ 2.2.3.4 Whole Fish Composite Sample Kit 12
^ 2.2.3.5 Field Crew Supplied Items 12
ฃ! 2.2.4 Other Resources 12
^ 2.2.4.1 Quick Reference Guide 12
U 2.2.4.2 Site Evaluation Guidelines 13
q 2.2.4.3 Quality Assurance Project Plan 13
lli 2.2.4.4 Laboratory Operations Manual 13
m 2.2.4.5 Realtime cyanoHAB Reports 13
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3.0 DAILY FIELD ACTIVITIES SUMMARY 14
3.1 Health and Safety 14
3.1.1 General Considerations 14
3.1.1.1 Recommended Training 14
3.1.1.2 Communications 14
3.1.1.3 Personal Safety 15
3.1.1.4 Sampling Equipment 15
3.1.2 Safety Equipment 16
3.1.2.1 Safety Guidelines for Field Operations 16
3.1.3 COVID Precautions 17
3.2 Recording Data and Other Information 17
3.2.1 Field Forms 18
3.3 Sampling Scenario 19
4.0 BASE SITE ACTIVITIES 23
4.1 Pre-departure Activities 23
4.1.1 Daily Itineraries and Site Packets 23
4.1.2 Instrument Checks and Calibration 25
4.1.2.1 Depth sounding equipment 25
4.1.2.2 Multi-parameter probe Meter Performance Test 25
4.1.2.3 Global Positioning System Use and Battery Check 25
4.1.2.4 Electronic Data Capture Device Battery Check (Apple iPad) 26
4.1.3 Equipment and Supply Preparation 26
4.1.4 General Equipment and Supplies for all Activities 27
4.2 Lake Verification 27
4.2.1 Equipment and Supply List 27
4.2.2 Lake Verification at the Launch Site 28
4.2.3 Locating Index Site 29
4.3 Post Sampling Activities 30
4.3.1 Equipment Cleanup and Check for Invasive Species 30
4.3.2 Shipment of Samples and Forms 31
4.3.3 Communications 31
5.0 INDEX SITE ACTIVITIES 33
5.1 CyanoHABs Visual Assessment 33
5.2 Temperature, DO, and pH profile 33
5.2.1 Summary of Method 33
5.2.2 Equipment and Supplies 33
5.2.2.1 Multi-parameter Sonde 34
5.2.2.2 Temperature Meter Calibration 34
5.2.2.3 DO Probe Calibration 34
5.2.2.4 pH Meter Calibration 34
5.2.2.5 Conductivity Calibration 34
5.2.3 Depth Profile Procedure 34
5.3 Secchi DiskTransparency 36 i/i
5.3.1 Summary of Method 36 z
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5.3.2 Equipment and Supplies 36 \
5.3.3 Procedure for Determining Secchi Transparency 36 o
5.4 eDNA Sample Collection - Index 37
5.4.1 Summary of Method 37 ฐ
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5.4.2 Equipment and Supplies 37 ^
5.4.3 Sampling Procedure 38 <
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5.5 Water Sample Collection and Preservation 38
5.5.1 Summary of Method 38
5.5.2 Equipment and Supplies 39
5.5.3 Sampling Procedure 40
5.5.3.1 Sample Collection 41
5.5.3.2 Sample Preservation 42
5.6 Zooplankton Collection 43
5.6.1 Summary of Method 43
5.6.2 Equipment and Supplies 43
5.6.3 Sam pi in g Procedure 44
5.6.3.1 Sample Collection 44
5.6.3.2 Sample Processing 46
6.0 LITTORAL AND SHORELINE ACTIVITIES 47
6.1 Littoral eDNASample Collection 47
6.1.1 Summary of Method 47
6.1.2 Equipment and Supplies 48
6.1.3 Sampling Procedure 48
6.2 Physical Habitat Characterization 49
6.2.1 Summary of Method 49
6.2.2 Equipment and Supplies 50
6.2.3 Locating the Physical Habitat Stations and Defining the Shoreline Boundary 50
6.2.3.1 Base Site Activities 50
6.2.3.2 Littoral and Shoreline Activities 51
6.2.3.3 Shoreline and Station Location Adjustments 51
6.2.4 Establishing the Physical Habitat Plots at each station 53
6.2.4.1 Physical Habitat Plot Dimensions 53
6.2.4.2 Physical Habitat Station Layout Procedures 54
6.2.5 General Observations 55
6.2.6 Estimate Substrate Characteristics 56
6.2.7 Estimate Aquatic Macrophyte Cover 57
6.2.8 Estimate Fish Habitat Cover 57
6.2.9 Estimate the Cover and Type of Riparian and Drawdown Zone Vegetation 58
6.2.9.1 Canopy Vegetation (greater than 5 m high) 58
6.2.9.2 Understory Vegetation (5m to 0.5m high) 59
6.2.9.3 Ground Cover (lower than 0.5m high) 59
6.2.9.4 Considerations for Drawdown conditions 59
6.2.10 Record Evidence of Human influence 59
6.3 CyanoHABs Visual Assessment 61
6.4 Benthic Macroinvertebrate Sampling 61
6.4.1 Summary of Method 61
6.4.2 Equipment and Supplies 62
6.4.3 Sampling Procedure 62
6.4.3.1 Site Selection and Sample Collection 62
6.4.3.2 Sample Processing in the Field 62
h 6.5 Fecal Indicator Sample (Enterococci) 65
m 6.5.1 Summary of Method 65
z 6.5.2 Equipment and Supplies 65
3 6.5.3 Sampling Procedure 66
^ 6.5.4 Sample Processing in the Field 66
^ 7.0 LAKE WIDE FISH SAMPLE COLLECTION 67
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7.1 Summary of Method 67
7.2 Equipment and Supplies 68
7.3 Sampling Procedure 69
8.0 FINAL LAKE ACTIVITIES 72
8.1 General Lake Assessment 73
8.1.1 Lake/Catchment Site Activities and Disturbances Observed 74
8.1.2 General Lake Information 75
8.1.3 Shoreline Characteristics 76
8.1.4 Qualitative Macrophyte Survey 77
8.1.5 Waterbody Character 77
8.1.6 Qualitative Assessment of Environmental Values 77
8.1.7 CyanoHAB assessment and report 78
8.2 Filtering: Processing the Fecal Indicator (Enterococci) and Chlorophyll-/! Samples 79
8.2.1 Equipment and Supplies: Fecal Indicator 79
8.2.2 Procedures for Processing the Fecal Indicator (Enterococci) Sample 79
8.2.3 Equipment and Supplies: Chlorophyll-a 82
8.2.4 Procedures for Processing the Chlorophyll-a Samples 82
8.3 Preservation of Samples 83
8.4 Preparation of Samples for Shipping 83
8.5 Data Forms and Sample Inspection 84
8.6 Launch Site Cleanup 84
9.0 FIELD QUALITY CONTROL 85
9.1 Revisit Site 85
9.2 Field Evaluation and Assistance Visits 85
9.2.1 General Information 85
9.2.2 Preparation Activities 86
9.2.3 Field Day Activities 86
9.2.4 Post Field Day Activities 87
9.2.5 Summary 87
10.0 EPA REGIONAL AND CONTRACTED CREWS ONLY: ENDANGERED SPECIES ACT CONSERVATION MEASURES
89
10.1 Always Applicable 89
10.2 When Entering or Leaving a Site andShorline Activities 90
10.3 Index Site Activities 90
10.4 Littoral Activities 90
10.5 Lake Wide Activites: Fish Sample Collection 91
10.5.1 Sites with ESAfish possible 91
10.5.2 Sites with ESA freshwater mussles possible (indirect impacts to lake habitat-based bivalves' host
fishes) 91
11.0 LITERATURE CITED 92
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APPENDIX A: EQUIPMENT & SUPPLIES 93 ^
Base Kit 93 E
Site Kit 95 ง
Human Health Fish Sampling Kit 96 ^
Electronic Forms & Labels 97 lu
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Field Crew Supplied Equipment 97 co
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Boat Equipment List 98 h
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APPENDIX B: SHIPPING GUIDELINES 100
GeneralShipping Guidelines 100
Tracking Forms and ShipmentTypes 101
Procedure for filling out and submitting tracking via the App 102
Shipping Groups: 103
T-l - Daily Water Chemistry Samples 103
1-2 - Frozen Batched Samples 103
T-3 - Non-Chilled Batched Samples 105
1T4 - Whole Fish Composite Sample - designated sites only 106
Shipping Addresses 107
APPENDIX C: NATIVE FRESHWATER MUSSEL EXAMPLES 110
APPENDIX D: NLA 2022 HANDPICKED SITES: RESAMPLING OF THE NATIONAL EUTROPHICATION STUDY LAKES
Ill
Background Ill
Field Crews and Coordination Ill
Project and Data Quality Objectives 113
NES Indicators 113
Study Design 114
Field Methods and Procedures 115
Field Crew Training 115
Daily Field Activities 115
Field Forms and Sample Shipment 115
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ES
Table 1-1 Summary table of indicators and sampling location 7
Table 2-1 Personnel to call for specific types of questions and support needs 9
Table 2-2 Contact information 9
Table 3-1 Guidelines for recording field measurements and tracking information 18
Table 4-1 Instrument checks and calibration 26
Table 4-2 Stock solutions, uses, and methods for preparation 26
Table 4-3 Equipment and supplies - all activities 27
Table 4-4 Equipment and supplies - lake verification 27
Table 5-1 Equipment and supplies - temperature, pH, and DO profiles 33
Table 5-2 Equipment and supplies - Secchi disk transparency 36
Table 5-3 Equipment and supplies - index eDNA samples 38
Table 5-4 Equipment and supplies - water samples 39
Table 5-5 Equipment and supplies - zooplankton collection 43
Table 5-6 Lengths and numbers of zooplankton tows based on Index Site depth 45
Table 6-1. Equipment and supplies -fish littoral eDNA samples 48
Table 6-2 Equipment and supplies - physical habitat assessment 50
Table 6-3 Equipment and supplies - benthic macroinvertebrate collection 62
Table 6-4 Equipment and supplies: fecal indicator sample 65
Table 6-5 Procedure: fecal indicator (enterococci) sample collection 66
Table 7-1. Equipment & supplies: whole fish composite sample collection for human health 68
Table 7-2. Primary and secondary NLA target species for human health fish collection 71
Table 8-1 Site activities and disturbances observed during final lake assessment 74
Table 8-2 General lake information observed during final lake assessment 76
Table 8-3 Shoreline characteristics observed during final lake assessment 76
Table 8-4 Equipment and Supplies: Fecal Indicator (Enterococci) Sample 79
Table 8-5 Procedure: Processing Fecal Indicator (Enterococci) Sample 80
Table 8-6 Equipment and supplies - chlorophyll-o processing 82
Table 9-1 Equipment and supplies - field evaluation and assistance visits 86
Table 9-2 Summary of field evaluation and assistance visit information 87
Table B-0-1. Sample preservation, packaging, and holding times 108
Table D-l NES EPA Regional Crew Contact information 112 ^
Table D-2 NLA 2022 NES site water quality indicators 113 h
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Table D-3 Target number of hand-picked lakes for sampling by EPA Region 114 O
Table D-4 NES field forms 116 :~
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LIST OF FIGURES
Figure 3-1 Daily operations summary 20
Figure 3-2 Location of sample collection points and physical habitat (PHab) stations 21
Figure 4-1 Overview of base site activities 23
Figure 5-1 Secchi disk diagram (USEPA, 1991) 36
Figure 5-2 Integrated water sampler device (MPCA) 40
Figure 5-3 Procedure for using the integrated sampler device to collect depth integrated samples 41
Figure 5-4 Wisconsin net and collection bucket diagram 44
Figure 6-1 Dimensions and layout of a physical habitat station 49
Figure 6-2 Human disturbance and proximity determinations 61
Figure 6-3 D-frame net (500 pim mesh) used for collecting benthic macroinvertebrates 61
Figure 8-1 Final lake activities summary 73
Figure B-l. Sample shipping flowchart 109
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ACRONYMS/ABBREVIATIONS
ANC Acid neutralizing capacity
CPR Cardiopulmonary resuscitation
DBH Diameter at breast height
Dl Deionized
DO Dissolved oxygen
DOC Dissolved organic carbon
ESA Endangered Species Act
FLC Field Logistics Coordinator
FOM Field Operations Manual
GIS Geographic information system
GPS Global positioning system
HAB Harmful Algal Bloom
HDPE High density polyethylene
HQ Headquarters
IM Information Management
LOM Laboratory Operations Manual
MPCA Minnesota Pollution Control Agency
NARS National Aquatic Resource Surveys
NH4 Ammonium
NHD National Hydrography Dataset
NIST National Institute of Standards and Technology
NLA National Lakes Assessment
N03 Nitrate
OSHA Occupational Safety and Health Administration
PBS Phosphate buffered saline
PCBs Polychlorinated biphenyls
PDOP Position Dilution of Precision
PFAS Polyfluoroalkyl substances
PFD Personal Flotation Device
PHab Physical habitat
QA Quality assurance ^
ฆz.
QAPP Quality Assurance Project Plan O
QA/QC Quality assurance/quality control ^
QCS Quality control check solution ^
QRG Quick Reference Guide m
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SEG Site Evaluation Guidelines ^5-
SOPs Standard Operating Procedures p
TSS Total suspended solids O
QC
UL Underwriters Laboratory ^
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USGS United States Geological Survey
UTM Universal Transverse Mercator
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1.0 BACKGROUND
This manual describes field protocols and daily operations for crews to use in the National Lakes
Assessment 2022 (NLA 2022). The NLA 2022 is a statistical assessment of the condition of our nation's
lakes, ponds, and reservoirs (subsequently referred to in this manual as "lakes"). The survey is designed
to address three key questions about the quality of the nation's lakes:
1. What percent of the nation's lakes are least, moderately, and most disturbed for key indicators
of trophic state, ecological health, and human use (recreation)?
2. What is the relative importance of key stressors such as nutrients and pathogens?
3. What changes are occurring in the condition of the nation's lakes?
The surveys are also designed to help expand and enhance state and tribal monitoring programs.
Through these surveys, states and tribes have the opportunity to collect data that can be used to
supplement their existing monitoring programs or to begin development of new programs.
The NLA 2022 is one of a series of water surveys being conducted by states, tribes, the U.S.
Environmental Protection Agency (EPA), and other partners. In addition to lakes, partners will also study
coastal waters, wadeable streams, rivers, and wetlands in a revolving sequence. The purpose of these
surveys is to generate statistically-valid reports on the condition of our nation's water resources and
identify key stressors to these systems.
The NLA 2022 is designed to be completed during the summer growing season before fall lake turnover
(June through September3). Field crews will collect a variety of measurements and indicators from an
"index site" located at the deepest point of the lake up to 50 meters (or near the middle of the lake if
the lake is a reservoir), and document conditions of the littoral zone and shoreline from stations around
the lake.
1.1 Selection of Sampling Locations
EPA selected sampling locations using a probability-based survey design (Stevens and Olsen, 2004).
Sample surveys have been used in a variety of fields (e.g., election polls, monthly labor estimates, and
forest inventory analysis) to determine the status of populations or resources of interest using a
representative sample of relatively few members or sites. Using this survey design allows data from the
subset of sampled lakes to be applied to the larger target population and assessments with known
confidence bounds to be made.
With input from the states and other partners, EPA used the following framework to guide the site
selection process:
1. The National Hydrography Dataset Plus High Resolution (NHDplusHR) data layer was used to
derive a list of lakes for potential inclusion in the NLA 2022.
2. For purposes of this survey, "lakes" refers to natural and man-made freshwater lakes, ponds,
a The NLA index period is June through September. Sampling in May could be approved for lakes in areas where
stratification is expected earlier in the year. Please coordinate these requests with your Regional EPA Coordinator
and the NLATechnical Lead.
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and reservoirs greater than one hectare (approximately 2.5 acres) in the conterminous U.S.,
excluding the Great Lakes.
Mine ponds, retention basins, cooling ponds, and tidally-influenced lakes were excluded from this
study. For more information on the site exclusion criteria refer to the National Lakes Assessment
2022: Site Evaluation Guidelines (EPA 841-B-21-008).
3. The sample size was set to include 1,000 lake sampling events.
EPA used an unequal probability design to select 904 lakes and reservoirs greater than 1 hectare (ha) in
size (note: in NLA 2007, the lower size limit was 4 ha) in the continental United States. The design
includes 2 revisits in each state resulting in the target total of 1,000 site visits. Revisit samples are
collected for quality assurance purposes including evaluation of the ability of an indicator to distinguish
among sites from differences within individual sites. Of the 904 lakes, approximately 50% of the lakes
are new lakes selected for 2022 and 50% are previously sampled lakes as part of the NLA 2017. The NLA
2017 lakes are referred to as resample lakes. Also, of the 904 lakes, approximately 70% of the lakes (i.e.,
636 lakes) are designated for whole fish composite sample collection for human health. An
"oversample" list of additional lakes was also generated to allow for replacement of non-target or
otherwise unsampleable sites. The oversample list will also accommodate any state wishing to conduct a
state scale survey.
Lakes selected for the NLA 2022 are distributed among five size class categories and are spatially
distributed across the lower 48 states and nine aggregated Omernik Level 3 ecoregions (USEPA, 2013).
Related NLA 2022 documents include the following:
National Lakes Assessment 2022: Quality Assurance Project Plan (EPA 841-B-21-009)
National Lakes Assessment 2022: Site Evaluation Guidelines (EPA 841-B- 21-008)
National Lakes Assessment 2022: Laboratory Operations Manual (EPA 841- 21-010)
These documents are available at: https://www.epa.gov/national-aquatic-resource-surveys/manuals-
used-national-aquatic-resource-surveys.
1.2 Selection and Description of Survey Indicators
As part of the indicator selection process, EPA and the NLA 2022 Steering Committee evaluated
indicators used in prior NLAs, refined methodologies, and identified new indicators for NLA 2022. The
Steering Committee, comprised of state representatives from each of the EPA regions, provided advice
and recommendations to the Agency on matters related to the NLA 2022. Key screening and evaluation
criteria included indicator applicability on a national scale, the ability of an indicator to reflect various
aspects of ecological condition, and cost-effectiveness (e.g., Kurtz et al., 2001). EPA used the
Committee's recommendations to refine methods and develop final documents.
The remainder of this section briefly describes the indicators that the NLA 2022 will use to assess trophic
z status, ecological integrity, human use value, and lake characteristics (Table 1.1). Some indicators
O provide a basis for evaluating more than one category. For example, an assessment of zooplankton
ej allows for an examination of ecological integrity and trophic status, and to a certain extent, human use.
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1.2.1 Trophic Status and Water Quality Indicators
Lakes are classified according to their trophic state. "Trophic" means nutrition or growth. A eutrophic
("well-nourished") lake has high nutrients and high plant growth. An oligotrophic lake has low nutrient
concentrations and low plant growth. Mesotrophic lakes fall somewhere in between eutrophic and
oligotrophic lakes.
Chlorophyll-a, total phosphorus, and Secchi disk transparency are most often used to estimate biomass
and define the trophic state of a particular lake. Other variables are measured in conjunction with the
trophic state variables to supplement and enhance understanding of lake processes that affect primary
productivity.
1.2.1.1 Chlorophyll-a
Chlorophyll is the pigment that allows plants (including algae) to use sunlight to convert simple
molecules into organic compounds via the process of photosynthesis. Of the several kinds of chlorophyll,
chlorophyll-a is the predominant type found in green plants and algae. Measuring chlorophyll-a
concentrations in water is a surrogate for actually measuring algae biomass and it is used to estimate
trophic status.
1.2.1.2 Secchi Disk Transparency
A Secchi disk is a black and white patterned disk commonly used to measure the clarity of water based
on the distance the disk can be seen when it is lowered into the water column. The Secchi disk
measurement is used to estimate the euphotic zone depth in the field which is generally defined as two-
times the Secchi disk depth.
1.2.1.3 Vertical Profile Measurements
Depth profiles for temperature, pH, and dissolved oxygen (DO) are taken with a calibrated water quality
probe meter or multi-parameter probe sonde from the index site in each lake. This information is used
to determine the extent of stratification and the availability of the appropriate temperature range and
level of DO necessary to support aquatic life.
1.2.1.4 Water Chemistry and Associated Measurements
Water chemistry measurements are used to determine the acidic conditions, trophic state and nutrient
enrichment, and water chemistry type.
1.2.1.5 Atrazine Pesticide Screen
Atrazine pesticides are herbicides used to control the growth of weeds. Although applied to the land,
these chemicals can enter lakes via transport in water (e.g., runoff, groundwater, etc.) or atmospheric
transport. This screen will provide information about the occurrence and concentration of atrazine
pesticides in water samples from lakes across the nation.
1.2.2 Biological Indicators
Ecological integrity describes the ecological condition of a lake based on different assemblages of the
aquatic community and their physical habitat (PHab). The indicators include zooplankton, benthic
macroinvertebrates, and the physical habitat of the shoreline and littoral zones.
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1.2.2.1 Benthic Macroinvertebrate Assemblage
Benthic macroinvertebrates are bottom-dwelling animals without backbones ("invertebrates") that are
large enough to be seen with the naked eye ("macro"). Examples of macroinvertebrates include crayfish,
snails, clams, aquatic worms, leeches, and the larval and nymph stages of many insects, including
dragonflies, mosquitoes, and mayflies. Populations in the benthic assemblage respond to a wide array of
stressors in different ways so that it is often possible to determine the type of stress that has affected a
macroinvertebrate assemblage (e.g., Klemm et al., 1990). Because many macroinvertebrates have
relatively long life cycles of a year or more and are relatively immobile, the structure and function of the
macroinvertebrate assemblage is a response to exposure of present or past conditions. For the NLA, the
benthic macroinvertebrate assemblage occupying the littoral zone will be assessed, rather than the
profundal assemblage occupying the deeper regions of lakes.
Zooplankton are animal microorganisms that consist of crustaceans (e.g., copepods and cladocerans),
rotifers ("wheel-animals"), pelagic insect larvae (e.g., phantom midges), and aquatic mites. The
zooplankton assemblage constitutes an important element of the food web, where zooplankton transfer
energy from algae (primary producers) to larger invertebrate predators and fish. The zooplankton
assemblage responds to environmental stressors such as nutrient enrichment and acidification (e.g.,
Stemberger and Lazorchak 1994, Dodson et al. 2005). The effects of these environmental stressors on
zooplankton can be detected through changes in species composition, abundance, and body size
distribution.
1.2.3 Physical Habitat Characterization
The characterization of shoreline and littoral zone (the nearshore areas of a lake) physical habitat (PHab)
conditions serves three purposes. First, habitat information is essential to the interpretation of expected
lake ecological condition in the absence of human disturbance (anthropogenic impacts). Second, the
habitat evaluation is a reproducible, quantified estimate of habitat condition, serving as a benchmark
against which to compare future habitat changes that might result from anthropogenic activities. Third,
the specific selections of habitat information collected aid in the diagnosis of probable causes of
ecological degradation in lakes.
In addition to information collected in the field by the shoreline and littoral surveys, the physical habitat
description of each lake includes many map-derived variables such as lake surface area, shoreline
length, and shoreline complexity. Furthermore, an array of information, including watershed topography
and land use, supplements the physical habitat information. The shoreline and littoral characterizations
concentrate on information best derived "on the ground". As such, these results provide the linkage
between large watershed-scale influences and those influences that directly affect aquatic organisms
day to day. Together with water chemistry, the habitat measurements and observations describe the
variety of physical and chemical conditions that are necessary to support biological diversity and foster
long-term ecosystem stability. These characteristics of lakes and their shorelines are the very aspects
that are often changed as a result of anthropogenic activities.
1.2.2.2 Zooplankton Assemblage
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1.2.4 Human Health and Recreational Use Indicators
Human health indicators address the ability of the lake population to support recreational uses such as
swimming, fishing, and boating. The protection of these uses is one of the requirements of the Clean
Water Act. The NLA 2022 human health indicators include the extent of cyanobacterial harmful algal
bloom (cyanoHAB), algal toxins (microcystins and cylindrospermopsin), fecal indicator (enterococci) and
fish fillet contaminants.
1.2.4.1 CyanoHAB Visual Observations and Real-time Reporting
Cyanobacteria are microscopic organisms found naturally at low concentrations in freshwater systems.
Under optimal conditions (such as high light and calm weather, usually in summer), cyanobacteria
occasionally form harmful algal blooms (HABs), or dense aggregation of cells, that float on the surface of
the water. At higher concentrations, cyanoHAB events may be so dense that they resemble bright green
paint that has been spilled in the water. These blooms potentially affect water quality, human health
(e.g., Microcystis can produce microcystin, a liver toxin), and natural resources. Decomposition of large
blooms can lower the concentration of DO in the water, resulting in hypoxia (low oxygen) or anoxia (no
oxygen) which may result in fish kills. Field crews will be submitting real-time reports of a potential
cyanoHAB events to the state and tribal organizations that monitor water quality for recreational use
support and swimming advisories. These organizations may use this information to determine if follow
up monitoring and reporting is needed.
1.2.4.2 Algal toxins (microcystins and cylindrospermopsin)
Microcystins and cylindrospermopsin are two types of toxins produced by cyanobacteria. During a
cyanoHABs event, the toxin concentration can rapidly increase and may become elevated before a
visible bloom is observed. Elevated cyanotoxin concentrations in surface waters can persist after the
bloom fades, so human exposures can occur even after the visible signs of a bloom are gone or have
moved downstream. Exposure to elevated-levels of microcystins can potentially lead to liver damage;
the kidneys and liver appear to be the primary target organs for cylindrospermopsin toxicity.
1.2.4.3 Fecal indicator (Enterococci)
Enterococci are bacteria whose presence indicates that water may be contaminanted by human or
animal wastes. Microbes in these wastes can cause short term effects, such as diarrhea, cramps, nausea,
headaches, or other symptoms. They may pose a special health risk for infants, young children, and
people with severely compromised immune systems.
1.2.4.4 Fish Fillet Contaminants Indicator
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 for people who consume fish. Collecting whole fish composite samples and submitting
them to the laboratory for filleting and homogenization during the NLA 2022 provides sufficient tissue
for analysis of multiple chemical contaminants of concern (e.g., mercury, polychlorinated biphenyls or
PCBs, and per- and polyfluoroalkyl substances or PFAS).
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1.2.5 Other Indicators
1.2.5.1 Lake Characterizations
Observations and impressions about the lake and its surrounding catchment by field crews will be useful
for ecological value assessment, development of associations and stressor indicators, and data
verification and validation.
1.2.5.0 Environmental DNA
Two water samples will be collected and analyzed for environemtnal DNA (eDNA). Consistent with NLA
2017, one sample will be collected from the index site. The second sample will be a composite sample
from 10 littoral stations. This research measure will be used to explore indicator development for future
NLAs.
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Table 1-1 Summary table of indicators and sampling location.
Indicator
Indicator
Specifications/Location in Lake
Type
Desktop
Evaluation
Boat
Launch
Index Site
Littoral Site
Trophic and
Chemical
Indicators
Vertical profile measurements (DO,
Temperature, pH)
X
Secchi Disk transparency
X
Water chemistry (NH4, N03), major
anions and cations, alkalinity (ANC),
DOC, TSS, silica, conductivity, nutrients
(total and dissolved TN and TP)
Integrated
water sample
Chlorophyll-o
Integrated
water sample
Atrazine pesticide screen
Integrated
water sample
Biological
Benthic macroinvertebrate assemblage
10 stations
Zooplankton assemblage
(composition, structure, and size
distribution)
Vertical tow
(2 mesh sizes)
through water
column
Physical
Habitat
Physical habitat characterization
10-12
stations
Human
Health
Fecal indicator (Enterococci)
Grab
sample;
lakes
>10,000 ha
Grab sample;
last station,
lakes
< 10,000 ha
Phytoplankton (cyanobacteria)
Integrated
water sample
CyanoHAB visual observations
X
X
X
Algal toxins (microcystins and
cylindrospermopsin)
Integrated
water sample
Fish fillet contaminants
Whole fish composite sample (for fillet
analysis) collected lake wide
Other
Indicators
eDNA
1L Grab
sample;
lakes
>10,000 ha
1L Grab
sample
10 sample
composite;
1L total
Lake area, basin morphometry, and
characteristics of watershed
Using GIS
Q
Z
3
o
cc
u
<
CO
7
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2.0 LOGISTICS
2.1 Roles and Contact Information
Effective communication between field crews, EPA coordinators, and NLA 2022 contractor support staff
is essential for the survey to proceed with maximum efficiency and to ensure collection of high quality
data. This section provides:
A general description of the roles of key NLA 2022 personnel in providing logistical and technical
support to the field crews;
Flow of communication between Field Crews and these individuals (i.e., who to call for specific
types of questions or support needs); and
Contact information.
The EPA Headquarters (HQ) Project Management Team consists of the Project Leader, Logistics Leader,
and Project QA Lead, along with the EPA Technical Lead for the Fish Fillet Contaminants Indicator. The
Team is responsible for overseeing all aspects of the project and ensuring technical and QA
requirements are properly carried out. The Team is the final authority on all decisions regarding field
sampling, site evaluation, site replacement, and laboratory analysis.
The EPA Regional Coordinators are the primary EPA point of contact for Field Crews operating in their
Region. Field Crews should direct all technical and logistical questions to their EPA Regional Coordinator,
who will work with the EPA HQ Team to resolve the issue. Field Crews should also work with their EPA
Regional Coordinator to schedule an Assistance Visit to occur within the first two weeks of field
sampling. An Assistance Visit is part of the QA component of the NLA 2022 QAPP. To meet the
requirements of the QAPP, each Field Crew will allow an EPA employee or contractor to observe that
crew sampling for one day. The Assistance Visit is used to confirm the protocols are implemented as
intended and to suggest corrective actions, if needed, to the Field Crew's sampling approach.
The Information Management (IM) Coordinator provides the Field Crews with packets of forms and
labels for each site scheduled to be sampled. Crews will request these packets through a fillable PDF
Request form. The IM Team also tracks the transition of each NLA 2022 sample from the field to the
laboratory.
The Contract Field Logistics Coordinator (FLC) is responsible for tracking the Field Crew's sampling
activities and overall progress throughout the field season, ensuring that requests for supplies and
equipment are filled, and assisting Field Crews with questions concerning field logistics, equipment, and
supplies as they arise during the field season. The FLC will also review submitted status and tracking
forms to ensure that the correct samples have been taken and that those samples are being sent to the
laboratories in an appropriate timeframe.
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Table 2-1 Personnel to call for specific types of questions and support needs.
Personnel
Call
EPA Regional Coordinators
First, to ask any questions about NLA, including
questions on field protocols
Grant questions
Schedule Field Assistance Visit
EPA HQ Project Management Team
Ask questions about site access, site evaluation, and
site replacement
Ask questions about shipping locations and sample
handling procedures
Ask questions about Field Methods
Ask questions about Target Fish species
Ask questions about Survey Design
Ask questions about QA procedures
Ask questions about Laboratory Methods
If you can't reach Regional Coordinator, IM
Coordinator, or Field Logistics Coordinator
If you are unsure who to call
Personnel
ONLY Call
Information Management (IM) Coordinator
Order field forms or site kits
Submit a status report
Notify EPA about change in sampling schedule
Ask questions about submitting data packet
If EPA Regional Coordinator directs you to them
Contract Logistics Coordinator
Order replacement items for site kits, base kits, or
miscellaneous supplies
Ask questions about shipping contract, or to order
more shipping forms
If EPA Coordinator directs you to them
If you can't reach an EPA HQ or Regional Coordinator
and it is an urgent question
Table 2-2 Contact information
Title Name
Contact Information
EPA HQ Project Lead Lareina Guenzel, OW
guenzel.lareinaPepa.gov
202-566-0455
EPA HQ Project QA Lead Sarah Lehmann, OW
lehmann.sarah(ฎ eoa.gov
202-566-1379
EPA HQ Logistics Lead Brian Hasty, OW
hastv.brian(ฎ eoa.gov
202-564-2236
Contract Field Logistics Chris Turner,
Coordinator Great Lakes Environmental
Center, Inc.
cturner(ฎglec.com
715-829-3737
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Title
Name
Contact Information
EPA HQ Fish Fillet
Contaminants Indicator
Leads
Leanne Stahl, OW/OST
John Healey, OW/OST
stahl.leanne@eoa.gov
202-566-0404
Healev.iohn@eoa.gov
202-566-0176
Contract Fish Fillet
Contaminants Indicator
Trainer
Blaine Snyder
Blaine.snyderPtetratech.com
410-902-3158
Information
Management (IM)
Coordinator
Michelle Gover, GDIT
gover.michelle@epa.gov
541-754-4793
Regional EPA
Coordinators
Hilary Snook, Region 1
snook.hilarv@eoa.gov
617-918-8670
Emily Nering, Region 2
nering.emilv@eoa.gov
732-321-6764
Frank Borsuk, Region 3
Leah Ettema, Region 3
borsuk.frank@eoa.gov
304-234-0241
ettema.leah@eoa.gov
304-234-0245
Chris McArthur, Region 4
mcarthur.christooher@eoa.gov
404-562-9391
Mari Nord, Region 5
nord.mari@eoa.gov
312-886-3017
Rob Cook, Region 6
cook.robert@eoa.gov
214-665-7141
Gary Welker, Region 7
welker.garv@eoa.gov
913-551-7177
Liz Rogers, Region 8
Tom Johnson, Region 8
Rogers.liz@epa.gov
303-312-6974
Johnson.tom@epa.gov
303-312-6226
Tina Yin, Region 9
Matthew Bolt, Region 9
vin.christina@eoa.gov
415-972-3579
Bolt.matthew@eoa.gov
415-972-3578
Lil Herger, Region 10
herger.lillian@eoa.gov
206-553-1074
2.2 Key Information and Materials
2.2.1 Site Maps
Geospatial files in the form of geographic information system (GIS) design point and polygon files and
y state leaflet maps have been provided on the NLA SharePoint sites to assist in the site evaluation
process. From these files, crews should generate their own site maps with relevant information
O displayed. The site maps will be helpful in the planning and preparation for visiting and sampling a
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particular NLA 2022 site. These maps should become part of your site packet. See more information on
the site packet in Section 4.1.
2.2.2 Forms
Forms are the key to data collection and tracking for the NLA 2022. For NLA 2022 we have developed
electronic forms which will be accessed via the NLA App. Paper forms will only be provided as backups
to the App forms.
2.2.2.1 Field Forms
The NLA App is the primary way crews will record measures, observations, and collection information
during the course of the field day. Additional information regarding specifics of data entry is contained
in Section 3.2.
Electronic Field Forms: This form of data collection will be collected through an Apple iPad
which will be provided for all state, tribal, and EPA crews. Each of the field forms are separated
into sections for easier data entry. It is important for a field crew to familiarize themselves with
the NLA App prior to field sampling. Field crews should note that each individual field form must
be submitted by only one device. For example, if there are 5 field forms (A,B,C,D,E) and iPad 1
submits forms A, B, and D, then iPad 2 should not submit those 3 forms or data will be
overwritten. In this example, iPad 2 could still submit forms C and E with no issues. While a data
or Wi-Fi connection is required to submit the data, no data connection is required for the data
collection process.
Paper Field Forms: Extra paper field forms will only be provided to field crews to serve as
backup copies in case of problems with electronic field forms. As soon as possible, the
completed paper field forms should be transcribed to the NLA App for data submission. The
original completed version of the forms must also be scanned, emailed to the EPA Logistic Lead
and stored by the field crew for two years.
2.2.2.2 Tracking Form
The Tracking form in the App describes the status and location of all samples and specimens collected
during the sampling of an NLA site and is transmitted electronically to the IM Team at specified times.
When samples are shipped to the lab, a packing slip is included in the shipping container to convey to
the lab which samples are included in the shipment. These packing slips (which are pre-printed with the
same sample IDs as the individual sample labels) are included in the Label Packet with each Site Kit.
The Tracking form is divided into several shipping groups, each labelled with a shipping group number
(e.g., T-l, T-2, T-3, etc.). Sample labels, packing slips, and FedEx shipping labels also carry these shipping
group numbers to help Field Crews group correct items together for shipping. See APPENDIX B:
SHIPPING GUIDELINES for more information.
2.2.3 Equipment and Supplies
2.2.3.1 Request Form
Field Crews will submit requests for site kits, whole fish composite sample kits, and other needed
supplies via an electronic Request form provided by the FLC. This form will be submitted to the NARS IM
Coordinator who will ensure that the request reaches the appropriate entity. Crews must submit basic
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sampling information (i.e., tentative start date and number of sites crews are planning to sample) to the
FLC at or before the time of submitting request forms. Crews should submit the Request form at least
two weeks prior to their desired sampling date. The Request form will be in fillable PDF format. Users
must enter the required information, save a copy of the form to their computer or device and attach the
updated copy to an email to sampletracking@epa.gov. The IM Team will send email notification that the
request has been received.
2.2.3.2 Base Kit
The Base Kit is comprised of the subset of durable equipment and supplies needed for NLA 2022
sampling and is provided by EPA through the FLC. Typically one Base Kit is provided to each Field Crew
and contains some of the equipment that is used throughout the field season. SeeAPPENDIX A:
EQUIPMENT & SUPPLIES for a list of the items provided by EPA in the Base Kit. EPA anticipates that Base
Kitequipment will be available for use in future NLA efforts.
2.2.3.3 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 EPA through the FLC. The site kit will contain all the sample bottles
necessary for sampling a single lake. A new Site Kit should be requested for each site sampled, and
crews should consider having at least one additional site kit available as a spare should any supplies be
lost See APPENDIX A: EQUIPMENT & SUPPLIES for the consumable items that will be provided by EPA.
2.2.3.4 Whole Fish Composite Sample Kit
A sampling kit for the whole fish composite sample will be provided for all designated fish sampling
sites. In the survey design, "FT" in the "Panel_Use" identifies lakes designated for fish sampling. This
sampling kit contains consumable supplies provided by EPA. Field crews should request a new whole fish
sampling kit for each designated fish site to be sampled. See APPENDIX A: Equipment & Supplies for the
consumable items that will be provided by EPA.
2.2.3.5 Field Crew Supplied Items
The field crew will also supply particular items for the field sampling day. These items might include
supplies from the NLA 2007, NLA 2012, or NLA 2017, typical field equipment (like a global position
system (GPS) receiver or multi-paramter probe), or boat equipment. See APPENDIX A: EQUIPMENT &
SUPPLIES for the items that the field crew will need to provide.
2.2.4 Other Resources
The complete documentation of overall project management, design, methods and standards, and
QA/QC measures is contained in this document and companion documents (listed in NOTICE and
described below). The NLA 2022 participants must agree to follow the QAPP, including the protocols and
design, and the associated documents - the NLA 2022 FOM, LOM, and SEG.
2.2.4.1 Quick Reference Guide
Field crews will NOT receive a NLA 2022 QRG. All components of the QRG will be contained in the FOM
and as "i" buttons in the NLA App. The electronic version of the FOM will be provided on the EPA issued
Apple iPads and will be a searchable document. Detailed steps to complete protocols can be found in
the FOM. Many of the tables and figures in the FOM will also be included in the App.
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2.2.4.2 Site Evaluation Guidelines
The NLA 2022 SEG (EPA 841-B-16-001) outlines the process to compile the final list of candidate lakes for
sampling. The process includes locating a candidate lake, evaluating the lake to determine if it meets the
criteria for inclusion in the target population and is accessible for sampling, and if not, replacing it with
an alternate candidate lake.
2.2.4.3 Quality Assurance Project Plan
Large-scale and/or long-term monitoring programs such as those envisioned for national surveys and
assessments require a rigorous QA program that can be implemented consistently by all participants
throughout the duration of the monitoring period. QA 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 NLA 2022 QAPP (EPA 841-B-16-003) and the field crew leader is required to sign
the QAPP signature page prior to beginning field sampling activities. The QAPP contains more detailed
information regarding 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 project level QA procedures, refer to the NLA 2022 QAPP.
2.2.4.4 Laboratory Operations Manual
The methods used for the laboratory sample analysis is available in the NLA 2022 Laboratory Operations
Manual (LOM) (EPA 841-B-21-010).
2.2.4.5 Realtime cyanoHAB Reports
Field crew leads are to report a cyanoHAB event if a bloom is occuring at the time of sampling. The
Apple iPads are preloaded with the bloomWatch application to support realtime reporting of a bloom to
the appropriate state, tribal and/or watershed organization authority. Alternatively, field crews are
encouraged to download state-specific monitroing and reporting tools that will be in use for the 2022
recreation season. Additional information on state-specific HABs monitoring programs can be found
here (https://www.epa.gov/cyanohabs/state-habs-monitoring-programs-and-resources). To submit a
report to bloomWatch or a state-specific tool, the field crews should should be prepared with emails for
state officals that are to receive these report (i.e., officals that monitor and report on cyanobacterial
bloom events). Field crews must select the preferred reporting approach and be familiar with its input
needs (e.g., extent, photos, email contacts etc.) prior to initiating field work. All reports submitted via
the bloomWatch app should add the project identifier 'NLA2022' in the comment section in the app.
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3.0 DAILY FIELD ACTIVITIES SUMMARY
This section presents a general overview of the activities that a field crew conducts during a typical 1-
day sampling visit to a lake. The following sections include general guidelines for health and safety,
recording data, and using standardized field data forms and sample labels.
3.1 Health and Safety
Collection and analysis of samples can involve risks to personal safety and health, and the safety of the
field crew must always be the primary consideration during sampling. This section describes general
safety considerations, some safety equipment, and safety guidelines for field operations.
This section does not substitute for an official Health and Safety Plan. The crew MUST ALWAYS
carefully follow the protocols in their Health and Safety Plan for the NLA field work that was approved
by the state, tribe, or other organization with which the field crew is affiliated. The crew should carry
a copy of this approved Health and Safety plan in the field.
3.1.1 General Considerations
It is the responsibility of the group safety officer or project leader to ensure that the necessary safety
courses are taken by all field personnel and that all safety policies and procedures are followed. Each
state, tribe, or other organization must have a specific safety plan for the sampling the NLA sites,
including a communications plan that addresses safety and emergency situations. The plan should have
a daily check-in procedure for field personnel, and emergency contacts for police, ambulance, fire
departments, hospitals, and search and rescue personnel. Important considerations related to field
safety are listed below. It is the responsibility of the group safety officer or project leader to ensure that
the necessary safety courses are taken by all field personnel and that all safety policies and procedures
are followed. Sources of information regarding health and safety considerations and/or safety-related
training include the American Red Cross (http://www.redcross.org/rn/phssmrd/take-a-class), the
National Institute for Occupational Safety and Health (1981) (see the most recent revisions at
https://www.cdc.gov/niosh/docs/81-123/, and the U.S. Coast Guard
(http://www.uscgboating.org/recreational-boaters/boating-safety-courses.php).
First aid;
Cardiopulmonary resuscitation (CPR);
Vehicle safety (e.g., operation of 4-wheel drive vehicles, trailer towing and maneuvering);
Boating and water safety;
Field safety (weather, personal safety, orienteering, site reconnaissance prior to sampling);
Equipment design, operation, and maintenance; and
Handling of chemicals and other hazardous materials.
3.1.1.2 Communications
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
3.1.1.1 Recommended Training
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with their supervisor for safety. An emergency communications plan should include contacts for police,
ambulance, fire departments, hospitals, and search and rescue personnel. Below are some items to
address:
Check-in schedule;
Sampling itinerary (vehicle used & description, time of departure & return, and travel route);
Contacts for police, ambulance, hospitals, fire departments, and search and rescue personnel;
Emergency services available near each sampling site and base location; and
Cell (or satellite) phone number, if possible.
3.1.1.3 Personal Safety
Proper field clothing should be worn to prevent hypothermia, heat exhaustion, sunstroke, drowning, or
other dangers. Field personnel should be able to swim, and a personal flotation device (PFD) must be
used. Chest waders made of rubberized or neoprene material and suitable footwear must always be
worn with a belt to prevent them from filling with water in case of a fall. Below are some personal safety
items to address:
Field clothing and other protective gear including lifejackets for all crew members;
Medical and personal information (allergies, personal health conditions, and required
medications);
Personal contacts (family, telephone numbers, etc.); and
Physical exams and immunizations.
Many hazards lie out of sight in the bottoms of lakes, 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.
Lakes and surrounding landscapes can be home to dangerous organisms. Field crews should take care to
minimize contact with biting insects, bees, poisonous snakes and dangerous animals. Insect repellent
and protective clothing will help to limit exposure. At the end of each field day, workers should inspect
their bodies for ticks. Any person allergic to bee stings, other insect bites, or plants (i.e., poison ivy,
poison oak, poison sumac, etc.) should take proper precautions and have any needed medications on
hand. In addition, field crew members should always be aware of their surroundings to protect
themselves from dangerous animals, such as alligators, mountain lions, bears, and wolves.
3.1.1.4 Sampling Equipment
Field crew members should be familiar with hazards associated with the use of sampling equipment and
establish appropriate safety practices prior to their use. They must ensure that all equipment is in safe
working condition.
Because boats are used to access NLA sampling sites, personnel must be trained in operating the type of
boat in use incouding appropriate state or other certifications. Personnel must consider and prepare for
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hazards associated with the operation of motor vehicles, boats, winches, tools, and other incidental
equipment. Boat operators should 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 and online (U.S. Coast Guard,
https://www.uscgboating.org/images/420.PDF ). All boats with motors must have fire extinguishers,
boat horns, life jackets or flotation cushions, and flares or communication devices.
3.1.2 Safety Equipment
Appropriate safety apparel such as life jackets, waders, 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 for use in case of an emergency. Supplies such as anti-bacterial soap and an adequate
supply of clean water or ethyl alcohol must be available for cleaning exposed body parts that may have
been contaminated by pollutants in the water or sediments.
3.1.2.1 Safety Guidelines for Field Operations
General safety guidelines for field operations are presented below.
Personnel participating in field activities on a regular or infrequent basis should be in sound physical
condition and have a physical examination annually or in accordance with Regional, State, or
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 standardized health and hazards procedures (e.g., National Institute for Occupational
Safety and Health [1981]; for the most recent revisions see https://www.cdc.gov/niosh/docs/81-123/;.)
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 any 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, may incur
personal liability or liability for the crew members and their respective organizations, may cause
personal injury, or may cause unbudgeted expenditure of time and money for proper treatment
and disposal of the material. However, it is important not to ignore environmental incidents.
Notify the proper local, state, and/or federal authorities of any incident of this type so that they
may take the necessary actions to properly respond to the incident.
3. For most environmental incidents, the following emergency telephone numbers should be
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provided to all field crews: State or Tribal department of environmental quality or protection,
U.S. Coast Guard, and the 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.
Specific Safety Guidelines are below:
Two persons must be present during all sample collection activities, and no one should be left
alone while in the field.
Minimize exposure to lake water and sediments as much as possible. Use gloves when
necessary, and clean exposed body parts as soon as possible after contact.
All electrical equipment must bear the approval seal of Underwriters Laboratories (UL) 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, specialized garments, etc.)
when handling and using hazardous chemicals.
Persons working in areas where venomous snakes may be encountered must check with the
local Drug and Poison Control Center for recommendations on what should be done in case of a
bite from a venomous snake.
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 (e.g., an "Epi-Pen").
Protect yourself against the bite of deer or wood ticks because of the potential risk of acquiring
pathogens that can cause Rocky Mountain spotted fever, Lyme disease, and other diseases.
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 10ฐC or 50ฐF) if he or
she is exposed to wind or becomes wet.
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 of any chemicals in the field.
3.1.3 COVID Precautions
Safety is the number one concern for all personnel. In implementing the NLA, crews should follow their
agencies' guidance on maintaining social distance, use of personal protective equipment, travel
restrictions, sanitizing equipment, vehicles and boats, and if necessary, hotel rooms. NLA training and
assistance visits will be implemented in a manner that considers Covid-19 safety requirements and
restrictions.
3.2 Recording Data and Other Information
All samples need to be identified and tracked; and associated information for each sample must be
recorded. 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 lake if the initial information recorded was inaccurate. As mentioned in Section 2.2.2, there
are two forms for collecting sample data: electronic field forms and backup paper field forms. See below
for important information pertaining to data entry for each of these forms.
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3.2.1 Field Forms
The NLA 2022 field data and tracking forms are formatted in the NLA App so that the data you record
can be electronically submitted to the NARS IM database. It is important that field data and sample
information are recorded accurately and consistently. General guidelines for recording field
measurements are presented in Table 3-1. More detailed instructions for filling out specific forms are
provided in each protocol chapter of this manual.
Table 3-1 Guidelines for recording field measurements and tracking information.
ACTIVITY GUIDELINES
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Field Measurements
Data Recording
Record observations and measurement values only using the official NLA App (provided
on EPA owned Apple iPads for all regional, state, and tribal crews).
If you make an error when recording data and changes are required, it is best to enter
the new value and resubmit that electronic form.
Use the correct crew ID assigned during field training.
Use the units and formats specified on individual data forms for recording data.
For any sample or data where additional explanation is needed, use the provided
comment bubble adjacent to the data
Sample Collection
Sample Labels and
Tags
Use a writing instrument that leaves clear, dark text to record information (e.g., a No. 2
pencil on paper tags or a water or smear proof fine-point indelible marker on adhesive
labels). Use the sample-type appropriate adhesive labels with preprinted Sample ID
numbers for each sample. Be sure to fill in any requested information about the
sample on the sample label and affix it to the outside of the sample container. Cover
completed labels with clear tape.
Place a waterproof paper tag inside each benthic macroinvertebrate collection jar with
the required information written with a No. 2 lead pencil.
Sample Collection
Information
Record that each sample has been collected on the appropriate data form. Be sure to
cross-check the Sample ID number from labels and tags with the Sample IDs populated
in the Tracking form .
QA and Tracking
Before Leaving Site:
Review of Data
Forms and
Comparison of
Sample Labels and
Data Forms
Review all data forms for accuracy and completeness.
Review all sample labels for accuracy, completeness, and legibility.
Verify that the information recorded on the sample labels and tags is consistent with
all Sample IDs listed on the Tracking form in the NLA App.
Before Shipping
Samples: Review of
Sample Labels and
Tracking Form
Complete all sections in the Tracking form required for all samples being shipped.
Review the Tracking form for consistency and correctness.
Compare labels on samples with the Sample IDs recorded on the Tracking form for
accuracy and completeness before shipping samples.
Review of Data
Forms
The Field Crew Leader should review the completed forms in the NLA App as soon as
is practicable to ensure they are complete and all data forms are consistent and
correct
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ACTIVITY GUIDELINES
Confirm that the forms have been reviewed by selecting the reviewed bubble in the
App for each electronic data form.
If any revisions are made, re-submit the updated form(s) as soon as possible to
update the IM Database.
After each submission, a data summary email will be sent to the email address which
submitted the data. This data summary contains a list of the data forms and their
most recent submissions date/time as well as a list of the most critical data points
collected at the site. The Field Crew Leader should review this data summary to
ensure that the data forms were successfully received and that critical values are
present and correct.
3.3 Sampling Scenario
Field methods for the NLA 2022 are designed to be completed in one field day for most lakes.
Depending on the time needed for both the sampling and traveling for that day, an additional day may
be needed for pre-departure and post-sampling activities (e.g., cleaning equipment, repairing gear,
shipping samples, and traveling to the next lake). Remote lakes with lengthy or difficult approaches may
require more time to gain access to the lake, and field crews will need to plan accordingly.
A field crew typically will consist of at least two people. Two people are always required in the boat
together to execute the sampling activities and to ensure safety. Any additional crew members may
either remain on shore to provide logistical support or be deployed in a second boat to assist in data
collection. Figure 3-1 and Figure 3-2 present a daily field sampling scenario showing how the workload
may be split between crew members. Each field crew should define roles and responsibilities for each
crew member to organize field activities efficiently. Minor modifications to the sampling scenario may
be made by crews; however, the sequence of sampling events presented in Figure 3-1 cannot be
changed and is based on the need to protect some types of samples from potential contamination and
to minimize the possibility of holding time exceedance once samples are collected. The following
sections further define the sampling sequence and the protocols for sampling activities.
NOTE: When sampling large lakes (lakes > 10,000 hectares), field crews may omit the physical habitat,
benthic macroinvertebrate, and littoral eDNA sampling efforts altogether.
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Collect whole fish composite sample at designation fish tissue (FT) sites |
HB Timing and location of collection at crew's discretion j
Locate and travel to physical habitat stations
Documentvisualobservationsof cyanoHAB events at habitat stations Collect littoral composite fish eDNA sample from surface water
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Conduct habitat characterisations
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Sample benthlc macroinvertebrates
i
Col I ect fecal i nd ic ator sam pie at I ast h abitat stati on
f
-
Return to launch site
-
Filter Enterococrisample
Muat be filtered and frozen within 6 hours of collection
Fi Iter ch torophyl l-a sam pi e
Preserve benthic, zoopJankton, and phytoplankton sample
Check and prepare all samplesfor transport or shipment
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Clean and organizeequipmentfor loading
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Inspect and clean boat, motor, and trailerto preventtransfer of
nuisance species and contaminants
Revi ew App d ata forms for com pi eten ess
J
Su bm it d ata vi a th e App /Ship sam pies/Su bm it App Tracki ng
Figure 3-1 Daily operations summary
20
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Profunda I Zone
10 evenly
spaced
sampling
stations
FINAL HABITAT
STATION
Feca! indicator sample
Observation Station
LAKEWIDE FISH COLLECTION
(fish habitat throughout the
lake)
Riparian
Plot
> 15 m
Benthik: sample collected
from dominant habitat within
littoral zone
10 m
Littoral Zone
Sub-Littoral Zone
Shoreline
band (1m)
INDEX SITE
Temp, DO, pH profile, in situ
FisheDNA
15 m
A
Drawdown
Variable or
nonexistant
HABITAT STATIONS (A-J)
Phryical Habitat, Benthic, & Littoral eDNA
sampling, and cyanoHAB observations
s taning point randomly selected a priori
Water Chemistry
Chlcrophyll-a
Phytoplankton
Atrazine
AigalToxins
Zoopiankton
cyanoHAB observations
Figure 3-2 Location of sample collection points and physical habitat (PHab) stations.
The field crew arrives at the lake in the early morning to complete the sampling in a single day. The eg
sampling sequence is to: 2
1. verify lake, calibrate equipment, locate, and travel to the index site; ->
in
2. document visual observations of a cyanoHAB event at boat launch and index site; ^
3. conduct depth profile measurements of DO, temperature, and pH; t
4. take Secchi disk transparency depth measurement; h
5. collect fish eDNA sample; ^
6. use the integrated sampler to collect water chemistry, chlorophyll-o, atrazine, algal toxin, and yj
LL.
phytoplankton samples; >;
7. collect zoopiankton samples; <
21
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8. collect fish eDNA sample for composite sample from the littoral plot at the ten PHAb stations
(A,B,C,D,E,F,G,H,I,J) prior to initiating physical habitat characterization or benthic sampling;
9. document visual observations of cyanoHABs and conduct physical habitat characterization
around the margin of the lake at ten PHab stations (A,B,C,D,E,F,G,H,I,J);
10. collect benthic samples at ten PHAb stations (A,B,C,D,E,F,G,H,I,J) concurrent with physical
habitat characterization;
11. collect whole fish composite samples;
12. collect fecal indicator sample at last PHab station (<10,000 ha lakes) or boat launch when exiting
lake (>10,000 ha lakes);
13. filter enterococci and chlorophyll-o samples;
14. preserve and prepare all samples for shipment;
15. review App field forms ;
16. report sampling event; and
17. ship time-sensitive samples (water chemistry, chlorophyll-o, eDNA, and fecal indicator samples).
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4.0 BASE SITE ACTIVITIES
Field crews are to 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 lake site and
after return from the field (Figure 4-1). Close attention to these activities is required to ensure that the
field crews know:
where they are going;
that access is permissible and possible;
that equipment and supplies are available and in good working order to complete the sampling
effort; and
that samples are packed and shipped appropriately.
PREDEPARTURE ACTIVITIES
Crew Leader Crew Members
Prepare daily itinerary Instrument checks & calibration
Equipment & supplies preparation
Whole Crew
Lake Verification
POST SAMPLING ACTIVITIES
Crew Leader Crew Members
Review forms & labels Clean boats with 1% bleach solution and perform
Package & ship samples & data forms safety checks (boat, trailer, & equipment)
File Submit status report with regional coordinatorform Charge or replace batteries
Refuel vehicle and boat
Obtain ice and other consumable supplies as needed
Figure 4-1 Overview of base site activities
4.1 Pre-departure Activities
Pre-departure activities include developing daily itineraries, checking and calibrating instruments, and
preparing equipment and supplies. Procedures for these activities, which will take place at your office or
laboratory, camping site, or motel, are described in the following sections.
4.1.1 Daily Itineraries and Site Packets
The Field Crew Leader is responsible for developing daily itineraries and a site packet. A site packet
contains information key to the planning and preparation for visiting and sampling a particular NLA site.
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Development of site packets should have been initiated during site evaluation and reconnaissance (See
NLA 2022 Site Evaluation Guidelines); however, the field crew may need to gather additional
information for the site packet during preparation for the sampling visit. Also, it is the responsibility of
the field crew to obtain access permissions and any needed permits as part of developing the site
packet.
Prior to a field crew traveling to a NLA site for sampling, the information for the site packet must be
gathered and reviewed. Site packet development entails compiling maps, contact information, copies of
permission letters, and access instructions. The Field Crew Leader must be sure to lay out the physical
habitat (PHab) stations on a site map before the sampling day (see Section 6.2.3). Additional activities
include confirming the best access routes, calling the landowners or local contacts, and confirming
lodging plans. The site packet may include the following documents:
Site maps: Generated by crew (see Section 2.2.1 Site Maps).
Other Maps, Imagery, or GIS Data: Any other maps, aerial photos, GIS data, or sources of
information compiled by Field Crews and/or their partners that could be helpful to sampling the
NLA sites.
Land Ownership Status, Requirements and Permissions for Access:
o Landowner identity and contact information,
o Results of communication with landowners,
o Documentation of permission to access private land.
o Permissions for crossing private lands to reach sites located on public lands,
o For public land, response of relevant agency to notification that you will be accessing a
site, and, if needed, permissions to do so.
Permits: Any permits or documentation required for site access, or for data collection activities
or sample/specimen collection.
Information for Accessing the Site:
o Contact information for landowners.
o Notes about whether landowner(s) want to be informed when Field Crew is on site,
o Contact information for individuals who must be available to open gates or allow entry
to a site, and the time and location for meeting them,
o Notes on locked gates, pets, livestock, or other things that could impede access,
o Notes about active hunting, farming, mining, or other activities on or near the site,
o Current conditions that could prevent access (e.g., high water, forest fires, road
closures, etc.).
Site Evaluation Notes:
o Site Evaluation notes, annotated aerial photos, sketch map, and completed site
evaluation form that can aid with planning for accessing or sampling a site.
Driving and Hiking Routes to the Site:
o Detailed driving directions may be obtained from the NLA Google Earth files,
o Results from the Site Evaluation may include driving directions and notations about site
access or logistically challenging conditions on the site, which can be useful in relocating
the site or helpful in anticipating special circumstances.
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Preliminary Plan for Establishing Physical Habitat Stations: As part of the base location
activities to prepare for field work, review aerial photos and maps of the site and make a plan
for laying out the PHab stations. This plan should be included in the site packet.
Federally Protected Species (EPA Regional and Contract Crews ONLY): See Section 10.0.
Any other site specific information (e.g., road construction and road closings) useful to the Field
Crew.
4.1.2 Instrument Checks and Calibration
Test and calibrate instruments prior to sampling. You can calibrate instruments and probes prior to
departure for the lake site or at the lake, with the exception of the DO probe. Because of the potential
influence of altitude, calibrate the DO probe at the lake site (NOTE: some newer instruments may allow
for calibration independent of altitude). Field instruments include a multi-parameter probe unit for
measuring temperature, DO, and pH and a GPS receiver. Field crews should have access to backup
instruments if any instruments fail the manufacturer performance tests or calibrations.
4.1.2.1 Depth sounding equipment
Crews are responsible for checking the accuracy of their devices against a sounding line at the beginning
of the field season to verify that the device is providing accurate depth information.
4.1.2.2 Multi-parameter probe Meter Performance Test
Test and pre-calibrate the multi-parameter probe meter prior to departure from the base site, following
either the Standard Operating Procedure (SOP) developed for the instrument or the manufacturer's
calibration and maintenance procedures. Field crews should perform a QC check of the pH meter
calibration (and conductivity meter calibration, if this optional measurement is taken) at regular
intervals designated by the manufacturer. Field crews will be responsible for preparing or purchasing
their own QC solution.
4.1.2.3 Global Positioning System Use and Battery Check
A GPS unit is used to locate the launch, index site and each of the physical habitat stations. Therefore, it
is imperative that the Field Crew understands how to operate their GPS unit. The Global Positioning
System (GPS) uses signals sent from multiple orbiting satellites to a ground-based sensor in order to fix a
position on the earth. GPS uses signals sent from multiple orbiting satellites to a ground-based sensor in
order to fix a position on the earth. Position accuracy depends on the Position Dilution of Precision
(PDOP) which is a measure of the geometry of the satellite spread over the location of the observer. Low
PDOP values are typically conveyed to the user as a measure of accuracy or precision and represent
more advantageous satellite geometry and therefore less locational error. For NLA, crews should
regularly monitor the accuracy reading on their GPS and should record coordinates only after achieving
the lowest amount of error possible.
GPS uses many alternative mathematical models to describe the spherical shape of the earth and each is
a separate datum. Commonly used datums include NAD27 CONUS, NAD83, and WGS84. Each represents
a different interpretation of the shape of the earth. The NARS standard is NAD83. Thus, all GPS units
should be switched to this standard - prior to completing any field activities. Crews should also confirm
that the NAD83 datum is being used when the GPS is turned on prior to data collection. If the GPS is not
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set for NAD83 and the unit cannot be changed readily, note the datum used on the data forms for later
conversion.
GPS devices use a variety of units for position designation based on an imaginary latitude and longitude
coordinate grid system laid across the earth (degrees, minutes, seconds, or degrees and decimal
minutes, and UTM (a metric system). The NARS standard is decimal degrees for reporting all GPS
positions.
Refer to the GPS user's manual to provide specific instructions on setting the Datum, coordinate system,
and units to NLA standards. Turn on the GPS receiver and check the batteries prior to departure.
Replace batteries immediately if a battery warning is displayed.
4.1.2.4 Electronic Data Capture Device Battery Check (Apple iPad)
Charge the Apple iPad each night and check the power prior to departure. External battery packs are
often available for these devices if battery life is a concern.
Table 4-1 Instrument checks and calibration
Equipment Preparation
GPS Unit
Check the batteries prior to departure
Ensure map datum is set to NAD83
Ensure locational units are set to decimal degrees
latitude xx.xxxxxx; longitude -xxx.xxxxxx
Perform manufacturer checks as necessary to ensure accuracy
Multi-parameter Probe
Calibrate per manufacturer guidelines (DO to be calibrated at lake)
Check the batteries prior to departure
Perform QC Check as directed by manufacturer and/or laboratory protocols (field
crews will supply QC check solution)
Electronic Data Capture
Device (Apple iPad)
Check the power prior to departure
Ensure NLA Data collection App is installed, up to date, and functioning
4.1.3 Equipment and Supply Preparation
Check your inventory of supplies and equipment prior to departure using the equipment and supplies
checklists provided in the Appendix; use of the lists is strongly recommended. Pack meters, probes, and
sampling gear in such a way as to minimize physical shock and vibration during transport. If necessary,
prepare stock preservation solutions as described in Table 4-2. Follow the regulations of the
Occupational Safety and Health Administration (OSHA).
Table 4-2 Stock solutions, uses, and methods for preparation.
Solution Use Preparation
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Bleach (1%)
Clean nets, other gear, and inside of
boat.
Add 40 mL bleach to 4,000 mL distilled water.
Lugol's
Preservative for phytoplankton
samples.
Lugol's will be supplied with base kit.
If preparation is needed: Dissolve 100 g Kl in 1 L of
distilled water. Dissolve 50 g iodine (crystalline) in
100 mL glacial acetic acid. Mix these two solutions.
Remove any precipitates. Store in the dark.
95% Ethanol
Preservative for benthic invertebrate
samples and zooplankton samples.
No preparation needed (use stock solution as is).
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Refuel vehicle(s) and conduct maintenance activities the night before a sampling trip. Check trailer
lights, turn signals, and brake lights before departure. In addition, inspect your vehicles, boats, and
trailers every morning before departure. Pay particular attention to the trailer hitch, electrical
connections, tie downs, tire pressure, and the overall condition of the boats.
Label and package as many of the sample containers as possible in the site kit prior to departure.
Container labels should not be covered with clear tape until all information is completed during
sampling at the lake. Store an extra kit of sampling supplies (Cubitainersฎ, bottles, filters, foil, gloves,
forms, pencils, permanent markers, and labels) in the vehicles. Inventory these extra supply kits prior to
each lake visit. Be sure to order field sampling site kits well in advance (two week minimum) by
submitting the electronic Request form.
4.1.4 General Equipment and Supplies for all Activities
Table 4-3 indicates equipment and supplies that will be used for all activities.
Table 4-3 Equipment and supplies - all activities.
Type
Item
Quantity
Forms
NLA App pre-installed on Apple iPad
2
Reference
NLA 2022 Field Operations Manual (FOM)
NLA 2022 Quality Assurance Project Plan (QAPP)
NLA 2022 Site Evaluation Guidelines (SEG)
NLA 2022 Fact Sheets
1
1
1
10
Documentation
Clipboard
Pencils (#2, for benthos inner sample tags)
Permanent markers (fine tip, for adhesive labels)
Labels
Field Notebook (optional)
Clear tape strips (to cover sample labels)
1
1
1
1
As needed
Collection
Access permission documents/permit (if required)
1
4.2 Lake Verification
4.2.1 Equipment and Supply List
Table 4-4 is the checklist for equipment and supplies required to conduct protocols described in this
section. It is similar to, but may be somewhat different from, the checklist that is used at a base site to
assure that all equipment and supplies are taken to, and available at, the lake. Field crews should use
the checklist presented in this section to ensure that the equipment and supplies are organized and
available on the boat in order to conduct protocols correctly and efficiently.
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Table 4-4 Equipment and supplies - lake verification.
Type
Item
Quantity
Form
NLA Verification Form
1
Collection
Depth Finder (hand-held or boat mounted sonar)
1
GPS unit (with manual, reference card, extra battery)
1
Anchor (with 75 m line or sufficient to anchor in 50 m depth)
1-2
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4.2.2 Lake Verification at the Launch Site
Before sampling activities begin, you must verify that you are at the correct lake and whether it meets
the criteria for sampling. Confirming that you are at the correct lake is based on map coordinates,
locational data from the GPS when possible, and any other evidence such as signs or conversations with
local residents. The Design Name for the site is pre-populated in the App (UNKNOWN if no name), crews
should populate the Lake Name if known. Record locational coordinates for the lake on the Verification
form. If GPS coordinates are obtained, fill in the bubble to indicate GPS and record the latitude,
longitude in decimal degrees.
Determine the number of satellites being used by the GPS and select if 3 or fewer satellites or if 4 or
more satellites were used. All coordinates will be recorded in the NAD83 datum. Compare the map
coordinates pre-populated in the App for the lake with the GPS coordinates you record and verify that
you are at the correct lake. [Note: The map coordinates in the App represent the "labeling point" in NHD
and may not be near either the index site or the launch site]. Verification that you are at the target lake
can be confirmed via other methods (e.g., map, landowner confirmation). If GPS coordinates are not
available, do not record any information, but try to obtain the information at a later time during the
visit. A GPS location may be taken at any time during a lake visit and recorded by flagging the launch site
coordinates and providing a comment.
Record directions to the lake and a description of the launch site on the Verification form regardless of
whether the site is sampled or not. This information is very important and will be used in the future if
the lake is revisited by another sampling crew. Provide information about signs, road numbers, gates,
landmarks, and any additional information you feel will be useful to another sampling crew in locating
this lake in the future. It is also helpful to describe the distance traveled (miles) between turns. Also
describe the launch site. For example: Can the boat be launched with a trailer? Are there fees? Is the
launch paved or does it consist of soft sand? What landmarks are at the launch? Due to privacy
concerns, do not record landowner contact information (e.g., name, address, phone, email address) on
the Verification form.
In addition to, or in the absence of, an accurate GPS reading, use as many of the following methods as
possible to verify the site:
Obtain confirmation from a local person familiar with the area.
Identify confirming roads and signs.
Compare the lake shape to that shown on a topographic map (United States Geological Survey
(USGS) 7.5-minute map or equivalent).
Determine lake position relative to identifiable topographic features shown on the map.
If the lake shape on the USGS topographic map does not correspond with the actual lake shape from
your site map, and you cannot verify the lake by any other means, check "Not Verified" and provide
comments on the Verification form. At each lake, evaluate whether or not the lake meets the NLA
operational definition of a "lake":
> one ha in total surface area;
> 1,000 square meters of open water;
> one meter in depth;
Not saline (due to saltwater intrusion or tidal influence); and
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Not used for aquaculture, disposal-tailings, mine-tailings, sewage treatment, evaporation, or
other unspecified disposal use.
If the lake does not meet this definition, select the "non-target" bubble in the lake sampled section on
the Verification form and provide an explanation for not sampling the lake. Add any additional
explanation as required. (For complete details on the Site Evaluation process, refer to the companion
document Site Evaluation Guidelines [EPA 841-B-06-003]).
Record the names of each crew member on the Verification form. At the launch site, document visual
observations of a potential cyanobacterial bloom in the NLA App and photo document the bloom
according to the reporting mechanism selected by the field crew (i.e., bloomWatch or state-specific
reporting mechanism).
Regardless of whether the lake is sampled or not, the field crew must fill out and submit a Verification
form for every lake that is visited with the intent to sample.
4.2.3 Locating Index Site
When determining lake origin, i.e., lake vs. reservoir in the field, a body of water that was a stream or
river and subsequently dammed to create a lake is considered a reservoir. A lake which has had its level
raised because of a dam is an "enhanced" lake and will be considered a natural lake for NLA 2022.
For natural lakes, go to the deepest point in the lake to locate the index site (or middle of the lake for
reservoirs). If the deepest point exceeds 50 m in depth, do not establish the index site at this location;
instead, choose a point as close to the middle of the lake as you can without exceeding 50 m in depth.
The procedure below outlines sonar operation and procedures for finding the index site.
For reservoirs, the index site is located near the mid-point of the reservoir rather than at the deepest
point, which may be near the dam. If this would result in an index site that is very shallow or otherwise
non-representative of the reservoir as a whole, choose a point near the center of a main basin, where
depths and the water column will be more representative of the reservoir.
Once in the general area, use the sonar unit to locate the deepest point (< 50 m). When an acceptable
site is located, anchor the boat. Lower the anchor slowly to minimize disturbance to the water column
and sediment. Determine the coordinates of the index site by GPS (if satellite coverage is available) and
record on the Index Samples form. In addition, select if 3 or fewer or 4 or more satellites were used. If
satellite coverage is not available at that time, try again before leaving the index site. The following is
the procedure to be used to locate the index site:
1. Operate sonar unit according to manufacturer's specific operating procedures. If possible, depth
readings should be made and recorded in metric units (be sure to specify units on the Index
Samples form).
2. Use the sonar in the area expected to be the deepest. Spend no more than 30 minutes searching
l/l
for the deepest point; the maximum depth for the index site is 50 meters. til
3. Anchor the boat. >
I
4. Determine the coordinates using GPS. Record GPS coordinates on the Index Samples form. ^
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29
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4.3 Post Sampling Activities
Upon return to the launch site after sampling, review all labels and completed data forms for accuracy
and completeness, and make a final inspection of samples. If information is missing from the forms or
labels, the Field Crew Leader gathers and records the missing information. The Field Crew Leader selects
the red circle at the top of each data form after review. Other post sampling activities include: sample
filtering, inspection and cleaning of sampling equipment, inventory and sample preparation, sample
shipment, and communications.
4.3.1 Equipment Cleanup and Check for Invasive Species
You must inspect all equipment, including nets, boat, and trailer, and clean off any plant and animal
material. This effort ensures that introductions of invasive species such as Eurasian watermilfoil
(Myriophyllum spicatum) quagga mussels (Dreissena rostriformis bugensis), and zebra mussels
(Dreissena polymorpha) do not occur between lakes. Prior to leaving a lake, drain all bilge water or live
wells in the boat. Inspect, clean, and handpick plant and animal remains from vehicle, boat, motor, and
trailer that contact lake water. Inspect and remove any remnants of vegetation or animal life. Before
moving to the next lake, if a commercial car wash facility is available, thoroughly clean vehicle, boat, and
trailer (hot water pressurized rinse - no soap). Rinse equipment and boat with 1% bleach solution or
other approved biological disinfectant to prevent spread of exotics. Procedures are below.
1. Clean for biological contaminants (e.g., Eurasian watermilfoil, zebra mussels, and alewife):
a. Prior to departing from a lake, drain all bilge water from the boat.
b. At the lake, inspect motors, boat, and the trailer for evidence of plant fragments,
especially in or near the propeller and water intakes. Remove all plant fragments.
c. At the lake or base site, dry out and inspect nets and buckets and remove any remnant
vegetation or animal life. Disinfect gear with 1% bleach solution or other approved
biological disinfectant.
d. If a commercial car wash facility is available, thoroughly clean vehicle and boat (hot
water pressurized rinse-no soap).
2. Clean and dry other equipment prior to storage:
a. Rinse chlorophyll-o collection bottle three times with Dl water after each use.
b. Rinse graduated cylinders, integrated sampler, and other sampling devices three times
with Dl water after each use.
c. Briefly soak zooplankton nets and fishing nets in a 1% bleach solution or other approved
biological disinfectant and dry after each use. Do not dry in sunlight because the mesh is
photosensitive.
d. Rinse coolers with water to clean off any dirt or debris on the outside and inside.
e. Rinse boots and waders with water to clean off any dirt, debris, or biological
contaminants on the outside and inside.
3. Inventory equipment and supply needs and request supplies via the electronic Request form
(forms or site kits) or from the FLC (other items or urgent requests).
4. Remove multi-parameter probe meter and GPS from carrying cases and set up for pre-departure
checks and calibration. If present (i.e., not using optical DO sonde), examine the oxygen
membranes for cracks, wrinkles, or bubbles. Replace if necessary.
5. Recharge/replace batteries as necessary.
6. Recheck field forms from the day's sampling activities. Make corrections and completions where
possible, and initial each form after review.
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4.3.2 Shipment of Samples and Forms
You must ship or deliver time-sensitive samples (i.e., all chilled samples) to the appropriate analytical
laboratories as soon as possible after collection and no later than the day after collection. These samples
will be shipped in two groups for next day delivery. Other frozen and non-chilled samples may be
shipped or delivered in batches provided they can be adequately stored. Report all sample shipments to
the NARS IM Coordinator (by submitting the Tracking form in the NLA App) as soon as possible so that
and they can be tracked if they do not arrive when expected.
Field crews are to fill out the pertinent section(s) of the Tracking for each sample shipment. In each
section of the tracking form, the following information must be recorded:
Package tracking number
Date sample(s) were sent
Sample type code:
> ENTE - Fecal Indicator (Enterococci)
> BENT - Benthic macroinvertebrates
> CHEM-Water Chemistry
> CHLX - Chlorophyll-o
> FDNA-eDNA (index)
> LDNA-eDNA (littoral)
> MICZ- Algal toxin (microcystins and cylindrospermopsin)
> PHYX - Phytoplankton
> TRIA - Atrazine Pesticide Screen
> ZOCN - Zooplankton coarse (150-micron mesh)
> ZOFN - Zooplankton fine (50-micron mesh)
> FTIS - Human health fish tissue sample
Sample ID number from preprinted sample label
Number of containers for each sample
Any additional shipping comments
See APPENDIX B: SHIPPING GUIDELINES for further information.
4.3.3 Communications
After the Field Crew Leader has reviewed form content at the end of the sampling day, click the SUBMIT
menu button and choose the form(s) that you wish to submit. After you have chosen which form to
submit, click the green submit button at the bottom of the form list. An email will pop up on your device
addressed to NARSFieldData@epa.gov. Copy yourself and any other crew members or managers and
click send. To ensure that the email was sent, check the SENT mailbox on your email App and look for
the recent email containing the data. If the email is not in the SENT mailbox, it was not sent and you
should try again after verifying an internet connection.
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After each submission, a data summary email will be sent to the email address which submitted the
data. This data summary contains a list of the data forms and their most recent submission date/time as
well as a list of the most critical data points collected at the site. The Field Crew Leader should review
this data summary to ensure that the data forms were successfully received and that critical values are
present and correct.
At any point, if it is determined that data needs to be revised or updated, crews should feel free to do so
in the App and re-submit any edited data forms using the steps above. Newly revised data will
supersede previous data. It is not necessary to re-submit data forms that were unchanged.
Each field crew must submit the Verification form in the NLA App after each site visit (whether the site
is sampled or not). General communications information, including contact information for the NARS IM
Center, is outlined in Section 2.1.
If the field crew documented the potential occurrence of a cyanoHAB event at one or more station in
the lake, the Field Crew leader is responsible for reporting the bloom to the appropriate state, tribal
and/or watershed organization. See Section 2.2.4.5 for potential reporting mechanisms.
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5.0 INDEX SITE ACTIVITIES
Field crews will collect several different measurements and indicators at the index site (as described in
Table 1-1): cyanoHABs assessment, a temperature, DO, and pH depth profile, secchi transparency, fish
eDNA, chlorophyll-o, phytoplankton, algal toxins, water chemistry, atrazine, and zooplankton samples. A
detailed description of the individual elements is provided below.
5.1 CyanoHABs Visual Assessment
When arriving at the index site, field crew views and documents visual observations of a potential
cyanobacterial bloom in the NLA App and photo documents the bloom according to the reporting
mechanism selected by the field crew (i.e., bloomWatch or state-specific reporting mechanism).
5.2 Temperature, DO, and pH profile
5.2.1 Summary of Method
Use a multi-parameter water quality meter (or sonde) to measure temperature, DO, and pH at
predefined depth intervals. Measurement intervals for the profile are based on the site depth (see
Section 5.2.3). Once the profile is completed, make another DO measurement at the surface and
compare it to the initial reading to see if the probe is functioning correctly and holding calibration. If the
lake is thermally stratified, note the top and bottom of the metalimnion based on the temperature
readings (observed as a change of >1 ฐC per meter of depth).
The meters and probes are delicate; take care to avoid putting the probe into contact with sediments
which could foul the probes. An accurate measure of the site depth will help prevent contact between
the sediment and the probes.
5.2.2 Equipment and Supplies
Table 5-1 is the checklist for equipment and supplies required to conduct protocols described in this
section.
Table 5-1 Equipment and supplies - temperature, pH, and DO profiles.
Type
Item
Quantity
Forms
NLA Profile Calibration
1
NLA Index Profile
1
Collection:
Depth Finder (hand-held or boat mounted sonar)
1
Water column
Sounding line (50 m, calibrated, marked in 0.5 m intervals) with clip OR
1
depth
Sounding rod (calibrated) for very shallow lakes
Collection:
Multi-parameter water quality meter (with temperature, pH, and DO probes)
1
Profile
Sounding line or 50-meter tape
measurements &
Sounding weight
1
calibration
Squirt bottle (1 L Nalgene) - De-ionized (Dl)
1
Squirt bottle (1 L Nalgene) - lake water
1
Calibration cups and standards
1
QC check solution
Barometer or elevation chart to use for calibration
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5.2.2.1 Multi-parameter Sonde
The multi-parameter sonde must be heavy enough to minimize sway and wobbling as it is lowered and
raised in the water column. The instrument must be stabilized prior to taking a reading. Experiment with
the sonde prior to sampling and add weight to the cable if needed.
5.2.2.2 Temperature Meter Calibration
Check the accuracy of the sensor against a thermometer (a non-mercury type is recommended) that is
traceable to the National Institute of Standards and Technology (NIST) at least once per sampling
season. The entire temperature range encountered in the NLA 2022 should be incorporated in the
testing procedure and a record of test results kept on file.
5.2.2.3 DO Probe Calibration
Crews are to calibrate the DO probe at the lake prior to each sampling event. The probe calibration is to
be completed in the field against an atmospheric standard (ambient air saturated with water, or water
saturated with air for optical probes) prior to launching the boat. In addition, manufacturers typically
recommend periodic comparisons with a DO chemical analysis procedure (e.g., Winkler titration) to
check accuracy and linearity. Small "mini-Winkler" titration kits are suitable for this check and can be
taken into the field.
5.2.2.4 pH Meter Calibration
Calibrate the pH electrode daily, prior to each sampling event in accordance with the manufacturer's
instructions and your organization's existing SOP. Conduct a QC check with a different standard to verify
the calibration and periodically evaluate instrument precision (see Section 2). Ideally, a quality control
solution (QCS) should be used that is similar in ionic strength to the lake water samples you will be
measuring. Standard buffer solutions used to calibrate electrodes may not be representative of typical
lake waters.
5.2.2.5 Conductivity Calibration
A field conductivity measurement is optional for the NLA 2022. If the Field Crew opts to take
conductivity measurements, the conductivity meter must be calibrated prior to each sampling event.
Calibrate the meter in accordance with the manufacturer's instructions. Ensure that the conductivity
meter is temperature corrected to 25 ฐC.
5.2.3 Depth Profile Procedure
Below are the step-by-step procedures for measuring temperature, pH, and DO profiles at the index site.
1. Calibrate Instrument
a. Check meter and probes and calibrate according to manufacturer's specifications.
b. Enter calibration information on the Profile Calibration form.
2. Record Site Conditions:
a. Observe site conditions and fill out the "Site Conditions" portion of the Index Samples
form. Conditions to be reported include:
i. Precipitation ("None", "Light," or "Heavy")
ii. Surface conditions ("Flat," "Ripples," "Choppy," or "Whitecaps")
b. Presence or absence of odor or scum. If present, select "Yes" and describe the odor or
scum.
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3. Determine Site Depth:
a. Accurately measure the depth using a sounding line with a weight or other means and
record on the Index Samples form.
b. Indicate method used.
4. Determine Measurement Intervals:
a. The number of readings and the depth intervals taken depends on the site depth. Below
is a list of rules for determining the intervals:
i. The profile will always begin with a measurement just below the surface (e.g.,
approximately 10 cm or the minimum depth required to keep all probes
submerged).
ii. The last (deepest) measurements will always be at 0.5 m above the bottom.
iii. If the site is < 3.0 m deep, record measurements beginning just below the
surface and at 0.5 m intervals, until 0.5 m above the bottom.
iv. If the depth is between 3.0-20 m (inclusive), record measurements beginning
just below the surface and then at 1.0 m intervals until reaching 0.5 m above
the bottom.
v. If the depth exceeds 20 m, record measurements beginning just below the
surface, then at 1.0 m intervals until you reach 20 m, then at 2.0 m intervals
until 0.5 m above the bottom.
vi. If the metalimnion is encountered (observed as a change of >1 ฐC per meter of
depth), take measurements at least every meter within the metalimnion.
5. Measure Temperature, DO, and pH:
a. Lower the sonde in the water and measure the vertical profile of temperature, DO and
pH at the predetermined depth intervals. Be careful not to let the probe touch the
bottom.
b. Record the intervals and measurements on the Profile Data form.
c. Use the provided comment bubble to provide extra information about any
measurements that the crew feels needs further comment or when a measurement
cannot be made.
6. Duplicate Surface DO Measurement
a. When the profile is completed, take another DO measurement at the surface, record it,
and compare it to the initial surface DO reading.
b. Mark 'Yes' or 'No' on the form if the second DO reading is within 0.5 mg/L of the initial
surface reading. This provides information regarding measurement precision and
possible calibration drift during the profile.
i. If measurement is not within 0.5 mg/L, verify your calibration.
ii. If DO is found to be out of calibration, re-calibrate and re-record DO
measurements.
7. Determine the Metalimnion:
a. If the lake is thermally stratified, note the top and bottom of the metalimnion in the
Metalimnion column.
b. For standardization purposes, the metalimnion has been defined in the protocol as an
area where water temperature changes at least 1 degree Celsius per meter.
c. If you suspect that the metalimnion exists but does not change at the specified rate,
estimate the top and bottom of the metalimnion to the best of your ability, use the
provided comment bubble to flag the data, and explain.
d. In deep sites where measurement intervals are 2.0 meters apart, decrease the interval
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to 1.0 meters while taking measurements within the metalimnion.
5.3 Secchi Disk Transparency
5.3.1 Summary of Method
A Secchi disk is a black and white patterned disk used to measure a lake's clarity (see Figure 5-1). Take
the reading on the shady side of the boat, without sunglasses, hat, or other viewing aids. Record the
depths where the disk disappears when descending and reappears when retrieving.
Figure 5-1 Secchi disk diagram (USEPA, 1991).
5.3.2 Equipment and Supplies
Table 5-2 is the checklist for equipment and supplies required to conduct protocols described in this
section.
Table 5-2 Equipment and supplies - Secchi disk transparency.
Type
Item
Quantity
Form
NLA Index Samples
1
Collection
Metric tape measure
1
Secchi Disk (20 cm diameter)
1
Sounding line (50 m, calibrated, marked in 0.5 m intervals) with clip
1
5.3.3 Procedure for Determining Secchi Transparency
Because different people measuring Secchi transparency at the same site may obtain different results
(due to differences in vision and interpreting disk disappearance and reappearance), it is recommended
that one crew member conduct Secchi disk measurements at all lakes.
If the lake is shallow and the water clear, the Secchi disk might reach the bottom and still be visible. If
this is the case, it is important to not stir up the bottom sediments while anchoring the boat. Move the
boat away from the anchor before taking the reading. If the disk is visible at the bottom of the lake,
indicate this by filling in the "clear to bottom" check box on the Index Samples form and record the
water depth in both the disappearance and reappearance fields.
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States that wish to take additional measurements for comparisons using a view scope are encouraged to
do so after completing the Secchi disk measurements are completed following the NLA protocols.
The following procedure is to be followed when collecting NLA Secchi disk measurements:
1. Confirm that the lowering line is firmly attached to the Secchi disk.
2. Remove sunglasses and hat. Also, do not use view scopes or other visual aids. If wearing
prescription sunglasses, temporarily replace them with regular clear lens prescription glasses.
3. Lower the Secchi disk over the shaded side of the boat until it disappears.
4. Read the depth indicated on the lowering line. If the disappearance depth is <1.0 meter,
determine the depth to the nearest 0.05 meter by marking the line at the nearest depth marker
and measuring the remaining length with a tape measure or meter stick. Otherwise, record the
disappearance depth to the nearest 0.1 meter. Record the disappearance depth on the Index
Samples form.
5. Lower the disk a bit farther and then slowly raise the disk until it reappears and record the
reappearance depth, using the same level of precision as before.
6. Calculate the euphotic zone depth by multiplying the depth where the Secchi disk reappears by
two. Use this calculation to determine the depth at which water samples will be taken with the
integrated water sampler:
6.1 If euphotic zone is less than 2 meters, water samples will be collected only
within the euphotic zone.
6.2 If euphotic zone is greater than 2 meters, water samples will be taken from the
top 2 meters of the water column.
6.3 If the Secchi is clear to the bottom and the lake is less than 2.5 m deep, water
samples will be collected 0.5 m from the depth at the index site.
7. Record the depth that will be targeted for the integrated water samples.
8. Note any conditions that might affect the accuracy of the measurement in the comments field.
5.4 eDNA Sample Collection - Index
5.4.1 Summary of Method
Collect water samples for eDNA with a grab sample collected at the water surface from the boat.
5.4.2 Equipment and Supplies
Table 5-3 provides the equipment and supplies needed for field operations to collect fish eDNA samples
at the index site.
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Table 5-3 Equipment and supplies - index eDNA samples.
Type
Item
Quantity
Form
NLA Index Samples
1
Documentation
Label: FDNA - Index
Clear tape strips (to cover sample labels)
1
As needed
Collection
Index eDNA bottle (1 L clear, square)
Gloves (latex/nitrile, non-powdered)
1
1 pair
Storing and preserving
Wet ice
Cooler
Plastic electrical tape
As needed
1
As needed
5.4.3 Sampling Procedure
While anchored at the index site, collect a grab sample from the water surface using the 1 liter index
eDNA bottle.
1. Make sure all necessary data has been written on the sample label, the label is placed
on the outside of the bottle, and the label is completely covered with clear tape.
2. Put on surgical gloves (non-powdered). Do not handle any food, drink, sunscreen, or
insect repellant until after the water samples have been collected, or implement
measures to reduce contamination by such chemicals, if applied, such as washing,
wearing long gloves, etc.
3. Remove the cap from the bottle.
4. Do not rinse the bottle and avoid touching the inside of the bottle or the inside of the
cap.
5. Dip the sample container into the surface water (do not fully submerge mouth of
bottle). At the surface of the water, angle the mouth of the sample container away from
the boat. Collect any surface scums, if present, at the site.
6. Fill the 1 L bottle to the 1000 mL mark, leaving headspace for air. This headspace is
important since the sample will be frozen.
7. If the bottle is filled above the 1000 mL mark, discard excess water.
8. Carefully replace the cap. Seal the cap with plastic electrical tape.
9. Immediately after sample is collected, place in a cooler with ice to minimize exposure to
light and begin chilling the sample.
10. Upon return to the vehicle or a base location, freeze the sample. The sample will be sent
frozen with dry ice.
5.5 Water Sample Collection and Preservation
5.5.1 Summary of Method
Collect water samples using an "integrated sampler", which is based on a design by the Minnesota
Pollution Control Agency (MPCA) (see
Figure 5-2). The device is a PVC tube 6.6 feet (2 meters) long with an inside diameter of 1.24 inches (3.2
centimeters) fitted with a stopper plug on one end and a valve on the other. The device allows collection
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of water from the upper two meters of the water column (e.g., within the euphotic zone). If the
euphotic zone is < 2.0 m deep (as calculated in the Secchi Disk Transparency section of the form), lower
the integrated sampler only to the depth of the euphotic zone, and take additional grab samples as
necessary to collect the total volume needed for the samples.
5.5.2 Equipment and Supplies
Table 5-4 provides the equipment and supplies needed to collect water samples at the index site.
Table 5-4 Equipment and supplies - water samples.
Type Item Quantity
Form
NLA Index Samples
1
Documentation
Collection
Labels: water chemistry, algal toxins, phytoplankton,
atrazine, and chlorophyll-o
Clear tape strips (to cover sample labels)
Integrated sampler device (MPCA design)
Funnel
Gloves (latex/nitrile, non-powdered)
1 per
sample
As needed
1
1
1
Storing & Preservation
Cubitainerฎ (4L) - water chemistry
HDPE bottle (60 mL, white, wide-mouth) - atrazine
HDPE bottle (500 mL, white, round) - algal toxins (MICZ)
HDPE bottle (1 L, white, narrow-mouth) - phytoplankton
Poly bottle (2 L, brown) - chlorophyll-o
Wet ice
Lugol's solution (250 mL bottle)
Cooler
Plastic electrical tape
1
1
1
1
1
As needed
5-10 mL
1
As needed
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Stopper
Float Material
ฆ PVC Pipe
I.D. 1.25 inches
(3.2 cm)
- Ball Valve
Figure 5-2 Integrated water sampler device (MPCA).
5.5.3 Sampling Procedure
Assuming the euphotic zone is > 2 meters and the depth of the lakes is greater than 2.5 m, collect four
integrated water samples (Figure 5-3). Sample #1 is emptied into a 2 L sample bottle for chlorophyll-a
filtering. Sample #2 is to be transferred from the sampler to the 4 L Cubitainerฎ, mixed thoroughly, and,
poured off into one 1 L sample bottle for phytoplankton processing, one 500 mL bottle for the algal
toxins samples, and one 60 mL bottle for the atrazine pesticide sample. Samples #3 and #4 are to be
transferred from the sampler to the 4 L Cubitainerฎ for the water chemistry sample. If the euphotic
zone is <2 meters, only collect water from the euphotic zone and increase the number of grab samples
accordingly. Additionally, if the secchi is clear to the bottom and the lake is less than 2.5 m deep, water
samples will be collected 0.5 m from the depth at the index site.
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Integrated
Sampler #1
a
Chlorophyll-a
Integrated
Sampler #2
Atrazine
1 L
Phytoplankton
4 L
500
mL
Algal toxins
Discard remaining water from
sampler#2 before collecting
samplers #3 and #4
Integrated
Samplers #3 and #4
4 L
Water Chemistry
Figure 5-3 Procedure for using the integrated sampler device to collect depth integrated samples.
5.5.3.1 Sample Collection
1. Make sure all necessary data has been written on the sample labels and labels are
completely covered with clear tape.
2. Put on surgical gloves (non-powdered). Do not handle any food, drink, sunscreen, or insect
repellant until after samples have been collected, or implement measures to reduce
contamination by such chemicals, if applied, such as washing, wearing long gloves, etc.
3. Rinse each water sample collection container and lid with surface water three times.
4. Remove the rubber stopper cap and open the valve on the bottom end of the integrated
sampler. Rinse by submerging it three times in the lake and draining after each rinse.
Complete rinsing on the opposite side of the boat from which you plan to sample. Do not ^
LU
take samples near the motor or other sources of contamination. j=
5. Slowly lower the sampler into the lake as vertically as possible. For a 2 m sample, stop ^
lowering the device when the upper end is just above the surface. If the euphotic zone is < <
LU
2.0 m deep (as calculated in the Secchi Disk Transparency section of the form), the t
integrated sampler will be lowered only to the depth of the euphotic zone; additional ><
41
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samples will be taken to collect the volume needed for the samples (8 L total). In clear
shallow lakes, samples will be collected 0.5 m from the depth at the index site.
6. Cap the upper end with the rubber stopper firmly and slowly raise the sampler.
7. When the bottom of the sampler is near the surface, reach underwater and close the valve
on the bottom end.
8. Lift the sampler into the boat, keeping it as vertical as possible. When possible, move the
containers to a shaded area of the boat to avoid exposing the sample to direct sunlight
when dispensed.
9. Carefully open the valve and dispense the contents of sample #1 (or more, as necessary to
fill bottle**) into the 2 L brown bottle. This is the chlorophyll-o sample, which will be filtered
on shore (see Section 8.2). Place the bottle on ice until filtration can be initiated.
**ifthe last sample pull does not completely fit in the container, homogenize the sample
in the integrated tube prior to dispensing
10. Repeat the collection process in steps 5-8 and dispense the contents of sample #2 (or more,
as necessary) into the 4 L Cubitainerฎ and mix well.
11. Fill the 1 L phytoplankton bottle from the 4 L Cubitainerฎ, allowing enough headspace to
add at least 5 mL of Lugol's preservative.
12. Fill the 500 mL bottle from the 4 L Cubitainerฎ, leaving headspace so that the bottle doesn't
burst when frozen. This is the algal toxin sample. Seal the cap with plastic electrical tape.
Place the bottle in the cooler with ice.
13. Fill the 60 mL bottle from the 4 L Cubitainerฎ to just below the shoulder, leaving headspace
so that the bottle does not burst when frozen. This is the atrazine sample. Seal the cap with
plastic electrical tape. Place the bottle in the cooler with ice.
14. Discard the residual water in the Cubitainerฎ.
15. Pour the contents of sample #3 and sample #4 (or more, as necessary) from the integrated
sampler into the 4 L Cubitainerฎ, removing as much air from the Cubitainerฎ as possible.
This is the water chemistry sample. Seal the cap with plastic electrical tape. Place the
Cubitainerฎ in the cooler with ice.
1. All samples are to be kept in a cooler on wet ice until preservation as needed.
2. For the phytoplankton sample, add 5 mL of Lugol's solution to the 1 L phytoplankton bottle.
(Fill the provided plastic transfer pipette up to the bulb with Lugol's three times to get 5
mL). Cap the bottle and invert until well mixed. The sample should resemble the color of
weak tea. If needed, add additional Lugol's 2-3 mL at a time up to a maximum of 10 mL. Seal
the cap with plastic electrical tape.
3. Upon return to a base location with a freezer or portable freezer, freeze the algal toxin and
eDNA samples. These samples will be sent frozen with dry ice.
5.5.3.2 Sample Preservation
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5.6 Zooplankton Collection
5.6.1 Summary of Method
Collect two vertical samples using a fine mesh (50 pirn) and coarse mesh (150 pirn) Wisconsin nets with
collection bucket attached at the cod end. Each net is slowly lowered over the side of the boat into the
water. The net is retrieved back to the surface at a slow, steady rate. Lift the net out of the water; rinse
it from the outside to free organisms from the side of the net and to concentrate them in the collection
bucket. Narcotize the organisms with an effervescent sodium bicarbonate tablet (e.g., Alka-Seltzerฎ
tablet). Transfer the sample from the collection bucket to a 125 mL sample container and preserve each
sample with 95% ethanol. You will repeat the procedure with the other net on the opposite side (or end)
of the boat. The cumulative tow length for each net is 5 m. In shallow lakes, multiple tows with each net
are required to achieve the cumulative tow length. The objective is to sample a sufficient volume of
water to obtain at least 300 organisms per sample from all but the most oligotrophic lakes.
5.6.2 Equipment and Supplies
Table 5-5 provides the equipment and supplies needed to collect a zooplankton sample. Figure 5-4 is an
illustration of the zooplankton nets and collection buckets.
Table 5-5 Equipment and supplies - zooplankton collection.
Type
Item
Quantity
Form
NLA Index Samples
1
Documentation
Labels: zooplankton course and zooplankton fine
Clear tape strips (to cover sample labels)
1 per sample
As needed
Collection
Plankton net fine (50 nm) and collection bucket
Plankton net course (150 nm) and collection bucket
Sounding line (50 m, calibrated, marked in 0.5 m
intervals) with clip
Funnel
Squirt bottle (1 L Nalgene) - de-ionized (Dl)
Squirt bottle (1 L Nalgene) - lake water
Effervescent sodium bicarbonate (Alka seltzer)
tablets
Pail (narcotization and concentration chambers)
1
1
1
1
1
1
1
1
2
Storing & Preservation
HDPE bottle (125 mL, white, wide-mouth)
Ethanol (95%)
Plastic electrical tape
2
1
As needed
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ATTACHMENT
REDUCING
COLLAR
METAL RETAINING
BANDS
(Not to be removed)
THREADED
ซ REMOVABLE
BUCKETS
Figure 5-4 Wisconsin net and collection bucket diagram,
5.6.3 Sampling Procedure
The procedures for collecting and processing zooplankton samples are presented below.
5.6.3.1 Sample Collection
1. Determine and record the number of tows required to achieve the standard cumulative 5 m tow
on the Index Samples form.
a. For lakes deeper than 7 m, you will take a single 5 m tow with each net.
ฃ b. For lakes with a depth less than 7 m, you will determine and record the number of tows
b that will be required to achieve a standard cumulative 5 m tow for each net (Table 5-6).
g For example, if the lake is 6 meters deep, you will take two 2.5 m tows with each net. All
< lakes less than 7 m deep require at least two tows because the collection bucket should
I i I
H never touch the sediment due to potential fouling.
x
LU
O
44
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Table 5-6 Lengths and numbers of zooplankton tows based on Index Site depth
Depth of lake (m)
Length of Tow
Number of Tows
7 or more
5 m
1
4 to <7
2.5 m
2
2 to <4
1 m
5
less than 2
0.5 m
10
c. The zooplankton collection methods vary slightly depending on the number of tows
required to achieve a standard cumulative 5 m tow.
i. If the number of tows = 1: follow steps 2 through 13 described below.
ii. If the number of tows > 2, follow steps 2 through 12 described below. After step
12, pour the contents of the collection bucket into a clean (i.e., Dl rinsed) 1-
gallon pail. Rinse the collection bucket with Dl. While taking care not to tip the
zooplankton sample in the pail, repeat steps 2 through 12 for the second tow.
Add the contents of the collection bucket from the second tow to the pail.
Continue to take zooplankton tows and add samples from the collection bucket
into the pail until the target number of tows (2, 5, or 10) is reached. On the last
tow, pour the contents of the pail into the collection bucket to filter the excess
water. Rinse the bucket with Dl water and pour the contents of this rinse into
the collection bucket with the zooplankton sample. Once the zooplankton
sample has been filtered down to an appropriate volume in the collection
bucket, continue on to step 13.
2. Prior to each use, carefully clean and thoroughly rinse the interior of the plankton nets and
buckets with Dl water.
3. Carefully inspect the nets and buckets for holes or tears.
4. Attach the collection buckets to the "cod" end of the nets and secure. Make sure the correct
bucket is attached to the correct net (i.e., the mesh sizes match).
5. Attach the bridled end of the plankton net to a 0.25" calibrated line with markings every 0.5 m
(use the same line as was used with the Secchi disk).
6. Carefully and slowly lower the first net in a constant upright position over the side of the boat.
7. Continue lowering the net to the correct depth (remember to account for the length of the
bridle). If more than one tow is needed, be sure to take additional tows from different locations
around the boat.
8. Retrieve the net by pulling back to the surface at a steady rate (0.3 m or 1 ft/s) without
stopping.
9. Once at the surface, slowly dip the net up and down in the water without submersing the net
mouth to rinse contents into the collection bucket.
10. Complete the rinsing of the net contents by spraying lake water against the outside of the net
with a squirt bottle or similar tool. Be careful not to splash or squirt lake water into the net
mouth, or additional organisms may be collected.
11. If additional rinsing is needed on the interior of the net, use a squirt bottle with Dl water only to
avoid introducing additional organisms.
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12. Once all organisms have been rinsed into the collection bucket, hold the collection bucket in a
vertical position, and carefully remove the bucket from the net.
13. Repeat steps 5-12 with the second net on the opposite side (or end) of the boat.
1. Set the collection buckets in a pail filled half full of lake water to which 2 C02 (e.g., Alka-Seltzer)
tablets have been added outside of the collection buckets. Wait 30 to 60 seconds for the C02
tablets to dissolve before placing the collection buckets into the water. Ensure that the
organisms in the collection buckets are submerged in the water, but be careful not to submerge
the top of the collection buckets, or sample loss will occur. The C02 narcotizes the zooplankton
to relax their external structure prior to preservation in 95% ethanol. This facilitates taxonomic
identification. Wait until zooplankton movement has stopped (usually about one minute).
Raising and lowering the collection buckets in the pail can help water exchange within the
bucket.
2. Check the sample label on the bottle to verify which sample has been collected (coarse or fine
mesh).
3. Use small volumes of Dl water from a squirt bottle to rinse the contents of the mesh net
collection bucket into the 125 mL polyethylene bottle. Rinse the collection bucket with Dl water
three to four times or until the majority of zooplankton have been removed without allowing
the bottle to fill more than half full (~60-70 mL of sample and rinse water combined). After the
zooplankton have been transferred and the sample bottle is half full with sample and rinsate, fill
the bottle to the shoulder with 95% ethanol. Use a funnel, if necessary, when transferring the
sample, rinsate or ethanol to the 125 mL sample bottle
4. In some cases, the volume of zooplankton collected in the collection bucket may exceed 125 mL.
Under this scenario do not try to force the entire sample into a single bottle, or the preservative
will not function properly and the sample may be lost. In such cases, fill the first bottle half full,
and then use a second bottle to preserve the additional amount of sample. Use an "extra jar"
label (i.e., one with no sample ID pre-printed on it) for identification purposes. Complete the
label and print in the sample number assigned to the first container on the label of the second
container. On the form, record a "2" in the "No. Jars" field.
5. On the Index Samples form, fill in the bubble to indicate that the sample is preserved.
6. Verify that all information on the labels and the form is complete and correctly recorded.
7. Repeat steps 2-6 for the second sample collected.
5.6.3.2 Sample Processing
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6.0 LITTORAL AND SHORELINE ACTIVITIES
To better understand near-shore habitats and their condition, travel to 10 evenly spaced physical
habitat (PHab) stations around the lake and document conditions and characteristics observed within
defined plot areas. The full set of measurements, observations, and sampling described in this chapter
water sample for eDNA analyses at each of the 10 stations;
measures or observations of physical habitat cover and structure in the littoral, shoreline, draw-
down, and riparian zone plots at the 10 PHab stations;
visual assessment of a cyanobacterial bloom;
sampling of benthic macroinvertebrates at each of the 10 stations; and
water sample for Enterococci at the last station sampled (for lakes 10,000 ha or less) or boat
launch (for lakes >10,000 ha).
For lakes with a surface area of greater than 10,000 ha (defined as large lakes), crews are not required
to travel to the 10 PHab stations to make physical habitat measurements and collect benthic
macroinvertebrate samples. The requirement was waived on large lakes because of the increased level
of effort required to travel around the shorelines of these large lakes. Nevertheless, we encourage crews
to complete the physical habitat characterizations and macroinvertebrate collections on large lakes, just
as they are done on smaller lakes, so that large lake physical habitat information can be included in the
national assessment.
Note: When large islands are present, more than 10 PHab sites will be identified and assessed for a lake.
6.1 Littoral eDNA Sample Collection
6.1.1 Summary of Method
Collect a grab sample for eDNA at each of the 10 shoreline PHab stations. Grab samples are collected at
the water surface from the boat and added to a single 1L littoral composite sample. If more than 10
PHab stations are identified for the lake, littoral eDNA samples will only be collected from the 10
standard PHab stations around the perimeter of the lake. For "large lakes" (greater than 10,000 ha) the
entire 1L sample is to be collected from the launch site at the end of the day with the enterococci
sample.
includes:
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6.1.2 Equipment and Supplies
Table 6-1 provides the equipment and supplies needed for field operations to collect the littoral eDNA
sample.
Table 6-1. Equipment and supplies - fish littoral eDNA samples.
Type
Item
Quantity
Form
NLA Littoral Samples
1
Documentation
Label: LDNA- littoral
1
Clear tape strips (to cover sample labels)
As
needed
Collection
Littoral eDNA bottle (1 L clear, square)
1
Littoral eDNA collection bottle (125 mL clear, square)
1
Gloves (latex/nitrile, non-powdered)
1-2 pairs
Storing and preserving
Wet ice
Cooler
As
needed
Plastic electrical tape
1
As
needed
6.1.3 Sampling Procedure
When arriving at the PHab station, prior to the collection of PHab and benthic macroinvertebrate
sample , collect a 100 mL littoral eDNA sub-sample from an undisturbed area in the littoral plot. Sub-
samples from all stations are added to the 1 L littoral eDNA composite bottle.
1. At the first PHab station, make sure all necessary data has been written on the littoral
eDNA label, the label is placed on the outside of the 1L littoral eDNA bottle, and the label is
completely covered with clear tape. Samples from 10 physical habitat stations will be added to
this bottle.
2. Put on surgical gloves (non-powdered). Do not handle any food, drink, sunscreen, or insect
repellant until after the water samples have been collected, or implement measures to reduce
contamination by such chemicals, if applied, such as washing, wearing long gloves, etc.
3. Identify an area in or near the littoral plot that is about 1 m deep ("waist deep) to collect the
sample. Avoid sampling shallower water and water at the shoreline due to potential
sediment contamination. If the entire littoral plot is less than 1 m deep, you may collect the
sample outside the littoral plot (e.g., as you are approaching the plot from a location that is
about 1 m deep).
4. Remove the cap from the 125 mL collection bottle. Avoid touching the inside of the bottle or the
inside of the cap.
6. Dip the sample container into the surface water (do not fully submerge mouth of bottle) in an
undisturbed area of the lake. At the surface of the water, angle the mouth of the sample
container away from the boat or body and fill the collection bottle to the 100 mL mark. Collect
any surface scums, if present, at the site. If more than 100 mL is collected, pour off
excess.Remove the cap from the 1L composite bottle. Do not rinse this bottle and avoid
touching the inside of the bottle or the inside of the cap.
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7. Add the 100 mL sample to the 1 L composite bottle. Cap both bottles and place both bottles in a
cooler with wet ice while transporting to the next station to minimize exposure to light and
begin chilling the sample.
8. REPEAT steps 2-6 at the next PHab station, up to a total of ten stations (1L sample total).
9. At the last (10th) PHab station, ensure the sample does not exceed the 1000 mL mark, leaving
headspace for air. This headspace is important since the sample will be frozen. If the bottle is
filled above the 1000 mL mark, cap the bottle, gently mix 3 times, and discard excess
water down to the 1000 mL mark.
10. Carefully replace the cap. Seal the cap with plastic electrical tape.
11. Immediately place the 1L bottle in a cooler with ice and keep the sample chilled throughout the
sampling day.
12. Discard the 125 mL collection bottle after collecting all 10 subsamples. A new collection bottle
will be sent in each site kit.
13. Upon return to the vehicle or a base location, freeze the sample. The sample will be sent frozen
with dry ice.
**At large lakes (lakes over 10,000 hectares) when littoral sampling is not conducted, the entire 1L
littoral eDNA sample will be collected at the launch site with the enterococci sample.
6.2 Physical Habitat Characterization
6.2.1 Summary of Method
Prior to the sampling visit, determine the approximate locations of the 10 PHab stations and mark them
on a Site Map (see Chapter 3), if applicable. Figure 3-2 shows example placement and distribution of
PHab stations around a lake. At each of the 10 PHab stations, you will set up a plot as shown in Figure
6-1 based on visually estimated dimensions. The plot measures 15 m wide and includes: a littoral plot
extending 10 m out from the shoreline; a drawdown zone plot extending inland from the shoreline to
the normal high-water level; aim shoreline zone band at the shore just above the present water line;
and a 15 m wide riparian plot that begins at the normal high water mark and extends 15 m landward.
The drawdown zone plot extends a variable distance inland depending on the degree of drawdown and,
if the distance from the present shoreline to the normal high water mark is negligible (i.e.,
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As described below, you will record observations from each of the zones on the Physical Habitat form.
6.2.2 Equipment and Supplies
Table 6-2 lists the equipment and supplies needed to locate the PHab stations and conduct the physical
habitat characterization.
Table 6-2 Equipment and supplies - physical habitat assessment.
Type
Item
Quantity
Forms
NLA Verification
1
NLA Physical Habitat
10-12
Collection
Depth Sounder (hand-held or boat mounted sonar)
1
Sounding rod (3 m, marked in 0.1 m increments, calibrated, PVC)
1
GPS unit (with manual, reference card, extra battery)
1
Rangefinder (for estimating horizontal drawdown)
1 (optional)
Clinometer (for use as a level to measure vertical drawdown)
1 (optional)
Surveyors rod (for measuring vertical drawdown)
1 (optional)
50-meter tape (measurements as needed)
1
Binoculars (for making observations of distant riparian)
1
Map wheel or string (for measuring shoreline distances on site map)
1
Anchor (with 75 m line or sufficient to anchor in 50 m depth)
1
Buoy (for marking observation point)
1
6.2.3 Locating the Physical Habitat Stations and Defining the Shoreline Boundary
6.2.3.1 Base Site Activities
It is important that you set up PHab stations in the office to minimize bias in site selection and to ensure
efficient location of stations once at the lake.
1. Using a lake map, select a random starting point on the lake outline. Any reasonable method
may be used select the starting point (e.g., toss a pencil randomly on the map, letting the sharp
end point to the nearest shoreline location). This random starting point is your "A" station.
2. It is important that the remaining nine stations be located at equal distances around the lake
(see Figure 3-2). These will be your "B" through "J" stations. Field crews can do this manually (by
either using a string to trace the perimeter of the lake, which can then be straightened and
marked in equal intervals, or by using a map wheel) or electronically (with GIS or other digital
mapping tools) to measure the perimeter of the lake and divide by 10.
3. Using a GIS or other digital mapping tool application to locate the coordinates of the 10 stations
that can then be entered as GPS waypoints greatly facilitates correctly locating PHab stations by
boat in the field, especially on large lakes.
4. Mark the physical habitat stations on a site map, if applicable.
Note: In revisit lakes (see Section 9.1 for more information), crews will re-randomize and relocate the
PHab stations. The stations are re-randomized because the revisit data is used to examine variability of
the entire lake assessment.
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6.2.3.2 Littoral and Shoreline Activities
Using the site maps and GPS, proceed by boat around the lake, locate, and stop at each of the 10 PHab
stations. Position the boat at a distance of 10 m from shore, anchor if necessary, and make the
measurements and semi-quantitative observations specified on the Physical Habitat form in the NLA
App. Complete a separate Physical Habitat form for each station (see tabs A-L at the top of the form).
Make every reasonable attempt to record physical habitat observations and measurements for all 10
PHab stations. Where physical habitat observation and measurements are impossible, record comments
as specified in Table 3-1.
6.2.3.3 Shoreline and Station Location Adjustments
Once in the field, you may encounter situations that require you to modify the shoreline and/or station
location(s) from the intended locations marked on the site map. If this occurs, make the corrections and
adjustments on the Physical Habitat form and note the reasons in the comments section of the form.
The general guidelines for locating or modifying the location of the littoral and shoreline stations are
summarized below.
1. Locate station using maps, aerial photos, or GPS units.
2. Define shore as either the current waterline OR the boundary between open water and the edge
of dense vegetation (terrestrial, wetland, or emergent vegetation) or extensive very shallow
water (shoreline defined by limit for navigating your boat).
3. If the shoreline observed in the field differs from the mapped shoreline, mark "Station
Relocated" and enter a comment on the Physical Habitat form stating the apparent reason (e.g.,
drought, lake drawdown, flooding, dredging, limited boat access, etc.).
4. If a PHab station is inaccessible because of shoreline changes, mark "Station Relocated" at the
top of the Physical Habitat form, and position one or more new stations at approximately equal
intervals.
5. If a station is eliminated, select the check box at the top of the form.
6. If the shoreline observed in the field differs radically from the site map and you are sure you are
at the correct lake, you can sketch a map of the lake. Use a string to measure the new outline,
divide it into 10 equal parts, and lay out the 10 station locations.
6.2.3.3.1 Islands
Islands may be an additional source of shoreline habitat on a lake and those will be accounted for by
adding island physical habitat stations. Island stations are in addition to the 10 stations (A-J). The
guidelines for adding island stations follow:
ฆ If the combined shoreline of all islands make up 10-20% of the lake's total shoreline, add one
PHab station (stations will now be labeled A through K)
ฆ If the combined shoreline of all islands make up more than 20% of the lake's total shoreline, add
two PHab stations (stations will now be labeled stations A through L)
ฆ Island stations are designated by marking the "Is it an island?" bubble on the form, by a new
station letter (K or L), and by marking the island location and station on a site map.
Island stations, i.e., K and L, should be selected at random (e.g., toss a pencil randomly on a map, letting
the sharp end point to the nearest island location, if needed, repeat to identify station L).
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6.2.3.3.2 Ambiguous Shorelines
The shoreline is defined as the interface between "lake-like" conditions and riparian or wetland
conditions. In most cases, the shoreline will be easily identified as the current waterline. In some
instances, however, the shoreline might not be obvious. Listed below are some general situations and
rules that should be applied to ambiguous shorelines.
ฆ If there has been a significant drop in lake level due to drought, purposeful drawdown, dam
repair, or other reasons, shallow areas may be exposed that are usually covered with water.
In this case, consider the current waterline as shoreline for the purposes of this survey, not
the normal waterline.
ฆ If there are extensive very shallow areas or shoals, consider the shoreline to be the
boundary between the shallow area and deeper open water, as defined by ease of access by
a small sampling boat.
ฆ If access to the true shoreline is prevented by an area of dense aquatic or terrestrial
vegetation, consider the shoreline to be the boundary between the vegetation and deeper
open water. Again, define the operational shoreline by ease of access by a small sampling
boat. This may result in a riparian zone that can be more of a wetland than an upland
vegetation plot.
ฆ If a river or stream enters a lake, the shoreline begins where no flow is visible.
ฆ If there is flooding, try to find the position of the normal high water mark. The normal high
water mark may be evident by the extent of flooded trees or other terrestrial vegetation.
The goal of the physical habitat survey is to characterize the lakeshore based on observations of
conditions at 10 evenly spaced PHab stations around the lake. Adjustments to station locations might be
needed if the field crew runs into unusual conditions or problems. Below are some rules concerning
modifications to the station location(s).
ฆ If only a small portion of the shoreline differs and it does not affect, or only slightly affects, a
PHab site location, sketch the lake shoreline on the site map and reposition the station (if
needed).
ฆ If the difference causes a contraction of the shoreline and a PHab station location is lost,
sketch the lake shoreline on the site map and make a decision to either: (a) keep the station,
relocate it on the revised shoreline map and adjust some or all other stations in order to
keep stations evenly spaced around the lake (i.e., keep 10 stations), or (b) eliminate the
station altogether (i.e., reduce the number of stations).
ฆ If the Site Map does not in any way match the lake shoreline, draw a new sketch map
approximating the shoreline, and re-establish the 10 PHab stations. A quick way to locate 10
evenly-spaced PHab stations is to: (a) lay a piece of string on the lake perimeter, (b) pick up
the string, measure it, and mark out 10 equal parts, and (c) lay the string back on the
perimeter and use the marks to locate the 10 sites on the map.
ฆ If a PHab station is inaccessible, you must make a decision to either: (a) relocate the station
and adjust some or all other stations so that they are evenly spaced around the lake (i.e.,
keep 10 stations), or (b) eliminate the station altogether (i.e., reduce the number of
6.2.3.3.3 Actual shoreline is different than appears on the map
6.2.3.3.4
PHab Station is inaccessible
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stations). The size of the lake will help drive this decision.
ฆ Draw all adjustments to the shoreline based on field observations directly on the Site Map
and explain the adjustments in the comments section of the Physical Habitat form.
6.2.3.3.5 Identifying Relocated and New Stations on the Form
Use the following notations when recording station location modifications.
ฆ If you relocate a station, note the new location on the Site Map and fill in the bubble
corresponding to the original station letter (e.g., "C") on the Physical Habitat form. In
addition, fill in the "Station Relocated" bubble on the form to indicate that the station has
been moved from its originally intended location.
ฆ If a station is lost and cannot be replaced, cross out the original station location on the Site
Map, fill in the bubble corresponding to the original station letter, and fill in the "Dropped"
bubble on the Physical Habitat form.
6.2.4 Establishing the Physical Habitat Plots at each station
Establish a plot for physical habitat characterization at each PHab station by visually estimating the plot
dimensions. Most littoral, shore, and near-shore observations and measurements can be made from the
boat at the observation point 10 m off-shore (estimated by eye). Limit observations at each station to
the area that is within the defined plot dimensions (additional observations of human activities are
made adjacent to or behind the defined plots). After setting plot dimensions, you may need to move
around within the littoral plot to see or probe the bottom, or even go onto shore to make observations.
6.2.4.1 Physical Habitat Plot Dimensions
You will identify up to four distinct zones within each physical habitat plot (Figure 6-1), where you
estimate the zone dimensions by eye or with the aid of a rangefinder.
6.2.4.1.1 Littoral
This within-lake zone is a fixed size that is 15 m wide along the shoreline and extending 10 m offshore
into the lake.
6.2.4.1.2 Shoreline
The shoreline band is a fixed 15 m wide strip along the shore from the present water line to 1 m inland.
If the horizontal drawdown distance is <1 m, the shoreline band is within the riparian plot; if the
horizontal drawdown is >1 m, the shoreline band is within the drawdown plot. The shoreline boundary is
defined as the approximate interface between "lake-like" conditions and riparian or wetland conditions.
In cases where the lake shoreline is not obvious (e.g., where there is evidence of large seasonal change
in lake level), define the shoreline as the current waterline. In cases where the lake shoreline is not
visible, define the lake shoreline as the approximate boundary between open water and swamp or
marsh conditions into which your boat could not easily move.
6.2.4.1.3 Drawdown
Under all circumstances, vertical height and horizontal distance of drawdown or water level fluctuations
are measured or estimated. When horizontal drawdown is >1 m, establish a drawdown zone plot with a
fixed width (15 m) but with a variable extent inland. The inland extent of the drawdown plot is equal to
the horizontal extent of drawdown and may differ among the 10 PHab stations, depending on the
topography at each station. It is determined by your judgment and measurement of the horizontal
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drawdown distance from the shore to the normal high water mark at each station.
The vertical height can be visually estimated or measured using a clinometer as a level in combination
with a survey rod or metric tape measure. Similarly, the horizontal distance up the bank between the
current lake level and the evidence of the normal high water level can be estimated visually, or
measured with with a laser range finder, survey rod or metric tape measure when distances are greater
than approximately 15m.
The intent of the drawdown assessment is to capture and track changes in the magnitude of lake level
fluctuation or progressive decline or drawdown of lake levels relative to the normal high water mark.
The normal high water mark is best indicated by the terrestrial vegetation and substrate present. In
most regions of the US, summertime lake levels will be below the high water mark, so the NLA
assessment may register a small amount of drawdown for most lakes. Negligible drawdown depends, to
some extent, on the size of the lake. For NLA, an assessment of the vegetation and potential fish cover
in the drawdown plot is not required when horizontal exposure of the littoral bottom is lm horizontal drawdown), the riparian zone plot is a 15m x 15m square
just inland of the drawdown plot (i.e., a fixed size 15 m wide along the shoreline and extending 15m
inland from the normal high-water mark).
6.2.4.2 Physical Habitat Station Layout Procedures
6.2.4.2.1 Normal High Water
Using the present shoreline, place the littoral plots lakeward from the current water's edge or
the operational shoreline as defined in previous sections.
Draw or sight a straight line inland perpendicular to the shoreline. If this line does not intersect
with the normal high water mark, move the littoral plot laterally in either direction until the
perpendicular line intersects the normal high water mark. Choose to move the plot left or right
based on the direction which results in the least distance moved.
Establish aim shoreline band at the current water's edge or the operational shoreline.
Establish the riparian plot inland from the current water's edge or the operational shoreline
identified above. If using an operational shoreline, the riparian plot may include shallow water
and/ or impenetrable wetland.
The left and right edges of the riparian plot, the shoreline band, and the littoral plot should all
align with one another.
6.2.4.2.2 Below Normal High Water (Drawdown)
Using the present shoreline, place the littoral plots from the current water's edge or the
operational shroreline lakeward as defined in previous sections.
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2. Draw or sight a straight line inland perpendicular to the shoreline. If this line does not intersect
with the normal high water mark, move the littoral plot laterally in either direction until the
perpendicular line intersects the normal high water mark. Choose to move the plot left or right
based on the direction which results in the least distance moved.
3. Establish aim shoreline band at the current water's edge or the operational shoreline.
4. Establish the riparian plot inland from the normal high water mark identified above.
5. Establish a drawdown plot between the littoral and riparian plots. If the drawdown plot is
greater than 1 m, cover estimates will be assessed in this plot.
6. The left and right edges of the riparian plot, the shoreline band, the littoral plot, and the
drawdown plot should all align with one another.
1. Establish the littoral plots lakeward from the normal high water mark, which may be evident by
the extent of flooded trees or other terrestrial vegetation, lakeward.
2. Establish the riparian plot inland from the normal high water mark. In flooded situations, the
riparian zone might be dry, partly flooded, or completely flooded with lake water.
3. Establish aim shoreline band at the current water's edge. In flooded situations, the shoreline
band might be within or inland from the riparian plot.
4. The left and right edges of the riparian plot, shoreline band, and the littoral plot should all align
with one another.
6.2.5 General Observations
Begin the physical habitat characterization with general observations.
1. Set up your plots within your physical habitat station.
2. Measure and record the lake depth 10 m from the shore at each PHab station (observation
point)
3. Note on the Physical Habitat form whether there is shoreline flooding (i.e., observed water level
presently above the normal high water mark) by checking the box.
a. If flooding is present, try to find the position of the normal high water mark, which may
be evident by the extent of flooded trees or other terrestrial vegetation. Measure the
depth at this point; record this as the "depth of flooding." Measure or estimate the
distance from this normal high water position landward towards the margin of flooding;
this establishes the location of the lm shoreline band and the horizontal distance of
flooding. The riparian plot is located as a 15 x 15 meter square abutting the normal high
water mark, regardless of whether it is dry, partly flooded, or completely flooded with
lake water.
b. If the current water level is not above normal high water, enter zeros for height and
horizontal distance of flooding.
4. Note on the Physical Habitat form whether the horizontal drawdown distance is greater than
lm by checking the box. Regardless of the amount of drawdown of lake level fluctuation,
measure or estimate it by recording the vertical height and the horizontal distance between
the present lake level and the normal high water line. Note that these measurements or
estimates may be zeros if the water level is at or above the normal high-water mark, but should
6.2.4.2.3 Above Normal High Water (Flooding)
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never be left blank. If the horizontal drawdown distance is 75 degrees);
b. steep (>30 to 75 degrees; need hands to climb up);
c. gradual, (5 to 30 degrees; can walk up); or
d. flat (< 5 degrees).
NOTE: Complete this estimate even if there is no drawdown.
6. Record the presence or absence of water surface scums, algal mats, or oil slicks within the
littoral zone.
6.2.6 Estimate Substrate Characteristics
You will estimate and assign percentage areal cover for substrate types (e.g. bedrock, boulders, cobble,
gravel, sand, silt/clay/muck, woody debris, organic matter, and vegetation) and also for fish habitat
cover, aquatic macrophytes, and terrestrial vegetation. The categories are as follows:
ฆ 0 = absent (0% cover)
ฆ 1 = sparse (<10% cover)
ฆ 2 = moderate (10 - 40% cover)
ฆ 3 = heavy (>40 - 75% cover)
ฆ 4 = very heavy (>75% cover)
When estimating cover combinations in the substrate section of the Physical Habitat form,
combinations consider that the combined cover of the various types of substrate should add up to
approximately 100%. Because you are assigning cover percentage categories, look at various
combinations of the high and low end of each class. Accordingly, more than one class might be given
sparse (1), moderate (2), or heavy (3) ratings. One dominant class with no clear subdominant class might
be ranked very heavy (4) with all the remaining classes either sparse (1) or absent (0). Two dominant
classes with more than 40 percent cover can both be given a 3.
Estimate the areal cover of bottom substrate types and particle size classes observed within the littoral
and the shoreline zones. Cover categories range from absent to very heavy. Record all observations by
filling in the appropriate bubbles on the Physical Habitat form. In most cases these estimates can be
made from the boat or can be made while wading, if preferred. Substrate types not present should be
assigned zero cover.
ฆ If the bottom substrate is not visible, you should probe the bottom beneath the boat with
the sounding rod (you may have to move closer to shore if you are too deep to use the rod).
Soft sediment can be brought to the surface for examination. Hard sediments can be "felt"
with the sounding rod. Sandy substrate can be "felt" or "heard" by twisting the sounding rod
and detecting grittiness. Estimating cover of various substrate types will typically require
multiple probes within the littoral plot. If you have to move into shallow water to use the
sounding rod to observe sediment characteristics, use the provided comment bubble to flag
the observation and record the depth where you observed the sediment.
ฆ If the bottom is covered with materials other than mineral substrates, choose "Woody
Debris", "Organic (leaf pack, detritus)", or "Vegetation/Other".
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ฆ If the substrate is concealed and remote sampling is not possible, identify that the substrate
is "not observed" in a comment bubble associated with each zone.
Record the color of sediment within the littoral plot. Select "None" or "Other" if the sediment does not
match one of the color categories options on the Physical Habitat form.
Record sediment odor within the littoral zone. For sediment odor, the choices are "H2S" (sulfurous,
rotten egg), "Anoxic" (sewage odor), "Chemical" (strong odor like turpentine, paint, etc.), "Oil", or
"Other" (including musty, organic, and fishy odors). If "Other" is indicated, explain the observation in the
comment section of the form.
6.2.7 Estimate Aquatic Macrophyte Cover
Note and record whether macrophytes extend lake-ward from the observation point (e.g., further than
10 meters from the shoreline).
Estimate the areal cover of submerged, emergent (i.e., has erect portions above the water surface),
floating/floating leafed (either rooted or non-rooted vegetation), and total macrophytes within the
littoral zone. Cover categories range from absent to very heavy, as described in 6.2.6. Each of the three
types of aquatic macrophyte (submergent, emergent, floating) can have cover ranging from 0 - 100%.
They are evaluated independently, so the sum of their separate covers is theoretically 0% to 100% times
the number of types. So, in contrast to substrate cover percentages, the combined cover of submergent,
floating, and emergent aquatic macrophytes could theoretically add up to 300% due to the overlap of
plant types within the water column. The fourth question on the form about aquatic macrophyte cover
asks how much areal cover is there if you ignore the types and just estimate how much of the littoral
plot has aquatic macrophyte cover of any type, where this value is constrained to 0-100%.
As with substrate, estimating aquatic macrophyte cover may require multiple probes within the littoral
plot. Record all observations by filling in the appropriate bubbles on the Physical Habitat form. These
estimates can be made from the boat or while wading.
ฆ If you cannot see or probe the bottom, move closer to shore, use the provided comment
bubble to flag your observation and note your new location and depth in the comments
field on the form.
6.2.8 Estimate Fish Habitat Cover
Estimate the areal cover of potential fish habitat observed within the littoral plot and, when present, in
the separate drawdown zone plot. Littoral fish habitat cover features are within or partially within the
water and conceal fish from aquatic and terrestrial predators such as large fish, otters, kingfishers, and
osprey. When evaluating cover of potential fish habitat in the drawdown zone, however, estimate the
percentage cover that would be present at normal high water conditions, when these features would be
inundated. Cover categories range from absent to very heavy, as specified in Section 6.2.6. Record all
observations by filling in the appropriate bubbles on the Physical Habitat form. In most cases these
estimates can be made from the boat. Estimating fish habitat cover may require multiple probes within
the littoral plot. If certain cover types are not present, mark absent (0% cover) as your entry. In contrast
to the cover of substrate types, the sum of the various types of fish habitat features can exceed 100%,
sometimes summing to several hundred percent. Each type of fish cover can have cover ranging from 0
to 100%. They are evaluated independently, so the sum of their covers is theoretically 0 to 100% times
the number of cover types.
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Estimate and record cover for the following fish habitat types:
ฆ Aquatic macrophytes and Inundated Herbaceous Vegetation: Submerged, floating, or
emergent live aquatic or non-woody herbaceous plants
ฆ Woody Debris/Snags: Inundated or partially inundated dead trees, branches, or rootwads
with diameter >0.3 m (1 ft)
ฆ Woody brush/woody debris: Inundated dead or living woody vegetation <0.3 m diameter.
ฆ Inundated Live Trees: Inundated portions of trees >0.3 m in diameter
ฆ Overhanging Vegetation: <1 m from the water surface (do not include higher overhanging
vegetation, which might provide perches for birds such as kingfishers)
ฆ Ledges or Sharp Drop-offs: Overhanging banks, submerged rock shelves, and steep sloping
rock walls
ฆ Boulders: Larger than basketball size
ฆ Human Structures: Docks, barges, houseboats, swimming platforms, tires, car bodies, and
habitat enhancement structures (e.g., log rafts)
Note: In the drawdown zone you will estimate the potential fish cover. The potential fish cover
estimates are made only if there is a visible drawdown zone extending >1 m from the shoreline. For
these observations, the question is "What cover would there be if the drawdown zone were inundated -
i.e., it were to become part of the littoral zone? " Then, for example, a bunch of dried aquatic
macrophtes would be "Aquatic and Inundated Herbaceous Vegetation", as would newly-grown
terrestrial grasses. Cyprus trees left "high and dry" would qualify as "Inundated Live Trees >0.3m
diameter" and overhanging vegetation rooted above the drawdown zone could be "Overhanging
Vegetation within lm of the Surface".
6.2.9 Estimate the Cover and Type of Riparian and Drawdown Zone Vegetation
Three independent layers of riparian vegetation will be examined, each of which can range in cover from
0 to 100%. The ground layer components must add up to 100% because this layer includes bare ground
and water. The mid-layer and upper layer do not include ground or water, so vegetation within these
layers do not have to add up to 100%.
The areal cover of different types of vegetation will be estimated in the riparian plot and, when present,
in the drawdown zone. Vegetation cover is divided into three layers, which are described below.
Remember that individual plants can contribute cover to more than one layer and cumulatively the
three layers can potentially add up to 300% cover (up to 100% per layer). Also note that some things
other than vegetation are possible entries for the "Ground Cover" layer (e.g., water or barren ground).
Within each layer, the summed cover cannot exceed 100%, but, the ground layer covers must add up to
100%. As with the other visual cover estimates, you are assigning cover percentage categories, so the
summed cover is estimated by looking at various combinations of the high and low end of each
percentage cover class.
6.2.9.1 Canopy Vegetation (greater than 5 m high)
Record the type of vegetation in the canopy as deciduous, coniferous (needle-leafed evergreen),
broadleaf evergreen, or mixed, where mixed is defined as a segment where chosen if there is more than
one of these types of vegetation that has at least 10% areal coverage. If no canopy exists in the plot, do
not mark any of the bubbles.
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Estimate the areal cover of big (trunk >0.3 m diameter at breast height [dbh]) and small (trunk <0.3 m
dbh) trees. Cover categories range from absent to very heavy, as described in Section 6.2.6. Record all
observations by filling in the appropriate bubbles on the Physical Habitat form.
Total cover in the canopy layer can range anywhere from 0% to 100% depending on conditions present.
6.2.9.2 Understory Vegetation (5m to 0.5m high)
Record the type of vegetation in the understory as deciduous, coniferous, broadleaf evergreen, or
mixed, where mixed is defined as above. If no understory exists in the plot, do not mark any of the
bubbles.
Estimate the areal cover of all woody vegetation (which includes both the trunks and branches of trees
and shrubs, woody stems of perennial plants, etc.) and tall herbs, grasses, and forbs. Cover categories
range from absent to very heavy, as specified in Section 6.2.6. Record all observations by filling in the
appropriate bubbles on the Physical Habitat form. Total cover in the understory layer can range
anywhere from 0% to 100% depending on conditions present.
6.2.9.3 Ground Cover (lower than 0.5m high)
Estimate the areal cover of woody vegetation; tall herbs, grasses, and forbs; standing water or
inundated vegetation; and barren, bare dirt, or buildings. Areas of exposed rock or bedrock should be
considered 'barren'. Cover categories range from absent to very heavy, as specified in Section 6.2.6.
Record all observations by filling in the appropriate bubbles on the Physical Habitat form.
Ground cover is the only layer in which cover estimates should add up to roughly 100% (as opposed to
ranging from 0-100% in the other two layers). Certain ground cover types should be considered mutually
exclusive, i.e., if ground layer vegetation overlays barren ground, record the vegetation cover even
though there is barren ground beneath it.
6.2.9.4 Consider a tions for Dra wdo wn con ditions
Drawdown Zone vegetation entries are located to the right of Riparian Zone Vegetation on the Physical
Habitat form. They are filled out only if there is a drawdown zone extending >1 m from shore. Unlike the
case with potential fish cover, record these vegetation estimates just as you see them i.e., do not in
this case imagine that the drawdown zone is under water. For example: there must be water on the
ground (e.g., puddles) to have an entry for "standing water or inundated vegetation" in the drawdown
zone. Large trees rooted above the drawdown zone can contribute cover over the drawdown zone.
Dried aquatic macrophyte vegetation cover is entered under "Herbs, Grasses, & Forbs" with comment
that it is dried aquatic macrophytes. There may also be a lot of zeros for vegetation in the drawdown
zone (especially if the drawdown is fairly recent).
6.2.10 Record Evidence of Human influence
You will record any observations of human influences within the littoral, riparian, and, when present,
drawdown zone plots. Human influences within the littoral plot will be recorded in different locations on
the form depending on whether or not there is a drawdown plot.
When drawdown or lake level fluctuations are minimal, i.e., <1 m horizontal distance, there is no
drawdown zone plot, and the drawdown zone human influence field will be left blank. Human
influences in the littoral plot are recorded along with the influences in riparian zone plot and
recorded in the riparian zone portion of the form.
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When a drawdown zone extending >1 m from shore is present, a drawdown plot is defined and
human influences within both the drawdown and littoral zone are recorded in the drawdown
portion of the Physical Habitat form.
Within each zone, observations are recorded as not present (0), present outside and/or adjacent to (P),
or contained within (C) the plot area (Figure 6-2). Record all observations by filling in the appropriate
bubbles on the Physical Habitat form. The proximity "P" zones have a defined width of 15 m as shown in
Figure 6-2. If there is no drawdown zone plot, all C and P human disturbances in the littoral plot are
recorded on the riparian part of the form along with human influences present in the riparian zone, and
the drawdown portion of the human influences section will be left blank. In contast, if there is a
drawdown zone, all C and P human disturbances in the littoral plot are recorded in the drawdown part
of the form along with human influences present in the drawdown zone.
For each zone and influence, indicate the presence only of the influence 'closest' to the plot itself. Do
not mark "P" for a particular influence type if it is already marked "C" in that zone (use the more
influential proximity code). Human Disturbances absent (0) and within-plot (C) are straightforward. For
'Present but outside or adjacent to the plots' (P), use these guidelines:
A disturbance is marked "P" if the disturbance is seen entirely outside of any of the
plots, but is adjacent to (i.e., within 15 meters left or right hand side of the entire
Littoral-Drawdown-Riparian plot), or behind, the riparian plot within the defined areas.
A disturbance is also marked "P" if the disturbance is seen behind, but entirely outside
of all of the plots but is visible looking on-shore through the three plot zones (littoral,
drawdown, and riparian)
As a result, a single disturbance that is adjacent to both plots would be marked "P" in
both the Riparian-Littoral and the Drawdown zones.
If there is a drawdown plot, the presence of a human influence item WITHIN THE
LITTORAL PLOT is recorded as "C" in the DRAWDOWN portion of the form (e.g., consider
the littoral and drawdown zones to be a single plot when drawdown is present).
If there is NO DRAWDOWN PLOT, i.e., the riparian plot abuts the shoreline, then human
disturbances in the littoral plot are recorded by entering "C" in the Riparian portion of
the form (e.g., consider the littoral and riparian zones to be a single plot when no
drawdown is present).
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No Drawdown Zone Present
Figure 6-2 Human disturbance and proximity determinations
Cr = Marked on form as contained within riparian plot
Pr = Marked on form as adjacent to riparian plot
CD = Marked on form as contained within drawdown zone
PD = Marked on form as adjacent to drawdown zone
6.3 CyanoHABs Visual Assessment
Field crew views and documents visual observations of a potential cyanobacterial bloom in the NLA App
and photo documents the bloom according to the reporting mechanism selected by the field crew (i.e.,
bloomWatch or state-specific reporting mechanism).
6.4 Benthic Macroinvertebrate Sampling
6.4.1 Summary of Method
Benthic macroinvertebrates are collected using a semi-quantitative sampling of multiple habitats in the
littoral zone of lakes using a 500 pim mesh D-frame dip net (Figure 6-3). Sample collection is stratified on
the following specific habitat types: rocky/cobble/large woody debris; macrophyte beds; fines (including
mud, sand, or silt); and leaf packs.
Figure 6-3 D-frame net (500 |im mesh) used for collecting benthic macroinvertebrates.
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6.4.2 Equipment and Supplies
Table 6-3 provides the equipment and supplies needed for field operations to collect benthic
macroinvertebrates.
Table 6-3 Equipment and supplies - benthic macroinvertebrate collection.
Type Item Quantity
Form
NLA Littoral Samples
1
Documentation
Inner and outer labels: Benthic samples
1
Labels: Benthic extra jar
As needed
Scissors
1
Clear tape strips (to cover sample labels)
As needed
Collection
Kick net (500 nm D-shaped, modified) with 4-foot handle
1
Spare net(s) and/or spare bucket assembly for end of net
As needed
Bucket (5-gallon capacity, plastic)
1
Sieve bucket (500 nm)
1
Watchmakers' forceps
1
Squirt bottle (1 L Nalgene) - lake water
1
Spoon (stainless steel)
1
Funnel
1
HDPE bottle (1 L, white, wide-mouth)
1 or more
Ethanol (95%)
2 gal
Gloves (latex/nitrile, non-powdered, box)
2 pair
Storing and preserving
Cooler
1
Plastic electrical tape
As needed
6.4.3 Sampling Procedure
6.4.3.1 Site Selection and Sample Collection
The process for selecting the PHab stations is described in Section 6.1. All benthic samples should be
collected from the dominant habitat type within the 10 m x 15 m littoral zone component of each of the
PHab stations (Figure 3-2). The sampling process is described below. As part of the 2022 survey, new
conservation measures for native freshwater mussels were added to the benthic sampling protocol15.
6.4.3.2 Sample Processing in the Field
i/i
LU
j Use a 500-mm mesh sieve bucket placed inside a larger bucket full of lake water while sampling to carry
p the composite sample as you travel around the lake,
u
^ 6.4.3.2.1 Benthic macroinvertebrate sampling
~z.
-j 1. After locating the sample site according to procedures described in the physical habitat section,
g identify the dominant habitat type within the plot from the classifiers below:
i/i
Q
o
1=
62
_i b For this protocol, native freshwater mussels only refer to bivalves in the families Unionidae and Margaritiferidae.
q: See Appendix C for examples of freshwater mussels. The conservation measures do not apply to the invasive
quagga and zebra mussels, nor do they apply to any other mussel or clam (e.g., native fingernail clams, invasive
Asiatic clam).
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Rocky/cobble/large woody debris;
Macrophyte beds;
Fines (including mud, sand or silt); or
Leaf pack.
2. Prior to collecting the sample and when the water clarity allows, inspect the targeted substrate
for the presence of native freshwater mussels. If native freshwater mussels are present, move to
an area within the same habitat type where native mussels are absent and the previously
identified mussels will not be exposed to sampling pressures. Check the box in the NLA App that
identifies native mussels were observed and the collection location was shifted.
3. Use the D-frame dip net (equipped with 500-nm mesh) to sweep through one linear meter of
the dominant habitat type at a single location within the 10 m x 15 m littoral zone sampling
area, making sure to disturb the substrate enough to dislodge organisms.
When safe to do so, it is preferable that you exit the boat and disturb the substrate (e.g.,
overturn rocks, logs) using your feet while sweeping the net through the disturbed area.
Because a dip-net is being used for sampling, the maximum depth for sampling will be
approximately 1 m (the length of the dip-net handle); therefore, in cases in which the depth
of the lake quickly drops off, it may be necessary to sample in the nearest several meters to
the shore.
4. After completing the 1 m sweep, remove all organisms and debris from the net and place them
in a bucket following sample processing procedures described in the following section.
5. Inspect the sample for native freshwater mussels after each sweep. If one or more native
mussels were unintentionally collected:
a. In the bucket, gently rise each mussel with lake water to remove invertebrates that may
be on its shell.
b. Carefully return the native mussel to the same habitat it was sampled from and away
from further sampling pressures. They are NOT to be pushed into the substrate but to
be placed gently on their side on top of the same substrate type or as instructed by local
permit.
c. Check the box in the NLA App that native freshwater mussels were removed from the
sample and returned to the lake.
d. For EPA contractor and regional crews: when sampling a lake where ESA-listed
freshwater mussels have the potential to be present, crews must report the encounter
in the Federal ESA form in the NLA App.
e. Never intentionally collect and retain a native freshwater mussel that has the potential
to be an ESA-listed species.
6. Proceed to the next sampling station and repeat steps 1-5. The organisms and detritus collected
at each station on the lake should be combined in a single bucket to create a single composite
sample for the lake. After sampling at all PHab stations is complete, process the composite
sample in the bucket according to procedures described in the following section. One to five
bottles should be sufficient to hold the composited sample from each lake.
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If there is a large amount of debris (rocks, sticks, etc.) accumulating in the composite
sample, remove debris between sampling stations, after the debris is inspected, picked,
and/or washed to ensure no organisms are lost.
If your first collection at a sampling station results in too much debris, discard it, move the
location within the same habitat station, and take another sample.
It is recommended that crews carry a sample bottle containing a small amount of ethanol
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. Crews should NEVER,
however, attempt to 'field-pick' the samples.
6.4.3.2.2 Preparing composite samples for benthic macroinvertebrates
1. Pour the entire contents of the bucket through a sieve (or into a sieve bucket) with 500 pim
mesh size. Remove any large objects and wash off any clinging organisms back into the sieve
before discarding. Once again review the sample for native freshwater mussels and follow the
steps identified above in the sampling process.
2. Using a wash bottle filled with clean lake water, rinse all the organisms from the bucket into the
sieve. This is the composite sample for the lake.
3. Estimate the total volume of the sample in the sieve and determine how many jars (1 L jars,
each no more than half-full with sample) will be required.
4. Fill in a sample label with the Site 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 Littoral Samples 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 half 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.
6. If additional jars are needed, use a pre-printed benthos extra jar label or, if needed, fill in a blank
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
additional jars and cover them with a strip of clear tape. Record the number of jars required for
the sample on the Littoral Samples form. Make sure the number you record matches the actual
number of jars used. Write "Jar N of X" 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).
7. Place a waterproof label inside each jar with the following information written with a number 2
lead pencil:
Site ID;
Site name;
Date of collection;
Sample type;
Collector(s) names or initials; and
Sample ID.
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8. 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: Prepared composite samples
can be transported back to the vehicle before adding ethanol if necessary. In this case, fill the jar
with lake water, which is then drained using the net (or sieve) across the opening to prevent loss
of organisms, and replaced with ethanol at the vehicle.
9. 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
electrical tape.
10. 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.
6.5 Fecal Indicator Sample (Enterococci)
6.5.1 Summary of Method
Field teams are to collect a water sample within or near the littoral zone of the last PHab station where
the water is about waist deep (1 meter). The water sample should be collected when the crew first
arrives at the station (i.e., prior to substrate assessment and collection of the benthic macroinvertebrate
sample). For "large lakes" (greater than 10,000 ha) the sample is to be collected from the launch site at
the end of the day. Filters must be frozen within six hours of collection. Teams are to use a pre-
sterilized, 250 ml bottle and collect the sample at about 0.3 meter (12 inches) below the water.
Following collection, samples are placed in coolers and maintained on ice prior to filtration of two 50 mL
volumes. Again, samples must be filtered and frozen on dry ice within six hours of collection.
6.5.2 Equipment and Supplies
Table 6.4 provides the equipment and supplies needed to collect the fecal indicator sample.
Table 6-4 Equipment and supplies: fecal indicator sample
Type
Item
Form
NLA Littoral Samples
Collection
Nitrile gloves
Pre-sterilized, 250 ml sample bottle
Sodium thiosulfate tablet
Wet ice
Cooler
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6.5.3 Sampling Procedure
The procedure for collecting the fecal indicator sample is presented in Table 6-5.
Table 6-5 Procedure: fecal indicator (enterococci) sample collection
Enterococci Sample
1. Put on nitrile gloves.
2. Identify an undisturbed area within or near the littoral zone of the final habitat station where the water is
about waist deep (1 meter). For "large lakes" (greater than 10,000 ha) the sample is to be collected from
the launch site at the end of the day.
3. Lower the uncapped, inverted 250 ml sample bottle to a depth of 1 foot (0.3 m) 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. After removing the container from the water, discard a small portion of the sample to allow for proper
mixing before filtering (down to the 250 mL mark on the bottle).
6. Add the sodium thiosulfate tablet, cap, and gently shake the bottle 25 times.
Storage
7. Store the sample in a cooler on ice to chill (do not freeze immediately). Chill for at least 15 minutes before
filtering.
8. Sample must be filtered and all filters frozen within six hours of collection.
6.5.4 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 8.2 for more information regarding prcessing the Enterococci samples.
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7.0 LAKE WIDE FISH SAMPLE COLLECTION
7.1 Summary of Method
Procedures for the whole fish composite sample are described in detail in Section 7.3.The objective is to
collect one whole fish composite sample from the 636 designated lakes. This subsample is
approximately 70% of the survey design (904 lakes total). Sites with "FT" in the design panel (i.e.
Panel_Use) identify lakes designated for fish sampling. The two revisit sites in each state are also
designated as fish sampling sites. Crews should collect fish during the//'rsฃ site visit for a revisit site, to
allow for a second opportunity, if fish cannot be collected during the first visit.
The focus is on obtaining predator fish species that:
are commonly consumed by humans;
satisfy legal requirements of harvestable size for each lake site (or at least consumable size if no
legal harvest requirements exist); and
are sufficiently abundant within the lake.
Each fish sample will be a composite of five adult whole fish of the same species that are similar in size
(i.e., the smallest individual in the sample is no less than 75% of the total length of the largest
individual). See the last paragraph in 7.1 for clarification about the acceptable number of predator fish in
a composite sample.
Collection of the whole fish composite sample is a lakewide activity. It does not need to be associated
with the index site or PHab stations. Field crews may target areas in the lake with high quality habitat for
the target fish species, except for areas near a lake inlet or adjacent to riverine habitat. Crews are to
avoid these areas (a minimum of a 5-meter buffer zone) since they are more likely to contain
nonresident fish. Additionally, when sampling run-of- the river reservoirs, crews should focus their
sampling efforts in lacustrine habitats and avoid the river-reservoir transtion zone (i.e., do not sample in
habitats with flowing water). Crews should make a reasonable effort to collect a whole fish composite
sample before determining a fish sample cannot be collected. This may take up to three hours,
depending on the waterbody.
If a lake designated for fish sampling is determined target and sampleable, but the crew was unable to
collect a whole fish composite sample, no site replacement is needed. If a lake designated for fish
sampling is dropped (i.e., determined non-target or not accessible), a replacement site is identified
following procedures described in the NLA 2022 SEG and a whole fish composite sample must be
collected at the replacement site. At revisit sites, this sample is only collected at one of the two site
visits; however, crews that are unsuccessful at collecting the whole fish composite sample during visit 1
are expected to attempt the collection of that sample during visit 2. Whole fish composite samples are
shipped to the laboratory designated for interim storage of the samples.
This section contains the sampling procedures and target predator fish species for the whole fish
composite sample collection. Note that the target fish species list (Table 7-2) includes 12 primary target
predator fish species and 10 secondary predator fish species. Field crews must attempt to collect a
primary target predator fish species wherever possible. If primary target predator species are not
available at a particular site, then the field crew collects a composite of one of the secondary predator
fish species. In the event that a crew is unable to collect fish which are on either of the predator species
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lists, then the on-site biologist can select an appropriate pelagic predator. If the field crew has any
questions they should contact the NLA Fish Fillet Indicator Trainer or the EPA Technical Lead for the Fish
Fillet Contaminants Indicator (Table 2.2).
Field crews are encouraged to collect fish for the composite sample using hook and line or
electrofishing. Crews may also seine or use gill nets when this would be an efficient approach to sample
the target fish species and when allowed by the sampling permit. Crews are not to use trawling to
collect the fish. Crews may not purchase fish for the whole fish composite sample.
The whole fish composite sample should consist of five similarly sized (i.e., the total length of the
smallest specimen is no less than 75% of the total length of the largest specimen) adult predator fish of
the same species. The minimum acceptable length for a fish in any composite sample is 190 mm. Field
crews should make every effort to consistently obtain five fish for the composite sample; however, a
sample of fewer than five fish is acceptable. Conversely, for the exceptions where field crews collect
five fish that are small, they should collect up to five additional fish (for an overall composite of up to
10 fish) to provide adequate tissue for analysis. Fish retained as the fish composite sample should
remain intact and be submitted as whole specimens.
7.2 Equipment and Supplies
Table 7-1 lists the equipment and supplies necessary for field crews to collect whole fish composite
samples. A human health fish sampling kit will be provided to field crews for whole fish sampling sites
(separately from site kits) as requested by the crew (see Appendix A). Additional fish collection supplies
can be ordered through the Request form. A list of frequently asked questions and responses will be
provided with the fish sampling supplies to clarify situations that field crews may encounter while
collecting whole fish composite samples. Detailed procedures for collecting and processing the whole
fish composite samples are presented below.
Table 7-1. Equipment & supplies: whole fish composite sample collection for human health
Type Item
O
b
O
U
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LT)
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68
Form
NLA Whole Fish Sample in NLA App
Collection
scientific collection permit
electofisher, hook and line, trap nets (or other device allowed in the sample
collection permit)
sampling vessel (including boat, motor, trailer, oars, gas, and safety equipment)
nitrile gloves*
Coast Guard-approved personal floatation devices
Global Positioning System (GPS)
livewell and/or buckets
measuring board (millimeters)
Storing and
preserving
aluminum foil (solvent rinsed)*
polyethylene tubing (food-grade)*
large plastic (composite) bags*
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plastic cable ties*
human health fish sample coolers*
dry ice (for preservation) or wet ice (for temporary transport)
Documentation human health fish sample labels (individual fish labels and composite bag labels)*
fine-tipped indelible markers (for labels)
Tyvek label tag with grommet (for composite bag)*
clear tape
* Provided by EPA in FTIS kits.
**Provided by EPA in site kits.
7.3 Sampling Procedure
Note: Do not handle any food, drink, sunscreen, or insect repellant until after the composite sample has
been collectedmeasured, and wrapped (or implement measures to reduce contamination by such
chemicals if applied such as washing, wearing long gloves, etc.).
1. Put on clean nitrile gloves before handling the fish.
2. Rinse potential target species/individuals in ambient water to remove foreign material from the
external surface and place them in clean holding containers (e.g., livewells, buckets).
3. For each human health fish composite sample, select five whole fish. Criteria for inclusion in the
human health fish composite sample:
a. All fish are of the same primary target species or secondary fish species (See Table 7-2)
Note: It is essential that field crews accurately identify the organisms submitted for
analysis. Do not submit organisms from different species in a single sample.
b. All fish are adult fish; and
c. All fish 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. The minimum acceptable fish length is
190 mm.
4. Measure each fish selected for the composite 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) to
determine total body length in millimeters.
5. On the Whole Fish Sample form in the NLA App:
Verify the date of the fish collection on the top of the form. The date in this field will
be automatically entered based on the date recorded on the Verification form. If ^
you have not filled out the Verification Form or if the date of the fish collection is ^
different, you can edit the date here. Note however that changing the value on this ^
form will not change the value on the Verification Form. 0
Ensure the sample identification number is entered. ^
Q_
Check the boxes verifying that all samples are of similar length and the same ^
<
species. 1/1
Record species selected for analysis, individual specimen lengths (total length in S2
mm), and any relevant comments. Extra rows are provided in the App in the event g
that additional specimens are collected to ensure adequate tissue for analysis (refer ^
to Frequently Asked Questions for further clarification). ^
<
69
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Make sure the sample ID and specimen numbers recorded in the App match those
on the sample labels.
6. Wearing clean nitrile gloves, remove each fish selected for analysis from the clean holding
container(s). Dispatch each fish using the most humane method available.
7. Wrap each fish specimen in extra heavy-duty aluminum foil, with the dull side in contact with
the fish (foil is solvent rinsed and baked and will be provided by EPA).
8. Prepare a sample label for each sample specimen, ensuring that the label information matches
the information recorded on the Whole Fish Sample form in the App. Be sure to record the
common name and specimen length on each label.
9. Cut separate lengths of food grade tubing (provided by EPA) long enough to contain each
individual fish, allowing extra length on each end to seal with cable ties. Place each foil-wrapped
specimen into the appropriate length of tubing. Seal the ends of each tube with a plastic cable
tie. Attach the appropriate sample label to the plastic tubing by wrapping clear tape around the
label and then completely around the wrapped fish (so that the clear tape wraps over itself).
10. Double-bag the entire set of specimens in the composite by placing all fish composited from the
site inside a large plastic bag (provided by EPA). If additional bags are required for large fish
specimens or fish samples, please use plastic bags of similar thickness as those provided by EPA.
11. Fill out a Sample Identification Label for the outer bag, ensuring that the label information
matches the information recorded on the Whole Fish Sample form in the App. Be sure to record
the common name and specimen length range on the label.
12. Affix the sample label to a composite bag tag (Tyvek tag) and cover with clear plastic tape.
Thread a cable tie through the grommet in the tag and seal the outer bag with the cable tie.
SAMPLE STORAGE AND SHIPPING PREPARATION
1. After the fish sample is packaged, immediately place the fish sample in a cooler of dry ice and
use either of the following options: (1) replenish dry ice at least daily until the sample can be
properly frozen at <-20ฐC in a laboratory or other interim facility or (2) pack cooler with 50
pounds of block dry ice and ship to Microbac Laboratories (Baltimore, MD) before the end of the
day.
If a fish sample is held on dry ice in the field, the field crew should replenish the supply of
dry ice at least daily until the sample can be properly frozen or shipped.
Packaged fish samples may be placed on wet ice in coolers if they will be immediately
transported to a nearby laboratory or other interim facility to be frozen before shipment
(wet ice should be replenished frequently before it melts).
Keep all specimens in a particular fish composite sample in the same cooler for transport.
Ship only one fish composite sample in a cooler.
2. Crews have two options for freezing and shipping fish composite samples, depending on site
logistics:
a. Ship the samples via priority overnight delivery service (i.e., Federal Express), packed on
dry ice, so that they arrive at Microbac Laboratories (Baltimore, MD) within 24 hours
from the time of sample collection. Do NOT ship on Fridays, Saturdays, or the day before
federal holidays. Fish samples must be packed on sufficient dry ice (50 pounds
minimum, with blocks of dry ice layered to ensure direct contact between fish and dry
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ice) to keep them frozen for up to 48 hours. Do not use dry ice pellets for shipping the
whole fish composite sample. Remember to record the tracking number on the
Tracking form in the App before submitting it to NARS IM.
b. Add fish samples to a freezer capable of maintaining temperatures of <-20ฐC within 24
hours of collection, and store the frozen samples until shipment within two weeks of
sample collection. If fish samples cannot be stored in a freezer within 24 hours of
collection, the field crew should replenish the supply of dry ice in the cooler containing
the samples, at least daily, until the samples can be properly frozen or shipped. Frozen
fish samples will subsequently be packed on at least 50 pounds of layered blocks of dry
ice and shipped to Microbac Laboratories (Baltimore. MD) via priority overnight delivery
service. Refer to reminders in option 2a (above) about not shipping on Fridays,
Saturdays, or the day before federal holidays and about including sample tracking
numbers on App tracking forms.
Table 7-2. Primary and secondary NLA target species for human health fish collection
PRIMARY PREDATOR HUMAN HEALTH FISH TARGET SPECIES
FAMILY
SCIENTIFIC NAME
COMMON NAME
Centrarchidae
Micropterus salmoides
Largemouth Bass
Micropterus dolomieu
Smallmouth Bass
Pomoxis nigromaculatus
Black Crappie
Pomoxis annularis
White Crappie
Percidae
Sander vitreus
Walleye
Perca flavescens
Yellow Perch
Moronidae
Moron e chrysops
White Bass
Esocidae
Esox lucius
Northern Pike
Salmonidae
Salvelinus namaycush
Lake Trout
Salmo trutta
Brown Trout
Oncorhynchus mykiss
Rainbow Trout
Salvelin us fontinalis
Brook Trout
SECONDARY PREDATOR HUMAN HEALTH FISH SPECIES
FAMILY
SCIENTIFIC NAME
COMMON NAME
Centrarchidae
Lepomis macrochirus
Bluegill
Ambloplites rupestris
Rock Bass
Micropterus punctulatus
Spotted Bass
Percidae
Sander canadensis
Sauger
Moronidae
Morone saxatilis
Striped Bass
Morone americana
White Perch
Esocidae
Esox niger
Chain Pickerel
Salmonidae
Oncorhynchus clarkii
Cutthroat Trout
Coregonus clupeaformis
Lake Whitefish
Prosopium williamsoni
Mountain Whitefish
O
b
O
U
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i/i
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8.0 FINAL LAKE ACTIVITIES
Prior to leaving the lake, make a general visual assessment of the lake and its surrounding catchment.
This assessment is based on the collective observations of all crew members. The objective of the lake
assessment is to record field crew observations of catchment and lake characteristics that are useful for
future data interpretation, ecological value assessment, development of associations, and verification of
stressor data. While often subjective, these observations and impressions are extremely valuable.
In addition, review all data forms and sample labels for completeness, accuracy, and legibility. Make
sure all samples are labeled, sealed, and properly preserved. Activities described in this section are
summarized in Figure 8-1.
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Figure 8-1 Final lake activities summary
8.1 General Lake Assessment
Complete the Assessment form at the end of lake sampling, recording all observations from the lake
that were noted by all crew members during the course of the visit. This 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 comments section. The form consists of six major sections: 1)
Lake/Catchment Site Activities and Disturbances Observed, 2) General Lake Information, 3) Shoreline
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Characteristics, 4) Qualitative Macrophyte Survey, 5) Waterbody Character, and 6) Qualitative
Assessment of Environmental Values. In 2022, new questions were added to assist the determination of
natural and human-made lakes.
8.1.1 Lake/Catchment Site Activities and Disturbances Observed
Record any of the sources of potential stressors listed in Table 8-1 on the Assessment form. These
potential stressors may have been observed while on the lake, while driving or walking through the lake
catchment, or while flying over the lake and catchment. For activities and stressors that you observe,
rate their abundance or influence as low (L), moderate (M), or heavy (H) by filling in the correct bubble
next to each disturbance listed. 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
lake, fill in the "L" bubble for low next to "Houses." If the lake is ringed with houses, rate it as heavy (H).
Similarly, a small patch of clear-cut logging on a hill overlooking the lake would rate a low ranking.
Logging activity right on the lake shore, however, would get a heavy disturbance ranking. The section for
"Lake Site Activities and Disturbances Observed" includes residential, recreational, agricultural,
industrial, and lake management categories.
Table 8-1 Site activities and disturbances observed during final lake assessment.
Observe lake activities or disturbances listed and record as L (low), M (moderate), or H (heavy) intensity on the
Assessment form (except as noted below):
u
<
LU
<
74
Residences
Presence of any houses and residential buildings around the lake.
Maintained Lawns
Presence of any maintained lawns around the lake.
Construction
Presence of any recent construction in the immediate area around the lake or signs of
recent sedimentation events (depositional fans).
Pipes/Drain
Presence of any pipes or drains feeding into or out of the lake. If known, record the
type of activity with which the pipe is associated (e.g., storm sewer, plant intake) in
the "Comments" section of the form.
Dumping
Any evidence of landfill or dumping around the lake, including garbage pits and
informal dumping of large amounts of trash or cars and appliances along roads or
lakeshore. This does not include small amounts of litter. If informal dumping areas
exist, note that they are informal sites in the "Comments" section of the form.
Roads
Presence of any maintained roads in the immediate area around the lake.
Bridges/Causeways
Presence of any bridges or causeways across or in the immediate vicinity of the lake.
Sewage T reatment
Presence of sewage treatment facility.
Hiking Trails
Presence of formal hiking trails around the lake.
Parks, Campgrounds
Presence of organized public or private parks, campgrounds, beaches or other
recreational areas around the lake.
Primitive Parks, Camping
Presence of informal or primitive parks, camping areas, beaches or other recreational
areas (e.g., swimming holes) around the lake.
Resorts
Level of resort activity; this could include motels, resorts, golf courses, and stores.
Marinas
Presence of any marinas.
Trash/Litter
Relative abundance of trash or litter around the lake.
Surface Films, Scum or
Slicks
Relative abundance of surface films, scum, or slicks on the lake.
Cropland
Presence of cropland.
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Observe lake activities or disturbances listed and record as L (low), M (moderate), or H (heavy) intensity on the
Assessment form (except as noted below):
Pasture
Presence of pastures.
Livestock Use
Presence of livestock use.
Orchards
Presence of orchards.
Poultry
Presence of poultry operations.
Feedlot
Presence of feedlot or concentrated animal feeding operations.
Water Withdrawal
Any evidence of water withdrawal from the lake.
Industrial Plants
Any industrial activity (e.g., canning, chemical, pulp) around the lake or in the
catchment. Describe the type of industry in the "Comments" section of the form.
Mines/Quarries
Any evidence of mining or quarrying activity in the catchment or around the lake.
Oil/Gas Wells
Any evidence of oil or gas wells in the catchment or around the lake.
Power Plants
Presence of any power plants.
Logging
Any evidence of logging or fire removal of trees in the lake area.
Evidence of Fire
Any evidence of forest fires in the lake area.
Odors
Presence of any strong odors.
Commercial
Any commercial activity (e.g., convenient stores, shopping centers, restaurants)
around the lake or in the catchment.
Liming
Any evidence of liming activities.
Chemical Treatment
Presence of any chemical treatment facilities.
Angling Pressure
Estimate of the intensity of fishing activity in the lake.
Drinking Water
Treatment
Presence of any drinking water treatment facilities.
Macrophyte Control
Any evidence of dredging or other activities to control macrophyte growth; describe
these in the "Comments" section of the form.
Water Level Fluctuations
Any evidence of water level fluctuations due to lake management.
Fish Stocking
Any evidence of fish stocking in the lake.
Record any other oddities observed or additional information for any specific activity in the "Comments" section of
the form.
8.1.2 General Lake Information
Observations regarding the general characteristics of the lake are described in Table 8-2. Record these
observations on the Assessment form. The hydrologic lake type is a very important variable for defining
subpopulations for acidic deposition effects. Note if there are any stream outlets from the lake, even if
they are not flowing. If no lake outlets are observed, record the lake as a seepage lake. If the lake was
created by a manmade dam (not that a dam is present just to raise the water level), record the lake as a
reservoir. Otherwise record the lake as a drainage lake. Note any flight hazards that might interfere with ^
either low-altitude fly-overs by aircraft (for future aerial photography or videography) or landing on the p
lake for sampling purposes (either by float plane or helicopter). When estimating the intensity of motor >
boat usage, in addition to the actual number of boats observed on the lake during the visit, use other u
<
observations such as the presence of boat houses, docks, and idle craft. Note your opinion as to the w
swimmability of the lake in general. Observe any regular change in the lake levels and estimate the <
typical elevation change. ^
75
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Table 8-2 General lake information observed during final lake assessment.
Record general information about the lake as a whole
Hydrologic Lake
Type
Note if there are any stream outlets from the lake, even if they are not flowing. If no lake
outlets are observed, record the lake as a seepage lake. If the lake was created by a
manmade dam (not that a dam is present just to raise the water level), record the lake as a
reservoir. Otherwise record the lake as a drainage lake.
Outlet Dams
Note the presence of any dams (or other flow control structures) on the lake outlet(s).
Differentiate between artificial (man-made) structures and natural structures (beaver
dams).
Motor Boat Density
Record your impression of the density of motor boat usage on this lake (high or low). If
there is a restriction on the size of motor boat engines, check "Restricted." If motor boats
are banned, check "Banned." Consider the day of the week and weather in your
assessment as well as the number of boathouses, idle craft. Count jet skis and any other
motorized craft, which could stir up the lake, as motor boats.
Swimmability
Record a subjective impression about the aesthetics of swimming in this lake
(swimmability) along the range of "good" to "not swimmable."
Lake Level Changes
Examine the lake shoreline for evidence of lake level changes (e.g., bathtub ring). If there
are none, check "zero;" otherwise try to estimate the extent of vertical changes in lake
level from the present conditions based on other shoreline signs. Estimates should be
made to the nearest 0.1 m.
8.1.3 Shoreline Characteristics
Shoreline characteristics of interest during the final lake assessment are described in Table 8-3. Record
observations related to this portion of the assessment on the Assessment form. To estimate the extent
of major vegetation types, limit the assessment to the immediate lake shoreline (i.e., within 20 m of the
water). Also estimate the percentage of the immediate shoreline that has been developed or modified
by humans.
Table 8-3 Shoreline characteristics observed during final lake assessment.
Record percent of shoreline characteristics (within 20 meters of water):
u
<
LU
<
76
Forest
Deciduous, coniferous, or mixed forest, including sapling vegetation.
Grass
Meadows, lawns, or other open vegetation.
Shrub
Shrub vegetation
Wetland
Forested and non-forested wetlands (submerged terrestrial vegetation).
Bare Ground
Non-vegetated areas such as beaches, sandy areas, paved areas, and exposed rock.
Agriculture
Cropland, orchard, feedlot, pastureland, or other horticultural activity.
Shoreline
Modifications
Actual shoreline that has been modified by the installation of riprap, revetments, piers, or
other human modifications.
Development
Immediate shoreline area developed by human activity; include lawns, houses, stores, malls,
marinas, golf courses, or any other human-built land use.
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8.1.4 Qualitative Macrophyte Survey
Aquatic macrophytes (aquatic plants large enough to be seen without magnification) are important
indicators of lake trophic status. The most important indicator for this survey is the percentage of the
entire lake area (not just near the shore) covered with macrophytes, as perceived by observers. For both
"emergent/floating" and "submergent" coverage, choose one of the four percentage groupings (<5%, 5-
25%, 26-75%, >75%) on the Assessment form. In some cases, it will be fairly easy to estimate the
percentage from observations made at the PHab stations. In other cases, it will be an educated guess,
especially if the water is turbid or the lake is deep. After recording the areal percentage of macrophyte
coverage, record the typical density of the plants in the observed macrophyte beds as absent, sparse,
moderate, or high. Record your estimates on the Assessment form.
8.1.5 Waterbody Character
Rate the waterbody character which is the physical habitat integrity of the waterbody. Waterbody
character is largely a function of riparian and littoral habitat structure, volume change, trash, turbidity,
slicks, scums, color, and odor. The NLA 2022 attempts to define waterbody character through two
attributes: degree of human disturbance and aesthetics. Rate each of these attributes on a scale of 1 to
5. For development, give the lake a "5" if it is pristine, with no signs of any human disturbance. A "1"
would indicate that a lake is highly disturbed; for example, the entire lake is ringed with houses,
seawalls, docks, etc. For aesthetics (whether the lake is appealing or not) base the decision on any
factors about the lake that disturb you (trash, algal growth, weed abundance, overcrowding). Fill in the
bubble next to the number that best describes your opinion about how suitable the lake is for recreation
and aesthetic enjoyment today:
1. Enjoyment is nearly impossible;
2. Level of enjoyment is substantially reduced;
3. Enjoyment is slightly impaired;
4. There are very minor aesthetic problems; it is otherwise excellent for swimming, boating, and
enjoyment; or
5. It is beautiful and could not be any nicer.
8.1.6 Qualitative Assessment of Environmental Values
The primary goal of this study is to assess three major ecological values with respect to lakes: trophic
state, ecological integrity, and human health (through use of the lake). Based on your field experience,
record your own assessment of these values on the Assessment form. Write comments on these values
in this section.
1. Ecological integrity is the ability to support and maintain a balanced, integrated, adaptive
community with a biological diversity, composition, and functional organization comparable to
natural lakes of the region. Record your overall impression of the "health" of the biota in the
lake. Note any possible causes of impairment. The presence of higher order consumers (fish-
eating birds and mammals) is an indication of a healthy food web and should be noted here.
Similarly, the absence of an organism that you might expect to see is an important observation.
2. Trophic state is the rate or amount of phytoplankton and macrophytes produced or present in a
lake. Give your visual impression of the trophic status as oligotrophic (little or no biomass in the
lake water), mesotrophic (intermediate amounts of biomass in the lake water), eutrophic (large
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amounts of biomass in the lake water), or hypereutrophic (choked lake, with more biomass than
water). Give your overall impression of algal abundance and general type (e.g., filamentous). List
any observed potential nutrient sources to the lake (e.g., septic tanks and agricultural runoff).
3. Recreational value is the ability to support recreational uses such as swimming, fishing, and
boating. Record your overall impression of the lake as a site for recreation. Note any possible
causes of impairment or risk to human health. Note the presence or absence of people using the
lake for recreational activities.
Use the comments section on the Assessment form to note any other pertinent information about the
lake or its catchment. Here the field crew can record any observations that may be useful for future data
interpretation, especially data that was not captured during other sampling or data collection activities.
8.1.7 CyanoHAB assessment and report
In the Assessment form, field crews document the presence of potential cyanobacterial blooms that
were observed in areas of the lake that were not associated with the launch site, index site or physical
habitat stations (i.e., not previously reported the NLA App). Crews are to photo document the bloom
according to the reporting mechanism selected by the field crew (i.e., bloomWatch or state-specific
reporting mechanism).
If the field crew documented the potential occurrence of a cyanoHAB event at one or more station in
the lake, the Field Crew leader is responsible for reporting the bloom to the appropriate state, tribal or
local organization. See Section 2.2.4.5 for potential reporting mechanisms. A cyanoHAB report will only
be submitted when a potential bloom was observed. Reports are not needed to document the lack of a
bloom. All reports submitted via the bloomWatch app should add the project identifier 'NLA2022' in the
comment section in the app.
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8.2 Filtering: Processing the Fecal Indicator (Enterococci) and Chlorophyll-a Samples
8.2.1 Equipment and Supplies: Fecal Indicator
Table 8-4 provides the equipment and supplies needed for field crews to collect the fecal indicator
sample.
Table 8-4 Equipment and Supplies: Fecal Indicator (Enterococci) Sample
Type Item Quantity
Form
NLA Littoral Samples form
1
For recording
Fine-tipped indelible markers for filling out sample labels
1
measurements
Fecal Indicator sample labels: vial
Fecal Indicator sample labels: bag label)
2
Filter blank label if collecting filter blank
1
For processing
Nitrile gloves
1
samples
Sterile centrifuge tube (50 mL, screw top) in zip top bag
1
Filtration apparatus with collection flask
1
Sterile filter holder, Nalgene 145/147
1
Vacuum pump (electric pump may be used if available)
1
Sterile phosphate buffered saline (PBS)
1
Osmotics 47 mm polycarbonate sterile filters
1+
Sterile disposable forceps
1
Petri dishes (60 x 15, disposable)
1
Sterile microcentrifuge tubes containing sterile glass beads
2
Additional sterile microcentrifuge tube if collecting filter blank
1
Bubble bag
1
Zip-top bag
1
Dry ice
As needed
Cooler
1
8.2.2 Procedures for Processing the Fecal Indicator (Enterococci) Sample
The fecal indicator sample must be filtered before the chlorophyll-a sample since the filtering
apparatus needs to be sterile. The procedures for processing the fecal indicator sample are presented in
Table 8-5. The sample must be filtered and frozen within six hours of collection.
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Table 8-5 Procedure: Processing Fecal Indicator (Enterococci) Sample
Filtering for the fecal indicator (Enterococci) Sample
u
<
LU
<
80
Prior to beginning the filtering process, chill the Enterococci sample and PBS on WET ice for at least 15
minutes and chill the microcentrifuge tubes on DRY ice.
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, chilled phosphate buffered saline (PBS), Osmotics 47 mm
polycarbonate sterile filter box, and two sterile filter forceps.
3. Chill Filter Extraction tubes with beads on dry ice.
4. Aseptically transfer two polycarbonate filters from filter box to base of opened Petri dish. Close
filter box and set aside (this step is to prevent wind from disturbing the remaining filters in the
box but is optional in calm/controlled environments).
5. 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.
6. Load filtration funnel with sterile polycarbonate filter on support pad (shiny side up).
7. Gently shake sample collection bottle 25 times to mix well.
8. Measure 25 mL of the mixed water sample in the sterile graduated sterile 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 vacuum [3.44 psig]). Keep pumping until all liquid is in filtrate collection flask.
10. If the first 25 mL volume passes readily through the filter, measure and add another 25 mL and
continue filtration. If it was very difficult to filter the first 25 mL, proceed to step 11. 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. Pour approx. 10 mL of 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 PBS, ensuring all PBS is filtered through.
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 four 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.
15. Record the volume of sample filtered through the filter on the small yellow label (marked as
FILTER 1) and apply the label to the extraction tube (DO NOT cover with clear tape).
16. Record the volume of sample filtered through the filter on the outer bag label and apply the
label to the bubble bag (DO NOT cover with clear tape).
17. Insert tube(s) into bubble bag and zip-top bag on dry ice for preservation during transport and
shipping.
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Enterococci filter blanks will be prepared at all revisit sites during the first visit. Prepare the filter blanks
before filtering the lake sample.
1. Set up sample filtration apparatus using same procedure as used for the lake 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 (this step is to prevent wind from disturbing the remaining filters in the box but is
optional in calm/controlled environments).
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 mL of 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 vacuum [3.44 psig]). 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.
9. Record the volume of PBS filtered through the filter on the small yellow label (marked as BLANK) and
apply the label to the extraction tube (DO NOT cover with clear tape). Note that there is a specific
label for the blank sample. At sites where a blank is not collected, this label will be discarded.
10. Insert tube(s) into bubble bag and zip-top bag on dry ice for preservation during transport and
shipping.
11. Package and submit this sample to the lab with the standard samples.
12. Indicate that you have collected a filter blank by filling in the "Blank Collected" button on the Littoral
Samples form.
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8.2.3 Equipment and Supplies: Chlorophyll-o
Table 8-6 provides the equipment and supplies needed to process the chlorophyll-o sample. Much of
this equipment will have been used previously for the filtering of the Enterococci sample.
Table 8-6 Equipment and supplies - chlorophyll-a processing.
Type
Item
Quantity
Form
NLA Index Samples
1
Documentation
Labels: Chlorophyll-o sample label
1
Chlorophyll-o outer bag label
1
Clear tape strips (to cover sample labels)
As needed
Processing
Poly bottle (2 L, brown)
1
Centrifuge tube (50 mL, screw top) in zip top bag
1
Sterile disposable forceps
1
Filtration chamber (with filter holder)
1
Filtration flask (with silicone stopper and adapter)
1
Filtration pump (hand vacuum)
1
Graduated cylinder (250 mL)
1
Squirt bottle (1 L Nalgene) - de-ionized (Dl)
1
Test tube holder
1
Whatman 0.7 nm GF/F glass fiber filter
1
Storing and preserving
Cooler
1
Plastic electrical tape
As needed
Foil squares
1
Zip top bag
1
Wet ice
As needed
8.2.4 Procedures for Processing the Chlorophyll-o Samples
The procedure for processing the chlorophyll-o sample is presented below. Whenever possible, sample
processing should be done in subdued light, out of direct sunlight.
1. Place a glass fiber filter in the filter holder apparatus with the grid side down. Do not handle
the filter with bare hands; use clean forceps.
2. Gently shake the chlorophyll-o sample collection bottle to homogenize the sample, measure
and pour 250 mL of water into the filter holder using the graduated cylinder, replace the cap
of the filter holder, and pump the sample through the filter. Take care not to exceed 7
inches of Hg (approximately 3.4 psi) in the vacuum gauge on the filtration pump. If 250 mL
of lake water will not pass through the filter, discard the filter, rinse the apparatus with Dl
water, and repeat the procedures using a new filter and 100 mL of lake water. NOTE: If the
water is green or turbid, use a smaller volume to start.
3. Observe the filter for visible color. If no visible color is present, repeat step 3 until color is
visible on the filter or until a maximum of 2,000 mL have been filtered.
4. Once visible color is present and/or 2,000 mL of lake water has been filtered, record the
actual sample volume filtered on the Index Samples form and on the sample label. Rinse the
graduated cylinder and upper portion of the filtration apparatus thoroughly with Dl water to
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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.
5. Disconnect the upper portion of the filter apparatus from the lower portion. 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.
6. Place the folded filter into a 50 mL screw-top centrifuge tube and replace the 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. Seal the cap of the
centrifuge tube with plastic electrical tape
7. Record the sample volume filtered on a chlorophyll-o label and attach it to the centrifuge
tube. Ensure that all written information is complete and legible. Cover with a strip of clear
tape. Double check that the "total volume of water filtered" on the Index Samples form
matches the total volume recorded on the sample label.
8. Wrap the tube in aluminum foil and place in a zip top bag. Place the completed outer label
on the outside of the bag. Place this bag on ice in a cooler.
9. Remove the filter holder silicone stopper and adapter from the filtration flask. Pour off
water from the filtration flask.
10. Discard filter chamber, forceps and filter holder. New sterile items will be used at each site.
11. Retain the filter flask, graduated cylinder, silicone stopper, and adapter. Rinse these items
with Dl water between sites. After returning to the office or lab: thoroughly rinse the
graduated cylinder, the brown sample collection bottle and cap with tap water, complete a
final rinse with Dl water, and store for next sample event.
8.3 Preservation of Samples
Preserve the samples as specified in the specific protocol sections. Record the preservation information
on the index and littoral sample collection forms.
8.4 Preparation of Samples for Shipping
General information regarding the preparation and shipment of samples is available in Section 4.3.2.
General steps that apply to samples are the following:
ฆ Purge the Cubitainerฎ of any air bubbles, seal the cap tightly and wrap plastic electrical tape
clockwise around the cap. Place the Cubitainerฎ in a cooler with wet ice.
ฆ Seal all pertinent caps tightly.
ฆ Wrap plastic electrical tape clockwise around the caps, and then place the bottles in the cooler
with wet ice.
ฆ Keep chilled samples (water chemistry and chlorophyll-o filter) on wet ice until shipment (daily).
ฆ Keep the atrazine samples chilled on wet ice in the field and then refrigerate until shipment
(batched weekly).
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ฆ Keep the algal toxins, eDNA, and fecal indicator samples chilled on wet ice in the field and
FREEZE as soon as practicable. Keep frozen until shipping (batched weekly).
** The enterococci sample does not get tape around the cap and the sample label is not placed on
the sample bottle. This sample will be shipped on dry ice.
8.5 Data Forms and Sample Inspection
After the Assessment form is completed, the Field Crew Leader reviews all of the data forms and sample
labels for accuracy, completeness, and legibility. The other crew member inspects all sample containers
and package them in preparation for transport, storage, or shipment.
Ensure that all required data forms for the lake have been completed. Confirm that the Site ID, crew ID,
and date of visit are correct. On each form, verify that all information has been recorded accurately and
comments are used to explain data as needed. After reviewing each form, select the red circle at the
top, which will turn green after selection. This is an indication to the crew that the form has been
reviewed, but forms can be submitted before final review and amended at a later date if needed.
Ensure that all samples are labeled, all labels are completely filled in, and each label is covered with
clear tape. NOTE: Do not tape over the enterococci labels. Make sure that all sample containers are
properly sealed.
8.6 Launch Site Cleanup
Load the boat on the trailer and inspect the boat, motor, and trailer for evidence of plant material. 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 them organized
as presented in the equipment checklists (APPENDIX A: EQUIPMENT & SUPPLIES). 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. See Section 4.3 for additional information and follow appropriate state, tribal or other
applicable protocols.
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9.0 FIELD QUALITY CONTROL
Standardized training and data forms provide the foundation to help assure that data quality standards
for field sampling are met. These methods 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 NLA 2022.
9.1 Revisit Site
Approximately 10% of the target sites visited (96 lakes) will be revisited during the same field season by
the same field crew that initially sampled the lake. If a site selected for repeat sampling is dropped, then
the alternate site assigned to replace it should be revisited. The NLA 2022 Site Evaluation Guidelines
provides further information regarding the replacement of revisit sites. The primary purpose of this
"revisit" set of sites is to provide variance estimates that can be used to evaluate the survey design for
its potential to estimate status and detect trends in the target population of lakes. The time period
between the initial (Visit 1) and repeat visit (Visit 2) to a lake must be at least two weeks and should be
as long as possible.
Visit 2 will include the full set of indicators and associated parameters with two notable exceptions. A
fecal indicator blank will be collected at Visit 1 only and human health fish tissue will only be collected at
only one of the two site visits. Crews should collect fish during Visit 1 for a revisit site, to allow for a
second opportunity if fish cannot be collected during the first visit. If the crew collects fish on Visit 1,
they do not need to collect fish on Visit 2. If they are unable to collect fish on Visit 1, the crew should
collect fish on Visit 2.
9.2 Field Evaluation and Assistance Visits
No national program of accreditation for field work currently exists. For this reason, a rigorous program
of field evaluation and assistance visits has been developed to support the NLA 2022.
9.2.1 General Information
Evaluation and assistance visits will be conducted with each field crew early in the sampling and data
collection process, if possible, and corrective actions will be conducted in real time. These visits provide
both a quality 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 assistance visit also
provides the field crews with an opportunity to clarify procedures and offer suggestions for future
improvements based on their sampling experience preceding the visit. The field evaluations, while
performed by a number of different supporting collaborator agencies and participants, will be based on
the uniform training, plans, and checklists. This review and assistance task will be conducted for each
unique field crew collecting and contributing data under this program; hence no data will be recorded to
the project database that was produced by an 'unaudited' process or individual. The field evaluations
will be based on the evaluation plan and field evaluation checklist. The checklist will be made available
to all parties associated with NLA.
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One or more designated EPA, state, or contractor staff members who are qualified in the procedures of
the NLA 2022 field sampling operations will visit trained state, tribal, contractor, and EPA field sampling
crews during sampling operations on site. If membership of a field crew changes, and at least two of the
members have not been evaluated previously, the field crew must be evaluated again during sampling
operations as soon as possible to ensure that all members of the field crew understand and can perform
the procedures.
The purpose of this on-site visit will be to identify and correct deficiencies during field sampling
operations. The process will involve the following preparation steps and field day activities .Additionally,
conference calls with crews may be held approximately every two weeks to discuss issues and
clarifications as they come up throughout the sampling season.
9.2.2 Preparation Activities
1. Each Field Crew Evaluator will schedule an assistance visit with their designated crews in
consultation with the OA Officer, Regional NLA Coordinator, and respective Field Sampling Crew
Leader. Evaluators should be prepared to spend additional time in the field if needed (see
below). Ideally, each Field Crew will be evaluated within the first two weeks of beginning
sampling operations, so that procedures can be corrected or additional training provided, if
needed.
2. Each Field Crew Evaluator will ensure that field crews are aware of their visit plans and all
capacity and safety equipment will be provided for the Field Crew Evaluator.
3. Each Field Crew Evaluator will need to bring along the following in Table 9-1.
Table 9-1 Equipment and supplies - field evaluation and assistance visits.
Type
Item
Quantity
Form
Field Evaluation and Assistance Visit Checklist (sent from EPA)
1
Documentation
NLA 2022 Field Operations Manual
NLA 2022 Quality Assurance Project Plan
Clipboard
Pencils (#2, for data forms)/Pen
Field notebook (optional)
1
1
1
1
1
Gear
Field gear (e.g., protective clothing, sunscreen, insect repellent, hat, water,
food, backpack, cell phone)
As
needed
9.2.3 Field Day Activities
1. The Field Crew Evaluator will review the Field Evaluation and Assistance Visit Checklist with each
crew during the field sampling day and establish and plan and schedule for their evaluation
activities for the day.
2. The Field Crew Evaluator will view the performance of a field crew through one complete set of
sampling activities as detailed on the checklist.
Scheduling might necessitate starting the evaluation midway on the list of tasks at a site,
instead of at the beginning. In that case, the Field Crew Evaluator will follow the crew to
the next site to complete the evaluation of the first activities on the list.
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If the field crew misses or incorrectly performs a procedure, the Field Crew 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 Field Crew Evaluator is to provide additional
training and guidance so that the procedures are being performed consistently with the
FOM, all data are recorded correctly, and paperwork is properly completed at the site.
3. When the sampling operation has been completed, the Field Crew 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]; deficiencies [would
adversely affect data quality]). The Field Crew Evaluator will ensure that the field crew
understands the findings and will be able to perform the procedures properly in the future.
The Field Crew Evaluator will review the list and record responses or concerns from the
field crew, if any; on the checklist (this may happen throughout the field day).
The Field Crew Leader will sign the checklist after this review.
9.2.4 Post Field Day Activities
1. The Field Crew Evaluator will review the checklist that evening and provide a summary of
findings, including lessons learned and concerns.
If the Field Crew Evaluator finds major deficiencies in the field crew operations (e.g., less
than two members, equipment, or performance problems) the Field Crew Evaluator
must contact the EPA NLA 2022 Project Lead. The EPA NLA 2022 Project Lead will
contact the EPA Technical Lead for the Human Health Fish Fillet Contaminants Indicator
in OST and the EPA NARS QA Project Officer to determine the appropriate course of
action.
2. The Field Crew Evaluator will retain a copy of the checklist and submit to the NLA logistics lead.
3. The EPA NLA 2022 Project Lead and EPA NARS QA Project Officer or authorized designee will
review the returned Field Evaluation and Assistance Visit Checklist, note any issues, and check
off the completion of the evaluation for each field crew.
9.2.5 Summary
Table 9-2 summarizes the plan, checklist, and corrective action procedures.
Table 9-2 Summary of field evaluation and assistance visit information.
Field
The Field Crew Evaluator:
Evaluation
Arranges the field evaluation visit in consultation with the QA Officer, Regional NLA Coordinator,
Plan
and respective Field Sampling Crew Leader, ideally within the first two weeks of sampling
Observes the performance of a crew through one complete set of sampling activities
Takes note of errors the field crew makes on the checklist and immediately point these out to
correct the mistake
Reviews the results of the evaluation with the field crew before leaving the site, noting positive
practices, lessons learned, and any concerns
Field
The Field Crew Evaluator:
Evaluation
Observes all pre-sampling activities and verifies that equipment is properly calibrated and in good
Checklist
working order, and protocols are followed
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
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Corrective
Action
Procedures
Confirms that the field crew has followed NLA protocols for locating the lake and determining the
index site on the lake
Observes the index site sampling, confirming that all protocols are followed
Observes the littoral sampling and habitat characterization, confirming that all protocols are
followed
Records responses or concerns, if any, on the Field Evaluation and Assistance Checklist
If the Field Crew 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 Field Crew Evaluator finds major deficiencies in the Field Crew operations the Evaluator
must contact the EPA NLA 2022 Project Lead who, in turn, must contact the EPA Technical Lead
for the Human Health Fish Fillet Contaminants indicator if any of the major deficiencies relate to
this indicator.
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10.0 EPA Regional and Contracted Crews Only: Endangered Species Act
Conservation Measures
A generally applicable element of all NLA sampling is that field crews make every effort not to disrupt
nontarget habitats. NLA protocols are specifically designed to sample target lakes, ponds and reservoirs
and to avoid non-target habitats (e.g., rivers, marine, riparian, and wetlands). This section identifies
conservation and mitigations measures that are to be performed by EPA regional and contracted crews
(EPA crews) at all sites where federally listed species under the Endangered Species Act (ESA) are
possible, as appropriate. Before sampling or accessing an NLA site, EPA field crews will receive a list of
their sites screened for ESA-listed species possible at the sites. Crews will use this information to
familiarize themselves with the ESA species and to implement appropriate conservation measures. Any
additional ESA documentation used to supplement the EPA provided ESA information, such as contact
between the crew and local U.S. Fish and Wildlife Service office ("FWS" or "Service"), will be retained
and provided electronically to the NLA ESA lead periodically throughout the field season.
The below mitigations are not an exhaustive list of requirements for EPA crews. Crews are expected to
implement any additional conservation measures required by local permit. Crews should also be aware
of any relevant and reasonable state conservation measures, these may be completed at the discretion
of the crew lead. Field crew leads are expected to be cognizant of the federally listed species, listed
species critical habitats, and state species of concern that have the potential to occur at or near a given
sampling site, including habitats that will be used to access the sampling site. They are responsible for
making their crew members aware of potential occurrences of listed species and their critical habitat.
Efforts should be made to minimize risks to listed species and their critical habitats and avoid the take of
listed species while implementing the NLA field protocols. For sites without ESA listed species possible,
the following mitigations are not required.
10.1 Always Applicable
Crews refrain from intentionally targeting known ESA species in any way (handling, harassing,
feeding etc.).
If a known or suspected ESA-listed species is accidently encountered while sampling, it will be
returned as quickly as possible to the appropriate habitat and away from further sampling
pressures.
Any samples taken as part of the NLA survey protocol will be inspected carefully for the
presence of ESA organisms or other resources from listed taxa groups that could be associated
with collecting a non-listed species (such as glochidia on fish gills on a collected sample). Any
samples collected must be carefully inspected for their presence and, if found, removed prior to
sample preservation.
Crews will wait to allow a listed species to naturally move away from the sampling area (do not
herd or harass).
All encounters with any ESA species are to be recorded in the Federal ESA form in the NLA App,
which documents the species, number of individuals, the encounter type, and the disposition of
the individual after the encounter.
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10.2 When Entering or Leaving a Site and Shorline Activities
Be aware of the habitat requirements for each life cycle stage and avoid unnecessary activity
within that habitat. Crews are to limit damaging ESA species habitat when accessing or working
a site.
o Whenever available crews will use existing roads, pathways, or trails to the sampling site
for activities including transporting equipment and boats. Crews will adhere to a "stay
on trail" rule and refrain from roaming from designated pathways. If a trail is not
available, crews will attempt to navigate to and from the site on durable surfaces which
can tolerate repeated trampling. If no areas can be found that would tolerate repeated
trampling without notable impacts, then crews are to walk in a single file line to and
from the site whenever possible.
When walking through or at a site, avoid stepping on vegetation mounds or holes which may
contain listed reptiles, nests, or burrowing species. Whenever possible, avoid areas where listed
species are congregating.
Crews will utilize public boat docks whenever possible. Private boat docks can also be used with
owner permission.
Before putting a vessel or sampling equipment in the water, visually investigate the area for ESA
species and avoid areas where observed ESA are present.
Special Consideration: Shoreline, burrowing and sub-surface nesting ESA species possible
o Piping Plover and other listed birds which utilize ground nesting in sandy areas: in
addition to the applicable species-specific mitigations listed in FWS documentations,
crews are to avoid traversing across a beach or sandy area if such species may be
present. If an area is cordoned off, crews are not to enter a closed area.
o When walking through or at a site, avoid stepping on vegetation mounds or holes which
may contain listed reptiles, nests, or burrowing species (see below for more
information).
10.3 Index Site Activities
Listed species that may be encountered are fish, birds, mammals, invertebrates, amphibians, and
reptiles.
Prior to introducing any equipment that disturbs the water column or aquatic habitat, the area
should be carefully inspected for the presence of listed species. Avoid collecting or disturbing
listed species.
10.4 Littoral Activities
Listed species that may be encountered are fish, freshwater mussels (Unionidae and Margaritiferidae),
birds, mammals, invertebrates, amphibians, and reptiles.
Prior to introducing any equipment that disturbs the water column or aquatic habitat, the area
should be carefully inspected for the presence of listed species. Avoid collecting or disturbing
listed species.
Follow all freshwater mussel handling and documentation protocols in Section 6.4.
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10.5 Lake Wide Activites: Fish Sample Collection
10.5.1 Sites with ESA fish possible
When identifying a suspected fish ESA-listed species, each individual of the species is to be kept
in their own bucket of fresh water. The water in the bucket is to be obtained from the collection
site. The species will only be kept in the bucket for the time it takes to quickly and urgently
identify that species and to provide recovery time as needed. During identification, each
individual may not be continually handled for more than 1-minutes.
Crews must not perform active electrofishing for longer than 3 hours.
o If performing fish sampling at sites where Bull Trout (Salvelinus confluentus) are
possible, no electrofishing is allowed.
No gillnetting, trawling, or seining are allowed.
All other allowable fishing methods should not exceed 4 hours.
Crews will not perform fishing methods before sunrise or after sunset at sites where encounters
with ESA-listed species are possible.
Crews are to return any ESA-listed fish back into the lake (alive) as quickly as feasible. No ESA
fish may be kept for collection.
10.5.2 Sites with ESA freshwater mussles possible (indirect impacts to lake habitat-
based bivalves' host fishes)
Crews must not perform active electrofishing for longer than 3 hours.
No gillnetting, trawling, or seining are allowed.
All other allowable fishing methods should not exceed 4 hours.
Any collected target fish must be inspected for glochidia. If glochidia are found, the fish must be
returned alive to the site away from further sampling pressures. All fish with glochidia must be
reported in the Federal ESA form the app.
If glochidia on gills are found, crews are to be very aware of the likelihood of mussels
present and make sure to properly execute the mussel conservation methods.
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11.0 LITERATURE CITED
Baker, J.R., D.V. Peck, and D.W. Sutton (Eds.) 1997. Environmental Monitoring and Assessment Program
-Surface Waters: Field Operations Manual for Lakes. EPA/620/R-97/001. U.S. Environmental Protection
Agency, Washington, D.C.
Dodson, S.L., R.A. Lillie, and S. Will-Wolf. 2005 Land use, water chemistry, aquatic vegetation, and
zooplankton community structure of shallow lakes. Ecological Applications 15:1191-1198.
Grabarkiewicz, J. and W. Davis. 2008. An introduction to freshwater mussels as biological indicators.
EPA-260-R08-015. U.S. Environmental Protection Agency, Office of Environmental Information,
Washington, DC. Available at:
https://www.waterboards.ca.gov/water issues/programs/swamp/docs/cwt/guidance/445.pdf
Kurtz, J. C., L. E. Jackson, and W. S. Fisher. 2001. Strategies for evaluating indicators based on guidelines
from the Environmental Protection Agency's Office of Research and Development. Ecological Indicators
1:49-60.
Klemm, D. J., P. A. Lewis, F. Fulk, and J. M. Lazorchak. 1990. Macroinvertebrate Field and Laboratory
Methods for Evaluating the Biological Integrity of Surface Waters. EPA 600/4-90/030. U.S.
Environmental Protection Agency, Cincinnati, Ohio.
National Institute for Occupational Safety and Health. 1981. Occupational Health Guidelines for Chemical
Hazards (Two Volumes). NIOSH/OSHA Publication No. 81-123. U.S. Government Printing Office,
Washington, D.C. https://www.cdc.gov/niosh/docs/81-123/
Stemberger, R. S. and J. M. Lazorchak. 1994. Zooplankton assemblage responses to disturbance
gradients. Canadian Journal of Fisheries and Aquatic Sciences 51:2435-2447.
Stevens, D. L., Jr. and A. R. Olsen. 2004. Spatially balanced sampling of natural resources. Journal of the
American Statistical Association 99:262-278.
U.S. Coast Guard. A Boaters Guide to the Federal Requirements for Recreational Boats and Safety Tips.
U.S. Department of Transportation, United States Coast Guard, Washington, D.C.
https://www.uscgboating.org/images/420.PDF
USEPA. 2013. Level III ecoregions of the continental United States: Corvallis, Oregon, U.S. EPA
https://www.epa.gov/eco-research/level-iii-and-iv-ecoregions-continental-united-states
USEPA. 2000a. EPA Quality Manual for Environmental Programs 5360A1. May 2000.
http://www.epa.gov/quality/qs-docs/5360.pdf
USEPA. 2000b. EPA Order 5360.1 A2 CHG2, Policy and Program Requirements for Mandatory Agency-
wide Quality System, May 5, 2000. http://www.epa.gov/quality/qs-docs/5360-l.pdf
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APPENDIX A: EQUIPMENT & SUPPLIES
Base Kit
A Base Kit will be provided to the field crews for all sampling sites that they will go to. Some items are
sent in the base kit as extra supplies to be used as needed.
Base Kit Item Quantity Protocol
Centrifuge tube (50 mL, screw top) - extras
2
Chlorophyll-o
Centrifuge tube stand
1
Enterococci, Chlorophyll-o
Electrical tape*
2
General
F
Iter forceps (flat blade) - extras
6
Enterococci, Chlorophyll-o
F
Iters - Millipore 47mm polycarbonate 0.4 n (Box of 50)
1
Enterococci
F
Iters - Whatman 0.7 nm GF/F glass fiber (box of 100)
1
Chlorophyll-o
F
Itration chamber (with filter holder) - extras
Enterococci, Chlorophyll-o
F
Itration chamber adapter
Enterococci, Chlorophyll-o
F
Itration flask (side arm, 500 mL)
1
Enterococci, Chlorophyll-o
F
Itration pump (hand vacuum)
1
Enterococci, Chlorophyll-o
F
Itration flask stopper (silicone)
Enterococci, Chlorophyll-o
Foil squares (package of 25)*
1
Chlorophyll-o
Funnel
1
Water samples
Zooplankton
Benthics
Gloves (latex/nitrile, non-powdered, box of 100)
1
General
Graduated cylinder (250 mL)
1
Chlorophyll-o
HDPE bottle (1 L, white, wide-mouth) - extras
6
Benthics
HDPE bottle (125 mL, white, wide-mouth) - extras
6
Zooplankton
Littoral eDNA collection bottle (125 mL, clear, square) -
extras
3
Littoral eDNA collection
Integrated water sampler device (MPCA design)+
1
Water Samples
Kick net (500 nm D-shaped, modified) with 4 foot handlet
1
Benthics
Lugol's solution (250 mL bottle)
1
Phytoplankton
Metric tape measure (8 meter)
1
Secchi
Physical Habitat
Microcentrifuge tubes (extras)
5
Enterococci
Packing tape (extra rolls)*
2
General
Packing tape (on holder)*
1
General
Pail (narcotizing/concentrating chamber)
2
Zooplankton
Petri dishes (60x15, disposable)
20
Enterococci
Plankton net course (150 nm; 30 cm diameter net with a 20
cm reducing collar)
1
Zooplankton
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Base Kit Item
Quantity
Protocol
Plankton net fine (50 nm; 30 cm diameter net with a 20 cm
reducing collar)
1
Zooplankton
Plankton net carry case for 2 nets
1
Zooplankton
Plastic tranfer pipette for Lugol's
Phytoplankton
Poly bottle (2 L, brown)
1
Chlorophyll-o
Rubbermaid action packer
1
General
Secchi disk (20 cm diameter) with weightt
1
Secchi
Sieve bucket (500 nm)+
1
Benthics
Small tote with lid
1
General
Sodium Thiosulfate (25 tablets in vial)
1
Enterococci
Sounding line (50 m, marked in 0.5 m intervals) with clip
1
Depth
Secchi
Zooplankton
Sounding rod (3 m , marked in 0.1 m increments, PVC, 2-
section)+
1
Index Site Depth
Physical Habitat
Sounding weight with clip
1
Depth
Spoon (stainless steel)
1
Benthics
Squirt bottle (1 L Nalgene) - for de-ionized (Dl) water
1
General
Squirt bottle (1 L Nalgene) - for lake water
1
General
Surveyor's tape (50m)+
1
Depth
Physical Habitat
Tape strips (3M, pad of 25)*
General
Watchmaker's forceps
1
Benthics
* Items may need to be replenished by field crews during field season
+ Item supplied if needed
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Site Kit
A Site Kit will be provided upon request to the field crews for each sampling site. Please submit an
electronic Request form well in advance of field sampling to request the Site Kits. Each site kit will also
include necessary coolers and shipping supplies for all samples collected.
Quantity
Site Kit Item
Per Site Kit
Protocol(s)
Centrifuge tube (50 mL, screw top) in bag
1
Chlorophyll-o
C02 (Alka seltzer) tablets (packet of 2)
1
Zooplankton
Cooler liners (1 per cooler)
-
General
Cubitainerฎ (4L)
1
Water Chemistry
FedEx Express shipping labels (1 per cooler)
-
Sample shipping
Gloves (latex/nitrile, non-powdered)
4 pair
Sample collection
Filtration unit (sterile 250 mL filter funnel, cap, and filter
Enterococci
holder)
1
Chlorophyll-o
Filter forceps (sterile, flat blade)
2
Enterococci
Chlorophyll-a
HDPE bottle (1 L, white, narrow mouth)
1
Phytoplankton
HDPE bottle (60 mL, white, wide-mouth)
1
Atrazine
HDPE bottle (125 mL, white, wide-mouth)
2
Zooplankton
HDPE bottle (1 L, white, wide-mouth)
2
Benthics
Littoral eDNA collection bottle (125 mL, clear, square)
1
Littoral eDNA collection
Enterococci bottle (250 mL, sterile, clear, narrow-mouth)
1
Enterococci collection
HDPE bottle (500 mL, white, wide-mouth)
1
Algal Toxins (MICZ)
eDNA bottle (1 L, clear, square, narrow-mouth)
2
eDNA (Index and Littoral)
Phosphate buffered saline (sterile PBS)
1
Enterococci
Microcentrifuge tubes w/glass beads (in bubble & Zip bags)
2
Enterococci
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Human Health Fish Sampling Kit
A human health fish sampling kit will be provided to the field crews for designated human
health fish sampling sites (separately from site kits). Please submit an electronic Request form
well in advance of field sampling. Kits must be requested at least two weeks before sampling is
to take place. Prior to sampling, inspect each site kit to ensure all supplies are included. These
human health fish sampling kits include:
Human Health Fish Sampling Kit Item
Quantity
Aluminum foil (solvent-rinsed and oven dried)
5 packs
Cable ties
24
Cooler
1
Dry ice Shipping Label
1
FedEx airbill (Pre-addressed)
1
Frequently Asked Questions handout
1
Heavy-duty food grade polyethylene tubing
1 roll
Large plastic composite bag
1
Nitrile gloves
10
NLA 2022 Human Health Fish Fillet Contaminants Site List
1
Tyvek tag
1
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Electronic Forms & Labels
Apple iPads Labels will be supplied by the NARS IM Center upon request.
Item Quantity Protocol
Apple iPads:
2
General
Field forms packet (paper backup copies as needed):
1
General
NLA 2022 Verification
NLA 2022 Profile Calibration
NLA 2022 Profile Data
NLA 2022 Index Samples
NLA 2022 Littoral Samples
NLA 2022 Physical Habitat
NLA 2022 Assessment
NLA 2022 Tracking
NLA 2022 Whole Fish Sample
NLA 2022 Federal ESA
Labels packet (for samples)
1
General
Field Crew Supplied Equipment
This equipment will need to be supplied by the field crew. Some items are optional.
Field Crew Supplied Item Quantity Protocol(s)
Access instructions
1
Site Evaluation
Access permission documents/permit (if required)
1
Site Evaluation
Barometer or elevation chart to use for calibration
1
Calibration
Binoculars
1
Physical Habitat
Bleach (or bleach alternative)
1
General
Buckets (5 gallon capacity, plastic)
Benthic Macroinvertebrates
Buoy (for marking observation point)
1
Physical Habitat
Calibration cups and standards (for multi-parameter meter)
1
Calibration
Calibration QC check solution (for multi parameter meter,
pH and conductivity)
1
Calibration
Clinometer
1
Physical Habitat
Clipboard
1
General
Depth Finder (hand-held or boat mounted sonar)
1
Index Site Profile
Physical Habitat
Electronic data capture devices (tablet/phone/computer)
with NARS App and extra battery pack (if needed)
1-2 (optional)
General
Ethanol (95%)
Benthic Macroinvertebrates
Zooplankton
Field gear (e.g., protective clothing, sunscreen, insect
repellent, hat, water, food, backpack, cell phone)
General
Field notebook - optional
1
General
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Field Crew Supplied Item
Quantity
Protocol(s)
Field thermometer (not mercury)
1
Calibration
Fishing gear (e.g., electrofishing unit, fishing rods, nets,
livewell and/or buckets, measuring board etc.)
Whole Fish Sample
Lake Verification
GPS unit (with manual, reference card, extra battery)
1
Index Site Coordinates
Physical Habitat
Kick net (500 nm D-shaped) with 4 foot handle (spare)
1
Benthic Macroinvertebrates
Laser rangefinder (for estimating drawdown) - optional
1
Physical Habitat
Map wheel or string (for measuring shoreline distances on
site map)
1
Physical Habitat
Multi-parameter water quality meter (with temperature,
pH, and DO probes)
1
Index Site Profile
Multi-parameter communication cable (50 m)
1
Index Site Profile
Net(s) and/or bucket assembly for end of net (spares)
1
Zooplankton
NLA 2022 Fact Sheets
20
General
Pen
1
General
Pencils (#2, for data forms)
2
General
Permanent marker (fine tip, for labels)
2
General
Scissors
1
General
Shipping tape
1
Shipping
Surveyors rod - optional
1
Physical Habitat
Water (deionized)
General
Boat Equipment List
This is suggested boat equipment.
Item
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Anchor (with 75 m line or sufficient to anchor in 50 m depth)
Boat horn
Boat plug (extra)
Bow/stern lights
Emergency tool kit
Fire extinguisher
First aid kit
Gas Can
Hand bilge pump
Life jackets
Motor
Oars or paddles
Second anchor for windy conditions and littoral sampling (w/ 75m line)
Sonar unit
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| Spare prop shear pin
| Type IV PFD (throwable life saving device)
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APPENDIX B: SHIPPING GUIDELINES
General Shipping Guidelines
Samples will be shipped by the field crews according to the chart below. The Field Crew Leader will
complete the appropriate section(s) of the Tracking form for the samples and will submit tracking via
the NLA App. The Field Crew Leader will place the samples and the packing slip (in a waterproof bag or
plastic sleeve) in the shipment cooler, and then the Field Crew Leader will attach the appropriate pre-
addressed FedEx label from the site kit or cooler marked for the appropriate lab. The field crew will
either drop off the cooler for shipment at a local FedEx location or arrange for a pick up at the hotel or
other appropriate facility. If the field crew has chosen a pick up, they must follow up with the facility at
which it has been left and/or track the package through FedEx tracking tools to ensure that the
shipment cooler has actually been picked up by FedEx. Once the package is in the possession of FedEx,
the IM Team and Field Logistics Coordinator (FLC) will track the package to its destination and take steps
necessary to ensure timely delivery.
A packing slip must be filled out to accompany each sample shipment. Be very careful to fill in the
information correctly and legibly, especially the Site ID. When using a packing slip that was paired with a
sample label sheet, the Sample ID numbers will be pre-printed on the packing slips. The packing slip is to
be placed in a resealable plastic bag/pouch secured to the inside of the cooler lid. Seal the shipping
container with clear packing tape. In the NLA App, fill in the shipping details in the pertinent section(s) of
the Tracking form and submit the Tracking form to the NARS IM Center to indicate that samples will be
in transit to the laboratory. The Tracking form must be submitted the same day that the samples are
shipped.
Before shipping, it is very important to preserve each sample as directed in the sample collection
portion of the appropriate chapter in the NLA 2022 FOM. General directions for sample processing,
shipping and tracking are found below:
Preserve the samples as specified for each indicator before shipping.
Be aware of the holding times for each type of sample.
Always line the cooler with a heavy duty plastic bag (cooler liner) when shipping preserved
samples or when shipping wet ice.
When shipping frozen batched samples, use the foil-backed dry ice liner and foam pad inside the
cooler.
When shipping with dry ice, it's best to layer or intermix dry ice and samples to increase the
contact between samples and dry ice.
When shipping samples preserved with ethanol, surround the jars with crumpled newspaper or
other absorbent material.
When wet ice is used for shipment:
Ensure that the ice is fresh before shipment and use adequate amounts of ice to ensure samples
will remain cold for up to 48 hours.
Place samples and all ice inside the cooler liner and seal the cooler liner to avoid leakage of
melted ice.
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Secure the cooler with clear packing tape.
When dry ice is used for shipment:
Note: Not all FedEx locations will accept shipments containing dry ice. Dry ice shipments can be shipped
from "FedEx staffed" locations, which is how these locations are referred to online. You can also arrange
for a pick-up from your lab or hotel. Dry ice shipments usually cannot be shipped from FedEx Officeฎ
locations, FedEx Retail locations such as Walgreens/Walmart/OfficeMax, or at FedEx Authorized
ShipCenterฎ locations. These types of locations are differentiated on FedEx.com in the "Find FedEx
Locations" feature. Please be sure to call in advance to ensure your location will accept the package for
shipment.
Label the cooler with a Class 9 Dangerous Goods label
o Place the label on the front side of the cooler, not the top.
o If it is not already completed, fill out the upper corners of the label with the shipper and
recipient information as on the FedEx airbill,
o Declare the weight (in kg) of the dry ice in the lower right hand corner of the label,
ensuring it is the same weight listed on the airbill.
Ensure that the dry ice is fresh before shipment, and use adequate amounts of dry ice to ensure
samples will remain frozen for up to 48 hours.
Secure the cooler with clear packing tape. Do not completely seal the entire edge of the cooler
such that the pressure inside the cooler could build.
Place the provided FedEx airbill on the top of the cooler or on a handle tag secured to one of the
cooler's handles.
o Ensure the label indicates the amount of dry ice in the package and matches the weight
listed on the dry ice label.
Tracking Forms and Shipment Types
Whenever NLA samples are shipped, one or more sections of the Tracking form is completed to relay
important shipping information to the IM Team, FLC and the destination lab. The Tracking form is
submitted electronically at the time of shipping and a packing slip is included as a hard copy in the
shipping container with the samples.
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Each section for the Tracking form has been assigned a "T" number to help crews identify the correct ^-j
section of the form to use when sending samples. This "T" number is located on the top of each tracking 9
form section). Crews will also find reference to the same "T" numbers on the individual sample labels ej
and on the top of the pre-printed FedEx return labels provided in the site kits or with batch coolers. The ^
FedEx return labels are pre-paid and allow crews to ship samples to any of the nationally contracted ^
laboratories. States using their own labs for certain samples will need to arrange for shipping on their
own.
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When crews order site kits (via the Request form), a set of packing slips will accompany each set of
labels in the site kit. Sample IDs for the suite of samples collected at a single site will be pre-populated ฃ
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on both the labels and the packing slips. By entering the water chemistry sample ID in the Tracking
form, the rest of the sample IDs will auto-populate in the Tracking form. It is important to keep the
labels and packing slips organized so the sample IDs will match when shipping occurs.
Crews include the pertinent packing(s) slip in the cooler (placed in the plastic sleeve affixed to the inside
of the cooler lid) when they send samples to the labs, and they also must submit the completed section
of the Tracking form that is associated with that sample shipment via the NLA App. If a cooler contains
samples from more than one site, then multiple packing slips must be placed in the cooler and the
Tracking form for each site must be submitted.
When a crew visits a site with the intent to sample, they complete and submit the Verification form via
the NLA App. It is very important to submit this form as soon as possible after every attempted sampling
event to provide key items such as the date of the event, the crew ID, and whether the site wa
ssampled. Prompt status reports allow the FLC to closely track sampling progress. More importantly, it
enables NARS IM to track samples that were collected at each successfully sampled site versus those
that were not, and to immediately track the shipment of the time-sensitive samples after each sampling
event. Submitting the Verification form is crucial to report the date of a completed sampling event and
must be submitted (along with sample tracking information) on the same day that samples are shipped.
Procedure for filling out and submitting tracking via the App
1. After ensuring all of the samples to be shipped are properly preserved and prepared for
shipment, access the Tracking form in the App.
2. Ensure the correct water chemistry sample ID has been entered at the top of the form. Doing so
will populate the sample IDs of all other collected samples. Samples that were not collected will
display a blank sample ID field and the not collected bubble will be transferred from the
individual sample collection forms. The not collected bubbles are not editable in the Tracking
form; to change the collection status of a sample, access the pertinent sample collection forms
(e.g., Index Samples, Littoral Samples, and/or Whole Fish Sample forms).
3. In the pertinent section of the Tracking form , check the box under the 'To Ship' column for each
sample being sent in the shipment.
4. Click the 'Enter Shipping Details' button and fill out the resulting popup window with the
destination lab, date shipped, tracking number, sender and sender's phone number.
5. Click the 'save shipping info' button to save the details and the Tracking form .
6. Once the shipping details have been saved in the App, a date will appear in the shipped column
of the Tracking form . If the shipping details for a sample need to be edited, click the date in the
shipped column to access the saved shipping details. Editing the details in this manner changes
ONLY one sample at a time. The only way to enter shipping details for an entire group of
samples is during the initial details entry.
a. If the status of the sample needs to change from shipped to not shipped, click the date
in the shipped column to access the saved shipping details and delete all the shipping
info. Click the "save shipping info" button after deleting all the shipping information.
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The sample will no longer be marked as shipped and the "to ship" checkbox will
reappear.
7. After all pertinent shipping details have been saved, click the SUBMIT menu button and select
the button next to 'Tracking' and any other the forms that you wish to submit. Click the green
submit button at the bottom of the form list. An email will pop up on your device addressed to
NARSFieldData@epa.gov. Copy yourself, any other crew members or managers and click send.
To ensure that he email was sent, check the SENT mailbox on your email app and look for the
recent email containing the data. If the email is not in the SENT mailbox, it was not sent and you
should try again after verifying an internet connection.
8. At any point, if it is determined that data needs to be revised or updated, crews should feel free
to do so in the App and re-submit any edited data or tracking forms using the steps above.
Newly revised data will automatically take the place of previous data. It is not necessary to re-
submit data or tracking forms that were unchanged however.
9. After submission, a data summary will be automatically emailed back to the email address from
which the submission was received. The Field Crew Leader or his/her designee should review
this data summary for accuracy and make any corrections necessary and re-submit the pertinent
form(s).
Shipping Groups:
T-l - Daily Water Chemistry Samples
Complete the T-l section of the Tracking form for the samples that are shipped immediately
after each sampling event (e.g., same day as sampling or the next day):
o water chemistry
o chlorophyll-o
These samples are shipped together in the site kit cooler with a heavy duty liner bag and ample
wet ice.
Samples from two sites may be shipped together in a single cooler if they were collected on the
same day.
Send the Tracking form and all data forms from the site to the IM Team via the NLA App.
Ship all of the samples to the lab in the same cooler with the packing slip that was provided with
the label packet. If any sample listed on the packing slip is not being shipped, line out the sample
ID to indicate that the sample is not in the cooler. If samples from multiple sites are shipped
together, then multiple packing slips must be used.
Samples need to be shipped on as much fresh wet ice as will fit in the cooler liner. Water
chemistry and chlorophyll-o^ samples should be shipped within 24 hours of collection (i.e., the
same day as sampling or the following day).
T-2 - Frozen Batched Samples
Complete this section of the Tracking form when shipping the frozen batched samples:
o Enterococci (frozen)
o eDNA (index and littoral; frozen)
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o algal toxins (frozen)
o atrazine (chilled)
These samples are shipped together in a cooler (ordered via the Request form) with a two-piece
dry ice liner and 20 pounds of dry ice.
2-3 site's worth of samples may be shipped together in a single cooler.
Ship all of the samples to the lab in the same cooler with the packing slip that was provided with
the label packet. If any sample listed on the packing slip is not being shipped, line out the sample
ID to indicate that the sample is not in the cooler. If samples from multiple sites are shipped
together, then multiple packing slips must be used.
To pack the T2 cooler:
1. Place the foam pad in the bottom of the T2 cooler and line the cooler with the foil-
backed dry ice liner.
2. Place frozen samples from up to three sites and dry ice (approximately 20 lbs) inside
the dry ice liner, intermixing samples and dry ice.
3. Please the enterococci samples on top of the frozen bottles to prevent damage during
shipping.
4. Place the foil-backed separator pad on top of the frozen samples and dry ice creating
a thermal barrier within the cooler.
5. Place the chilled atrazine samples (up to 3) inside the atrazine sleeve and insert the
foam plug into the end of the sleeve.
6. Place the sleeve ON TOP of the second foam pad inside the dry ice liner (such that the
foam pad is used as a separator between the dry ice and the atrazine samples).
7. Close the "lid" portion of the dry ice liner.
8. Insert the T2 packing slip(s) into the provided pouch inside the cooler lid.
9. Close the cooler lid, taping and applying FedEx shipping label as usual.
Frozen batched samples should be shipped at least every week.
In cases where a state lab is processing some, but not all of the samples, first select "To Ship"
checkboxes for the samples being sent to the national lab and select the appropriate lab in the
shipping details popup window. The T-2 packing slip should be placed in the cooler being
shipped to the national lab and should only indicate the sample(s) being shipped in that cooler.
Line out the sample IDs to indicate which samples are not in the cooler. Select "To Ship"
checkboxes for the remaining samples being sent to the state lab and select the appropriate lab
in the shipping details popup window.
Verify the sample IDs for the sample(s) being shipped.
Submit the Tracking form to NARS IM in the NLA App using the steps above.
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T-3 - Non-Chilled Batched Samples
Complete this section of the Tracking form for shipping batches of non-chilled samples:
o Benthic macroinvertebrates
o Zooplankton (course and fine)
o Phytoplankton
These samples are shipped together in a cooler (ordered via the Request form) with a heavy
duty liner bag and no ice.
The number of samples that will fit in a cooler will depend largely on the number of benthos
bottles collected from each site. In most cases, samples from three to six site visits will fit in a
cooler together. NEVER split samples from one site into more than one cooler.
Ship all of the samples to the lab in the same cooler with the packing slip that was provided with
the label packet. If samples from multiple sites are shipped together, then multiple packing slips
must be used.
Place 1-liter benthos bottles upright in the lined cooler and use newspaper or cardboard to fill
any empty space between benthos bottles to keep them in an upright position during shipping.
Place bagged zooplankton bottles on top of the upright benthos bottles.
Non-chilled batched shipped samples should be shipped within two weeks of collection.
In cases where a state lab is processing some, but not all of the samples, first select "To Ship"
checkboxes for the samples being sent to the national lab and select the appropriate lab in the
shipping details popup window. The T-3 packing slip should be placed in the cooler being
shipped to the national lab and should only indicate the sample(s) being shipped in that cooler.
Line out the sample IDs to indicate which samples are not in the cooler. Select "To Ship"
checkboxes for the remaining samples being sent to the state lab and select the appropriate lab
in the shipping details popup window.
Verify the sample IDs for the sample(s) being shipped.
Submit the Tracking form to NARS IM in the NLA App using the steps above.
NOTE: Federal regulations and FedEx rules allow for ground shipping of certain quantities of flammable
liquids WITHOUT the need for special certifications and labeling. Flammable liquids may NOT be shipped
via air carrier unless shipper is trained and qualified to do so and specific documentation and labeling
requirements are met.
The Code of Federal Regulations (49 CFR Section 173.150) lists the exceptions which allow shipping of
flammable liquids via ground carrier without labeling or special certifications. Ethanol and formalin can
be considered to be in either Packaging Group 2 or 3, so we use the more stringent PG 2 as our guideline.
The limited quantity exclusion allows ground shipping of PG 2 flammable liquids provided that the
individual containers inside the package are not over 1.0 liters each, that the gross weight of the package
does not exceed 66 pounds, and that the outer packaging is a sturdy container. Please ensure that your
shipment meets these criteria to ensure the legal ground shipment of these samples.
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1T4 - Whole Fish Composite Sample - designated sites only
Complete this section of the App tracking form for shipping frozen whole fish tissue samples
(FTIS).
Only one fish composite sample may be shipped in a single cooler.
Ship the sample to the lab in the whole fish tissue kit cooler with the packing slip that was
provided with the label packet.
Samples need to be shipped with a minimum of 50 pounds of dry ice (blocks of dry ice only).
Human health fish composite samples should be shipped within 1 week of collection.
Verify the sample ID for the sample being shipped.
Submit the Tracking form to NARS IM in the NLA App using the steps above.
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Shipping Addresses
USEPA Laboratory, Corvallis, Oregon (Water Chemistry, Chlorophyll-o)
Attn: Judy Greydanus, CSS
c/o U.S. EPA
Willamette Research Station
3080 SE Clearwater Dr.
Corvallis, OR 97333
Microbac Laboratories, Inc. (Whole Fish Composite Samples)
Attn: Sample Receiving
2101 Van Deman Street
Baltimore, MD 21224
Great Lakes Environmental Center, Inc. (All other samples)
Attn: Mike Dunlop
739 Hastings Street
Traverse City, Ml 49686
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Table B-l. Sample preservation, packaging, and holding times
Sample
Group & Lab
Sample Type
Sample
Code
Location
Sample
Target
Volume
Container
Preparation/
Preservation
ShippingTime
Frame
Packaging for
Shipping
T-l-Daily
Water Chemistry
Samples
WRS Laboratory
- Corvallis, OR
Water
chemistry
[raw,
unfiltered site
water)
CHEM
Index
4 L
Cubitainer
(4 L)
Wet ice in field
Immediate
Water Chemistry
Index
Collection
2 L
Poly bottle (2 L, brown)
Wet ice in field
(ship within 24
hours of
sampling)
Cooler with wet ice
PRIORITY
OVERNIGHT
Chlorophyll-a
CHLX
Processing
Stain on
filter -
max 2 L
filtration
centrifuge tube (50 mL),
in zip-top bag
Wet ice in field
(after filtration)
Fecal indicator
[Enter ococci)
ENTE
Last habitat
station *
200 mL
Sterile 250 mL bottle
(clear, square)
Dry Ice
T2 - Frozen
Batched Samples
eDNA
FDNA
Index
1 L
Clear square bottle (1 L)
Wet ice in field,
freeze ASAP
Frozen
Batched Cooler with
LDNA
Littoral Stations
(10) *
1 L
Clear square bottle (1 L)
Wet ice in field,
freeze ASAP
Batch up to
one week
maximum
2-piece dry ice liner
and 20 pounds dry
GLEC - Traverse
City, Ml
Algal toxins
MICZ
Index
500 mL
HDPE bottle (500 mL,
white, wide-mouth)
Wet ice in field,
freeze ASAP
ice
PRIORITY
OVERNIGHT
Atrazine
TRIA
Index
50 mL
HDPE bottle (60 mL,
white, wide-mouth)
Wet ice in field,
keep chilled
Place chilled in
provided sleeve
Phytoplankton
PHYX
Index
1 L
HDPE bottle (1 L, white
narrow mouth)
Lugol's added in
Field
Wet ice in field
T3 - Non-Chilled
Batched Samples
GLEC - Traverse
Zooplankton
[coarse -150
Hm)
(fine - 50 |im
ZOCN
ZOFN
Index
Vertical
tow(s) 5-
meter
total
length
HDPE bottle (125 mL,
white, wide-mouth)
95% ethanol
added in field
Batch up to
two weeks
maximum
Non-Chilled
Batched Cooler with
absorbent material
No Ice
GROUND
City, Ml
Benthic
invertebrates
BENT
Littoral Stations
(10+)*
All
organisms
in grabs
HDPE bottle (1 L, white,
wide-mouth)
95% ethanol
added in field (at
east 500 mL per
bottle)
T4-Whole Fish
Composite
Sample
Microbac
Laboratories,
Inc. - Baltimore,
MD
Whole fish
FTIS
Lakewide
5-10
whole fish
(minimum
fish
length
190 mm)
Wrapped individually ins
olvent rinsed foil
Sealed in poly tubing
Large outer plastic bag
Dry ice in field,
hold in freezer
Ship weekly
(except on
Fridays,
Saturdays, or
the day before
Federal
holidays)
to FTISJab.
Only ship one
sample per
cooler.
Ship in
provided FTIS cooler
with 50 pounds of
DRY ICE
PRIORITY
OVERNIGHT
* At large lakes (lakes over 10,000 hectares) littoral sampling is encouraged but not required. If littoral stations are not
visited, BENT will NOT be collected; ENTE and LDNA will be collected at the launch site; LDNA will then be a single
1L grab sample
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Field Operations Manual
Page 110 of 116
antero-
dorsal
anterior
antero-
ventral
Up to 1.5in (3.8 cm)in Inegth
Up to 5in (12.7cm) in length
Up to 2.8in (7.1 cm) in length
c Most photos, unless stated otherwise, were obtained through the ECOS website maintained by the U.S. Fish and Wildlife Service.
d Photo by G. Thomas Watters, Ohio State University
APPENDIX C: NATIVE FRESHWATER MUSSEL EXAMPLES
Figure C-l. Bivalve anatomy and orientation (Grabarkiewicz, J. and W. Davis 2008).
Narrow Pigtoe (Fusconaia escambia)' Pink Mucket (Lampsilis abrupta)
Up to 3in (7.5cm) in length
Rayed Bean (Villosafabalis)
Up to 4.1in (10.5cm) in length
Sheepnose (Plethobasus cyphyus) Snuffbox (Epioblasma triquetral
postero- dorsal
dorsal
posterior
postero-
ventral ventral
pseudocardinal
/ teeth
lateral teeth
anterior
adductor
scar
posterior ^
adductor scar
sulcus
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APPENDIX D: NLA 2022 HANDPICKED SITES: RESAMPLING OF THE
NATIONAL EUTROPHICATION STUDY LAKES
Background
In 1972, the US Congress passed the landmark environmental legislation that became known as the
Clean Water Act (CWA). The CWA has become a model for countries around the world and is heralded
as the gold standard of environmental legislation. This year, 2022, marks the 50th Anniversary of the
CWA, which coincides with the 4th iteration of the National Lakes Assessment (NLA). One hallmark of
the CWA was inclusion of monitoring and measuring progress as an integral component to restoration
and protection efforts. To that effect, the U.S. initiated a survey in 1972 to measure and report on the
acceleration of eutrophication in lakes by assessing nutrient pollution known as the National
Eutrophication Survey (NES). Importantly, the year 2022 also represents the 50th Anniversary of this
landmark national survey.
Today, the NARS program is an EPA, State and Tribal partnership designed to address the need for
monitoring the long-term progress toward the water quality objective and goals of the CWA. Through
NARS, EPA and our partners are providing information on the condition and trends in coastal waters,
lakes, rivers and streams, and wetlands. As a part of the NLA starting in 2007, ~200 of the original ~800
NES lakes were included as a subpopulation in the survey design and resampled. These ~200 lakes were
selected using a probability-based design, similar to the broader NLA, which allowed change estimates
to be made for the broader population of ~800 NES lakes. This ~200 lake subpopulation was used to
report on the changes in trophic condition in the 2007 report.
As part of the celebration of the 50th Anniversary of the CWA, a resurvey of the NES lakes is taking place
in conjunction with the NLA 2022. The statistically valid sampling design for NES sampling since the NLA
2007 survey will allow EPA to track environmental progress by assessing the change in condition of the
NES lakes in the intervening 50 years (1972, 2007, 2012, 2017, and 2022). In additions to reporting on
the condition of the NES lakes, the NES 2022 resampling will combine field data with watershed
information to identify the progress we have made, the challenges we still face, and highlight the
importance of water quality in our lives.
Field Crews and Coordination
The NES 2022 will rely on many of the key contacts and coordinators for NLA 2022 (See Table 2-2). All
ten EPA Regions are supporting the NES field work in addition to the NLA field sampling contractors. In
some regions, the field crew leads are the same as the regional NLA coordinator (Table D-l).
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Table D-l NES EPA Regional Crew Contact information
Title Name Contact Information
Regional EPA NLA Hilary Snook*, Region 1
Coordinators
snook.hilarv@eoa.gov
617-918-8670
Emily Nering*, Region 2
nering.emilv@eoa.gov
732-321-6764
Frank Borsuk*, Region 3
Leah Ettema, Region 3
borsuk.frank@eoa.gov
304-234-0241
ettema.leah@eoa.gov
304-234-0245
Chris McArthur, Region 4
mcarthur.christooherPeoa.gov
404-562-9391
Mari Nord*, Region 5
nord.mari@eoa.gov
312-886-3017
Rob Cook*, Region 6
cook, robert (Seoa.gov
214-665-7141
Gary Welker*, Region 7
welker.garv@eoa.gov
913-551-7177
Liz Rogers, Region 8
Tom Johnson, Region 8
Rogers.liz@epa.gov
303-312-6974
Johnson. tom@ eoa.gov
303-312-6226
Tina Yin, Region 9
Matthew Bolt, Region 9
yin.christina@eoa.gov
415-972-3579
Bolt.matthew@eoa.gov
415-972-3578
Lil Herger*, Region 10
herger.lillian@eoa.gov
206-553-1074
Regional EPA Field Crew Leads and/or Alternate Leads
Tom Faber, Region 1
Faber.tom@eoa.gov
617-918-8672
Raffaela Marano, Region 3
Marano.raffaela@eoa.gov
215-814-2397
Jerry Ackerman, Region 4
ackerman.ierry@eoa.gov
706-355-8721
Sue Dye, Region 4
dve.sue@eoa.gov
706-355-8628
Jonathan Burian, Region 5
Burian.Jonathan@epa.gov
Laura Webb, Region 7
Webb.Laura@epa.gov
Bill Schroder, Region 8
Ryan Monahan, Region 8
Schroeder.william@eoa.gov
303-462-9472
Monahan.rvan@eoa.gov
303-462-9477
Peter Husby, Region 9
Husby.peter@epa.gov
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Title
Name
Contact Information
510-412-2311
Bob Hopeman, Region 9
Hopeman. Bob(ฎ eoa.gov
510-415-2388
Peter Leinenbach, Region 10
leinenbach.peter(ฎ eoa.gov
*Regional NLA coordinator and NES field crew lead
Project and Data Quality Objectives
NES dataset provides a unique opportunity to assess change in lake condition in the intervening 50 years
(1972, 2007, 2012, 2017, and 2022). In addition to the analysis of the filed collected data, analyses will
include information on the types of management practices implemented at and near the lakes
(wastewater treatment, etc.), population changes and other stressors to evaluate water quality changes
in lakes with restoration and protection activities.
The NES 2022 resampling study is designed to address the following objectives:
1. How as the trophic condition of the NES lakes changed in the last 50 years (1972, 2007, 2012,
2017, and 2022). Are conditions better or worse?
2. Have stressors within the watersheds changed since the 1970s?
3. What stressors contribute to the current trophic condition of the NES lakes?
NES Indicators
The original NES study included the collection of lake profile data, water chemistry, chlorophyll -a, and
secchi. The NES 2022 resurvey will continue to focus on measurements of trophic condition and will
include the NLA indicators identified in Table D-2.
Table D-2 NLA 2022 NES site water quality indicators.
Indicator Type
Indicator
NES
1970s
NLA/NES
2007
NLA/NES 2022
Trophic and
Chemical
Vertical profile measurements (DO,
Temperature, pH)
X
X
X
Indicators
Secchi disk transparency
X
X
X
Water chemistry (NH4, N03), major
X
X
X
anions and cations, alkalinity (ANC),
DOC, TSS, silica, conductivity,
nutrients (total and dissolved TN
and TP)
Chlorophyll-o
X
X
X
Human Health
CyanoHAB visual observations at
index site and boat launch
X
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Indicator Type
Indicator
NES
1970s
NLA/NES
2007
NLA/NES 2022
Algal toxins (microcystins and
X
cylindrospermopsin)
Phytoplankton (cyanobacteria cell
X
X
count)
eDNA
Water chemistry
Other Indicators
filter only
Lake area, basin morphometry, and
characteristics of watershed
X
X
X
Study Design
In NLA 2007, 210 NES lakes were included in the survey design and resampled. This subpopulation was
used to report on the changes in trophic condition in the 2007 report. The NES 2022 design is a random
design of the NES 2007 lakes stratified by EPA Region. Due to resource limitation, all 210 lakes cannot be
sampled, therefore we identified the target number of lakes in each Region (Table D-3). In Regions with
12 or less lakes, all NES lakes will be targeted for sampling. In Regions with greater than 12 sites, 60% of
the lakes will be targeted for sampling. Note that 3 lakes will likely be sampled as part of the NLA
probabilistic sampling efforts. The selection of target lakes must happen in the order of the site ID;
however, sampling may happen in any order.
Table D-3 Target number of hand-picked lakes for sampling by EPA Region.
Row Labels
NES Lakes
per
Region
Target # of
Sites
Lakes in the
NLA 2022
Design
Total #of
Target Lakes by
Region
Regional
sites
Contractor
Sites
Region_l
5
5-6
1 (likely)
6
5
0
Region_2
6
6
6
6
0
Region_3
9
9
9
10**
0
Region_4
35
24*
1
25
24*
0
Region_5
48
29
29
28**
0
Region_6
32
19
19
5
14
Region_7
10
10
10
10
0
Region_8
38
23
23
9
14
Region_9
13
13
13
8
5
Region_10
12
11
1
12
6
5
Total
206
146
3
152
82
38
* Region 4 will sample the first 3 oversample lakes given there proximity to the target lakes, increasing the total
number of lakes sampled by R4 to 24.
**One OH lake in Region 5 will be sampled by R3.
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Field Methods and Procedures
Field Crew Training
The NES field crew leads and crew members are encouraged to participate in all portions of the NLA
2022 training. In the event crew members are not able to complete the full training program, below is
the required list of training material for NES sampling teams:
1. Review the NLA QAPP, SEG and FOM (see Daily Field Activities below for FOM details)
2. Review the following training videos and take the associated quizzes:
a. Block 1 - Background (all 4 videos)
b. Block 2 - Launch sites activities and index sampling (videos 2-1 through 2-5; no
zooplankton)
c. Block 5 - Final lake activities (all 3 videos)
3. Participate in a Q&A session
4. Attend a regional in-person training (requirement for field crew leads only)
a>
The NLA also includes Endangered Species Act (ESA) training for federal field crews. Although the NES ^
>
"O
-M
LD
C
o
sampling does not include the NLA biological indicators that are most likely to effect ESA listed species
(i.e., benthic macroinvertebrates and fishing), all NES crew leads are to participate in the training in the
event listed species are encountered while in the field.
i. NES sampling will not include the collection of eDNA, atrazine, and zooplankton
samples
Field Forms and Sample Shipment
samples that are not part of NES field work. For the samples that are not collected at NES lakes (i.e.,
atrazine, all littoral samples, eDNA, zooplankton), the field crew is to identify that no sample was
collected in the assoiceated forms. The Physical Habitat form will not be submitted.
Daily Field Activities
The NES sampling will include the following field activities: o
1. All base site activities (see Section 4.0). =
2. The following index site activities (see Section 5.0): "js
a. CyanoHABS Visual Assessment ~
CD
b. Temperature, DO and pH profile z
c. Secchi Dish Transparency J:
d. Water sample collection for chlorophyll-a, phytoplankton, algal toxins and water 'o
chemistry ,c
"5.
E
tc
Q.
0)
All final lake activities ^
In
a. Filtering only includes the chlorophyll-a sample (see Sections 8.2.3 and 8.2.4) ฃ
"O
Q)
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Table D-4 identifies the NLA field forms that will be used for the NES sampling. Many forms will include 1
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115
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Table D-4 NES field forms.
Form
Fill out
Submit?
Verification
Entire form
Yes
Calibration
Entire form
Yes
Index Samples
Entire form;
mark FDNA, TRIA, ZOCN, and ZOFN as not collected
Yes
Profile
Entire form
Yes
Littoral Samples
Bottom of form only;
mark BENT, LDNA, and ENTE as not collected
Yes
Physical Habitat
None
No
Assessment
Entire form
Yes
Whole Fish Sample
Top of form only;
mark 'No'
Yes
Federal ESA
Fill in a pertinent section if an ESA-listed species is encountered
If data are entered
Tracking
Tl, T2, and T3 sections when shipping samples from those
groups
Yes
Sample shipments will include:
T-l (daily water chemistry samples);
T-2 (frozen batched samples - algal toxins only); and
T-3 (non-chilled batched samples - phytoplankton only).
See APPENDIX B: SHIPPING GUIDELINES for additional details.
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