EPA Publication LPA 841 -R-24-006
National Lakes Assessment 2022:
Technical Support Document
U.S. Environmental Protection Agency
Office of Wetlands, Oceans and Watersheds
Office of Research and Development
Washington, DC 20460
August 2024
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NLA 2022 Technical Support Document - August 2024
Suggested citation for this document is: U.S. Environmental Protection Agency. 2024. National Lakes
Assessment 2022: Technical Support Document. EPA 841 -R-24-006. U.S. Environmental Protection
Agency, Office of Water and Office of Research and Development.
This report is available here on the NLA Website.
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Table of Contents
Chapter 1: Project Overview 12
1.1 Overview 12
1.2 Objectives of the National Lakes Assessment 12
1.3 Considerations for the NLA 2022 TSD and public report 13
Chapter 2: Survey Design and Population Estimates 14
2.1 Description of sample design 14
2.1.1 Stratification 14
2.1.2 Unequal probability categories 14
2.1.3 Fish Tissue Study 15
2.1.4 Panels 15
2.1.5 Expected sample size 16
2.2 Sampling frame summary 19
2.3 Survey design implementation and analysis 21
2.4 Estimated number of the NLA lakes and implications for reporting 22
2.5 Literature cited 23
Chapter 3: Defining Reference Sites and Condition 25
3.1 Background information 25
3.2 Pre-sampling screening (hand-picked sites only) 26
3.3 Post-sampling screening for biological reference condition 27
3.4 Literature cited 30
Chapter 4: Benthic Macroinvertebrates 32
4.1 Background information 32
4.2 Data preparation 32
4.2.1 Standardizing counts 32
4.2.2 Autecological characteristics 32
4.2.3 Tolerance values 33
4.2.4 Functional feeding group and habitat preferences 33
4.2.5 Taxonomic resolution 33
4.3 Multimetric index development 34
4.3.1 Dataset 34
4.3.2 Low macroinvertebrate numbers 34
4.3.3 Ecoregion classification 34
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4.3.4 Metric screening 34
4.3.5 All Subsets MMI selection 35
4.3.6 Setting MMI benchmarks 39
4.4 Literature cited 39
Chapter 5: Physical Habitat 40
5.1 Background information 40
5.2 Data preparation 41
5.3 Methods 42
5.3.1 NLA sites used for expected condition modeling and precision estimates 42
5.3.2 Field sampling design and methods 42
5.3.3 Classifications 43
5.3.4 Calculation of lake physical habitat metrics 44
5.3.5 Calculation of summary physical habitat condition indices 51
5.3.6 Deriving expected index values under least disturbed conditions 55
5.3.7 Condition criteria for nearshore lake physical habitat 57
5.4 Least disturbed reference distributions and regressions (from sections 5.3.6 and 5.3.7) 59
5.4.1 Disturbance within least disturbed reference sites 59
5.4.2 Null model results for RVegQ, LitCvrQ, and LitRipCvQ: 60
5.4.3 O/E model results for RVegQ, LitCvrQ, and LitRipCvQ: 60
5.4.4 Null model results for lake drawdown and level fluctuations: 61
5.5 Precision of physical habitat indicators 62
5.6 Physical habitat index responses to anthropogenic disturbance 63
5.7 Discussion 64
5.8 Literature cited 65
Chapter 6: Water Chemistry 84
6.1 Background information 84
6.2 Chemical condition benchmarks 84
6.2.1 Acidity 84
6.2.2 Dissolved Oxygen 84
6.2.3 Trophic State 85
6.2.4Total nitrogen, total phosphorus, chlorophyll a, and turbidity 85
6.2.5 Atrazine 88
6.2.6 Within-year variability 89
6.3 Literature cited 90
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Chapter 7: Zooplankton 91
7.1 Background information 91
7.2 Methods 92
7.2.1 Field methods 92
7.2.1 Laboratory methods 94
7.3 Data preparation 95
7.3.1 Data quality assurance 95
7.3.2 Master taxa list 95
7.3.3 Aggregations and rarefaction of count data 96
7.4 Zooplankton MMI development 96
7.4.1 Regionalization 96
7.4.2 Least and most disturbed sites 97
7.4.3 Least disturbed sites: calibration versus validation 98
7.4.4 Candidate metrics 98
7.4.5 Final metric selection 99
7.4.6 Metric scoring 101
7.5 Zooplankton MMI metric composition and performance 101
7.5.1 Coastal Plains MMI 101
7.5.2 Eastern Highlands MMI 103
7.5.3 Plains MMI 105
7.5.4 Upper Midwest MMI 107
7.5.5 Western Mountains MMI 107
7.6Zooplankton MMI performance Ill
7.6.1 Calibration versus validation sites Ill
7.6.2 Precision of MMIs based on least disturbed sites Ill
7.6.3 Responsiveness, redundancy, and repeatability of zooplankton MMIs Ill
7.6.4 Responsiveness to a generalized stressor gradient 114
7.6.5 Effect of natural drivers and tow length on MMI scores 114
7.7 Thresholds for assigning ecological condition 119
7.7.1 NLA 2012 119
7.7.2 NLA 2017 122
7.7.3 NLA 2022 123
7.8 Discussion 123
7.9 Literature cited 126
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Chapter 8: Human Health Water Quality Indicators 130
8.1 Enterococci indicator 130
8.1.1 Field collection 130
8.1.2 Lab methods 130
8.1.3 Analysis and application of benchmarks 131
8.2 Cyanobacteria toxins (Cyanotoxins) 131
8.2.1 Field methods 132
8.2.2 Analysis and application of benchmarks 132
8.3 Literature cited 134
Chapter 9: Human Health Fish Tissue 135
9.1 Field fish collection 135
9.2 Mercury analysis and fish tissue screening levels to protect human health 136
9.3 PCB analysis and fish tissue screening levels to protect human health 137
9.4 PFAS analysis and fish tissue screening levels to protect human health 137
9.5 Calculation of fish tissue screening levels for human health protection 139
9.6 Literature cited 141
Chapter 10: From Analysis to Results 142
10.1 Background information 142
10.2 Population estimates 142
10.2.1 Subpopulations 142
10.3 Lake extent estimates 145
10.4 Stressor extent, relative risk, and attributable risk 145
10.4.1 Stressor extent 146
10.4.2 Relative risk and attributable risk 146
10.4.3 Relative risk 147
10.4.4 Attributable risk 147
10.4.5 Considerations when calculating and interpreting relative risk and attributable risk 148
10.5 Change analysis 149
10.5.1 Background information 149
10.5.2 Data preparation 150
10.5.3 Methods 150
10.6 Literature cited 151
Chapter 11: Quality Assurance Summary 152
Appendix A: Lake Physical Habitat Expected Condition Models 156
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Appendix B: Survey Design and Estimated Extent Summary for NLA 2007, 2012, 2017 and 2022 196
Appendix C: NLA 2022 Indicator Benchmark Summary 199
Appendix D: Zooplankton 206
11.1 List of candidate metrics for zooplankton 206
11.2 Non-target taxa in zooplankton samples that are excluded from enumeration 227
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List of Figures
Figure 2.1. The number of lake objects in the NLA 2022 sampling frame, evaluated lakes, sampled lakes
and lakes in the NLA target population 23
Figure 3.1. Nine aggregate ecoregions used for reference site classification 27
Figure 4.1. Box and whisker plots showing discrimination between least disturbed reference (L) and
most disturbed (M) sites by biological ecoregion in the NLA 2007-2012 data used to develop the
MMI. Boxes show the interquartile range and the whiskers show the 5th and 95th percentiles.
Outliers are not presented 38
Figure 5.1. Field sampling design with 10 near-shore stations at which data were collected to
characterize near shore lake riparian and littoral physical habitat in the 2007 and 2012 National
Lakes Assessment (NLA) surveys. The 10 stations were systematically spaced around the shore
of the lake from random starting point. Insert shows riparian plot, shoreline band, littoral plot,
and (for NLA 2012 and later) drawdown zone plot located at each station 78
Figure 5.2. Near-shore anthropogenic disturbance (RDis_IX) in NLA0712 regions, ordered by their
median Reference site RDis 79
Figure 5.3. Near-shore anthropogenic disturbance in NLA0712 least disturbed reference sites (median
RDisJX), ordered by aggregated region according to the same median level of near-shore
disturbance 80
Figure 5.4. LogSD's for Null-Model and regression-based O/E model for Near-shore RVegQ, LitCvrQ, and
LitRipCvrQ in the set of least disturbed lakes and reservoirs (Table 5-1) sampled in the
combined NLA 2007 and 2012 surveys 81
Figure 5.5. Contrasts in key NLA physical habitat index values among least disturbed reference (L),
intermediate (I), and most disturbed (M) lakes in the contiguous 48 states of the U.S. based on
combined NLA 2007 and 2012 data. Unweighted sample statistics are shown; box midline and
lower and upper ends show median and 25th and 75th percentile values, respectively; whiskers
show maximum and minimum observations within 1.5 times the interquartile range above/
below box ends; circles show outliers. See Table 5-9 for t and p values for the differences
between means for least disturbed reference (L) and most disturbed (M) sites 82
Figure 5.6. Contrasts in key NLA physical habitat index values among least disturbed reference (L),
intermediate (I), and most disturbed (M) lakes in the contiguous 48 states of the U.S. shown
separately for the NLA 2007 and 2012 surveys 83
Figure 6.1. Box and whisker plot of Total Phosphorus in GIS screened, outlier removed, 2007-2017
nutrient reference sites by ecoregion 87
Figure 6.2. Box and whisker plot of Total Nitrogen in GIS screened, outlier removed, 2007-2017
nutrient reference sites by ecoregion 87
Figure 7.1 Five aggregated bio-regions used to develop zooplankton MMIs for the 2012 National Lake
Assessment (CPL=Coastal Plains; EHIGH=Eastern Highlands, PLAINS= Plains, UMW=Upper
Midwest, and WMTNS=Western Mountains). Solid dots indicate least disturbed sites used for
developing the zooplankton MMI. White circles indicate least disturbed sites that we excluded
because of atypical samples (too few taxa or number of individuals collected) 97
Figure 7.2. Distribution of six component metrics of the zooplankton MMI for the Coastal Plains bio-
region in least disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th
percentiles 102
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Figure 7.3 Distribution of six component metrics of the zooplankton MMI for the Eastern Highlands bio-
region in least disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th
percentiles 104
Figure 7.4. Distribution of six component metrics of the zooplankton MMI for the Plains bio-region in
least disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th percentiles.
106
Figure 7.5. Distribution of six component metrics of the zooplankton MMI for the Upper Midwest bio-
region in least disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th
percentiles 108
Figure 7.6. Distribution of six component metrics of the zooplankton MMI for the Western Mountains
bio-region in least (L) disturbed versus most disturbed (M) sites. Dots indicate the 5th and 95th
percentiles 110
Figure 7.7. Distribution of zooplankton MMI scores in-calibration vs. validation sites for five bio-
regions. Sample sizes are in parentheses. Dots indicate the 5th and 95th percentiles 112
Figure 7.8 Distribution of zooplankton MMI scores in least-disturbed (L) vs. most disturbed (M) sites for
five bio-regions. Sample sizes are in parentheses. Dots indicate the 5th and 95th percentiles.
113
Figure 7.9. Linear regression of NLA 2012 Zooplankton MMI scores vs. first axis score from principal
components analysis (PCA) based on chemical, habitat, and visual assessment stressor variables
used to screen least- and most disturbed sites 115
Figure 7.10. NLA 2012 Zooplankton MMI scores of human-made (shaded boxes) versus natural lakes
(unshaded boxes) for least disturbed sites in five bio-regions. See Figure 7.1 for bio-region
codes. Sample sizes for each type are in parentheses. Dots indicate 5th and 95th percentiles.. 115
Figure 7.11. Zooplankton MMI scores versus lake size class within least disturbed lakes of the NLA
2012. Sample sizes are in parentheses. Dashed lines are mean values. Dots indicate the 5th and
95th percentiles 116
Figure 7.12. Zooplankton MMI scores versus site depth for least disturbed sites 120
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List of Tables
Table 2-1. National Lakes Assessment 2022 Initial Design. The number of lakes to be sampled by state
and the final design by aggregated ecoregion 16
Table 2-2. Actual number of sites sampled for NLA 2022 by design categories, including state
intensification sites that were used in the national condition estimate analyses. Two sampled
sites were determined to be non-target and removed from the national analyses 18
Table 2-3 Number of waterbody objects in NHDPIusHR by type and sampling frame inclusion 20
Table 2-4 Number of lake objects in the sampling frame by aggregated ecoregion and lake area
category 21
Table 3-1. Least disturbed reference screening filter thresholds for NLA 2017 28
Table 3-2. Most disturbed site screening thresholds for NLA 2017 29
Table 3-3. Dichotomous key for defining NLA lakes likely impacted by anthropogenic drawdown 30
Table 4-1. Final NLA biological ecoregion benthic MMI metrics and their floor/ceiling values for MMI
scoring 36
Table 4-2. Benthic MMI statistics for the NLA 2007-2012 data used to develop the MMI 38
Table 4-3. Macroinvertebrate MMI benchmarks using 2007-2017 reference site data 39
Table 5-1. NLA reference sites from combined 2007 & 2012 surveys 70
Table 5-2. Assignment of riparian vegetation cover complexity, littoral cover complexity, and littoral-
riparian habitat complexity index variants by aggregated ecoregion 70
Table 5-3. Summary of regression models used in estimating lake-specific expected values of Lake
Physical Habitat variables RVegQx, LitCvrQx and LitRipCvrQx under least disturbed conditions.71
Table 5-4. Null Model Geometric Means (gMean), geometric Standard Deviations (gSD), 5th percentiles,
and 25th percentiles of habitat index values in least disturbed reference lakes in the aggregated
ecoregions of the NLA 72
Table 5-5. O/E Physical Habitat Model means (LogMean, gMean), standard deviations (LogSD, gSD),
and percentiles of the distribution of habitat index O/E values for least disturbed reference
lakes in the aggregated ecoregions of the NLA 73
Table 5-6. Empirical 75th and 95th percentiles of the distribution of vertical and horizontal drawdown.74
Table 5-7. Precision of the key NLA Physical Habitat indices used as the primary physical habitat
condition measures in the NLA 75
Table 5-8. Association of NLA-2012 Physical Habitat Indices with high and low anthropogenic
disturbance stress classes (RT_NLA12 = R and T), defined as least disturbed and most disturbed
within NLA regions 76
Table 5-9. Association of NLA 2007 and 2012 Physical Habitat Indices with high and low anthropogenic
disturbance stress classes (RT_NLA12 = L and M), defined as least disturbed and most disturbed
within NLA regions 77
Table 6-1. Trophic State Classification used in NLA 85
Table 6-2 Overall S:N and pooled standard deviation (SD) for NLA 2007 and 2012 surface water
chemistry within three concentration range classes. N = 192 89
Table 6-3. Atrazine detection (a) and risk condition (b) contingency tables. N = 293 90
Table 7-1. Hypothesized responses of zooplankton assemblages to disturbance 93
Table 7-2. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE COASTAL PLAINS BIO-REGION.
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Table 7-3. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE EASTERN HIGHLAND BIO-
REGION 104
Table 7-4. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE PLAINS BIO-REGION 106
Table 7-5. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE UPPER MIDWEST BIO-REGION.
108
Table 7-6. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE WESTERN MOUNTAINS BIO-
REGION 110
Table 7-7. RESULTS OF INDEPENDENT ASSESSMENT AND PRECISION TESTS OF NLA 2012
ZOOPLANKTON MMIs BASED ON LEAST DISTURBED SITES 112
Table 7-8. RESULTS OF RESPONSIVENESS, REDUNDANCY, AND REPEATABILITY TESTS FOR NLA 2012
ZOOPLANKTON MMIs 113
Table 7-9. Component metrics of the zooplankton multimetric indices (MMIs) used for NLA 2022. ...117
Table 7-10. LINEAR REGRESSION STATISTICS OF ZOOPLANKTON MMI SCORES VERSUS PCA-BASED
DISTURBANCE SCORE FOR EACH BIO-REGION 121
Table 7-11. ECOLOGICAL CONDITION BENCHMARKS FOR ZOOPLANKTON MMI SCORES (NLA 2012
ONLY) BASED ON THE DISTRIBUTION OF LEAST DISTURBED SITES IN FIVE BIO-REGIONS 121
Table 7-12 Ecological condition benchmarks for NLA 2017 zooplankton MMI scores based on the
distribution of least disturbed sites in five aggregated ecoregions (bio-regions) 122
Table 8-1 Enterococci condition contingency table; N = 96 131
Table 8-2. Microcystin detection (a) and risk condition (b) contingency tables; N = 293 133
Table 8-3. Cylindrospermopsin detection (a) and risk condition (b) contingency tables; N = 193 133
Table 9-1. Primary and secondary NLA target species for human health fish collection 136
Table 9-2. NLA 2022 fish tissue fillet composite sample summary data 138
Table 9-3. NLA 2022 fish fillet tissue sampled population exceedances for mercury and total
polychlorinated biphenyls (PCBs) 140
Table 10-1. Extent estimates for response and stressor categories 147
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Chapter 1: Project Overview
1.1 Overview
This document, the National Lakes Assessment 2022: Technical Support Document,
accompanies the National Lakes Assessment: The Fourth Collaborative Survey of Lakes in the
United States and related on-line materials. The National Lakes Assessment (NLA) is a
collaboration among the U.S. Environmental Protection Agency (EPA), states, tribes, and other
partners. It is part of the National Aquatic Resource Surveys (NARS) program design to conduct
national scale assessments of aquatic resources. The NLA 2022 provides condition assessment
results at national and regional scales of the ecological and recreational condition of lakes. This
assessment was accomplished by collecting and analyzing data from across the conterminous
United States.
The National Lakes Assessment: The Fourth Collaborative Survey of Lakes in the United States
(the public report) is not a technical document, but rather a report geared toward a broad,
public audience. It provides national-scale assessments and compares the condition of lakes to
those from the earlier NLAs (2007, 2012, 2017) conducted by EPA and its partners. You can find
results for regional scales and comparisons between natural lakes and reservoirs using the NLA
2022 interactive dashboard. This document serves as a technical reference to support findings
presented in the public report and on-line.
1.2 Objectives of the National Lakes Assessment
The objective of the NLA is to characterize aspects of the biological, chemical, physical, and
recreational condition of the nation's lakes throughout the conterminous United States. It
employs a statistically valid probability design stratified to allow estimates of the condition of
lakes on a national and regional scale.
The NLA is designed to answer the following questions about lakes across the United States.
1. What is the current biological, chemical, physical, and recreational condition of lakes?
a. What is the extent of degradation among lakes?
b. Is degradation widespread (e.g., national) or localized (e.g., regional)?
2. Is the proportion of lakes in the poor condition getting better, worse, or staying the same
over time?
3. Which environmental stressors are most strongly associated with degraded biological
condition in lakes?
A variety of chemical, physical, and biological data were collected and developed into indicators
to address the NLA questions. For each of these indicators, this Technical Report focuses on the
conceptual basis, methods, and procedures used for the NLA. The information described in this
Technical Report was developed through the efforts and cooperation of NLA scientists from
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EPA, technical experts, and participating cooperators from states, tribes, and academia. While
this Technical Report serves as a comprehensive summary of the NLA procedures, it is not
intended to present an in-depth report of the design, site evaluation process, field sampling,
NLA results, or additional data analysis results. Please see the following documents for
additional details on these aspects of the project.
• National Lakes Assessment 2022: Quality Assurance Project Plan (EPA 841-B-21-
009)(hereafter referred to as the NLA 2022 QAPP)
• National Lakes Assessment 2022: Site Evaluation Guidelines (EPA 841-B-21-008)
(hereafter referred to as the NLA 2022 SEG)
• National Lakes Assessment 2022: Field Operations Manual (EPA 841-B-21-011)
(hereafter referred to as the NLA 2022 FOM)
• National Lakes Assessment 2022: Laboratory Operations Manual (EPA 841-B-21-010)
(hereafter referred to as the NLA 2022 LOM)
1.3 Considerations for the NLA 2022 TSD and public report
The EPA is working to stabilize benchmarks and data analyses across the NARS program to
facilitate change and trend analyses. In NLA 2022, most aspects of the survey remained the
same including the field methods, laboratory analyses, target population, benchmark selection
process and data analyses. Changes since the NLA 2017 that are discussed in this document
include:
• Updated sampling frame that uses NHDPIus High Resolution for all new lakes (see
Chapter 2);
• The lake drawdown calculations and results are presented as small, medium and large
conditions categories, in 2017 the categories included not large and large drawdown
(see Chapter 5);
• The addition of enterococci and cylindrospermopsin (see Chapter 8); and
• The addition of Human Health Fish Tissue Indicator (see Chapter 9).
For purposes of identifying change and trends, prior survey results were recalculated based on
updated 2022 benchmarks (see Appendix C) as needed. Given the above modifications, direct
comparisons should not be made between the NLA 2022 results and those reported in earlier
surveys as this will produce erroneous information.
Finally, the NLA 2022 public report and this document use the good/fair/poor terminology for
condition class estimates consistent with the public report. Least, moderate, and most
disturbed condition classes are also used in this document to describe anthropogenic
disturbance pressure categories for index and model development (see Chapters 4, 5, and 7).
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Chapter 2: Survey Design and Population Estimates
The NLA was designed to assess the condition of the population of lakes, reservoirs, and ponds
in the conterminous United States. The NLA design allows characterization of lakes at national
and regional scales using chemical, physical and biological indicators. It is not intended to
represent the condition of individual lakes. The statistical design also accounts for the
distribution of lakes across the country - some areas have fewer lakes than others - so that
even in areas of the country where there are few sample sites regional and national results still
apply to the broader target population.
This chapter provides details on the NLA survey design, sampling frame, analyses and estimated
extent of the NLA lake population. Modifications to the survey design in 2022 are noted
throughout the chapter and are summarized in Appendix B: Survey Design Summary and
Population Estimates for NLA 2007, 2012, 2017, and 2022.
2.1 Description of sample design
The target population for the NLA includes all lakes, reservoirs, and ponds within the 48
contiguous United States greater than 1 hectare (ha) in surface area that are permanent
waterbodies, at least 1 meter deep, and have a minimum 0.1 ha of open water. In addition,
lakes are required to have a minimum residence time of one week. The word "lake" in the
remainder of this document includes lakes, reservoirs and ponds. The Great Lakes, Great Salt
Lake and lakes that are tidally influenced are excluded; as are those used for aquaculture,
disposal-tailings, sewage treatment, evaporation, or other unspecified disposal use.
NLA 2022 uses a spatially balanced survey design where lakes are viewed as a finite population
(i.e., each lake is viewed as a point identified by the centroid of the lake polygon). To select
sites for the NLA, EPA statisticians used a Generalized Random Tessellation Stratified (GRTS)
(Stevens and Olsen 2004; Olsen et al. 2012) survey design for a finite resource with
stratification and unequal probability of selection.
2.1.1 Stratification
The design is stratified by state. Within each state, lakes are selected using unequal probability
categories based on lake area.
2.1.2 Unequal probability categories
Unequal probability categories used for the NLA 2017 subsample were defined based on lake
area: 1 to 4 ha, 4 to 10 ha, 10 to 20 ha, 20 to 50 ha and greater than 50 ha. For new NLA 2022
lakes, the unequal probability categories included 1 to 4 ha, 4 to 10 ha, 10 to 50 ha and greater
than 50 ha. The collapsing to four lake area categories reflects that no differences in percent of
non-target lakes nor in landowner access were found. Given that weight adjustment on all
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evaluated sites is likely to use lake area categories, having fewer categories will result in more
stable weight adjustments since they will be based on more evaluated lakes within a category.
2.1.3 Fish Tissue Study
A subset of the lakes selected using the above survey design will have fish sampled for the
analysis offish tissue contaminants. The subsample is approximately 2/3 of the base lakes
selected for the main NLA 2022 survey. Approximately 50% of the lakes will be from the
subsample of NLA 2017 lakes and 50% from new lakes selected for 2022. These lakes will be
assigned to panels that will identify them.
2.1.4 Panels
The survey design incorporates lakes sampled in in prior NLAs as well as selecting new lakes.
This improves the ability of the survey design to estimate change in condition in NLA 2022 from
the condition in prior surveys. In addition, the survey design includes 96 lakes that are sampled
twice in NLA 2022, providing information on measurement variability. These requirements
result in five base and two oversample panels:
• NLA22_17RVT2FT - Panel of lakes originally sampled in NLA 2017. These lakes will be
sampled twice in NLA 2022 for all indicators except for fish tissue which will be sampled
for only one of the two visits.
• NLA22_17BaseFT - Panel of lakes originally sampled in NLA 2017 and will be sampled
once in NLA 2022 for all indicators as well as fish tissue.
• NLA22_17Base - Panel of lakes originally sampled in NLA 2017 and will be sampled once
in NLA 2022 for all indicators except fish tissue.
• NLA22_22BaseFT - Panel of new lakes to be sampled once in NLA 2022 for all indicators
including fish tissue.
• NLA22_22Base - Panel of new lakes to be sampled once in NLA 2022 for all indicators
except fish tissue.
• NLA22_170ver - Over sample lakes to be used as replacements for NLA22_17RVT2FT or
NLA22_17BaseFT or NLA22_17Base lakes when they cannot be sampled for any reason.
If the lake being replaced was scheduled to be sampled for fish tissue, then the
replacement lake will be sampled for fish tissue.
• NLA22_220ver - Over sample lakes to be used as replacements for NLA22_22BaseFT or
NLA22_22Base lakes when they cannot be sampled for any reason. If the lake being
replaced was scheduled to be sampled for fish tissue, then the replacement lake will be
sampled for fish tissue.
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2.1.5 Expected sample size
For NLA 2022, 904 lakes will be sampled with 96 of the lakes sampled twice for a total of 1000
lake visits. Consequently, 904 unique sites will be sampled with 808 sampled only once and 96
sites being sampled twice during 2022 resulting in 1000 (808 + 2*96) total site visits. Reporting
will be nationally as well as for nine aggregated ecoregions (CPL, NAP, SAP, UMW, NPL, SPL,
TPL, WMT and XER). Approximately, 100 lakes will be sampled in each aggregated ecoregion.
For each aggregated ecoregion, the number of lakes assigned to each state within the
ecoregion will be proportional to the number of lakes in the sampling frame within the state.
The total lakes for a state will be the sum across all ecoregions in the state. In addition, the
minimum number of lakes for a state will be 8 and the maximum will be 50. With these
constraints and with proportional allocation, two states (TX and MN) are allocated more than
50 lakes and 13 states (AZ, CT, DE, IA, MD, NH, NJ, NM, NV, Rl, TN, VT, WV) have 8 or fewer. For
these states, lakes in the sampling frame are allocated by ecoregion within each state to get
minimum of 8 and maximum of 50. Then the remaining states are re-allocated lakes by
ecoregion to satisfy the total sample size. The final allocation by state and aggregated
ecoregion is given in Table 2-1. Approximately 50% of the lakes will be lakes sampled in NLA
2017 that were sampled as new lakes in 2017.
The survey design does not select lakes based on aggregated ecoregions; only the total number
of lakes for a state is specified in the survey design. For new lakes, approximately an equal
number of lakes by the four lake area categories are selected with unequal probability within
each state. For lakes sampled as new lakes in 2017, the lakes selected for 2022 are the first
lakes evaluated in 2017 to meet the sample size requirement for 2017 lakes to be resampled in
2022. Note that these are the expected number of lakes and not the final number of lakes
selected by the survey design (see section "Final Survey Design Summary").
Table 2-1. National Lakes Assessment 2022 Initial Design. The number of lakes to be sampled by state and the final
design by aggregated ecoregion.
State
CPL
NAP
NPL
SAP
SPL
TPL
UMW
WMT
XER
Total
AL
11
0
0
3
0
0
0
0
0
14
AR
7
0
0
3
0
0
0
0
0
10
AZ
0
0
0
0
0
0
0
4
4
8
CA
0
0
0
0
0
0
0
25
23
48
CO
0
0
0
0
9
0
0
8
2
19
a
0
8
0
0
0
0
0
0
0
8
DE
7
0
0
1
0
0
0
0
0
8
FL
11
0
0
0
0
0
0
0
0
11
GA
22
0
0
11
0
0
0
0
0
33
IA
0
0
0
0
0
7
2
0
0
9
ID
0
0
0
0
0
0
0
10
6
16
IL
0
0
0
1
0
15
1
0
0
17
IN
0
0
0
2
0
16
5
0
0
23
KS
0
0
0
0
5
14
0
0
0
19
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NLA 2022 Technical Support Document - August 2024
State
CPL
NAP
NPL
SAP
SPL
TPL
UMW
WMT
XER
Total
KY
0
0
0
8
0
1
0
0
0
9
LA
13
0
0
0
0
0
0
0
0
13
MA
1
7
0
0
0
0
0
0
0
8
MD
6
0
0
2
0
0
0
0
0
8
ME
0
15
0
0
0
0
0
0
0
15
Ml
0
0
0
0
0
1
28
0
0
29
MN
0
0
0
0
0
3
48
0
0
51
MO
2
0
0
5
0
7
0
0
0
14
MS
11
0
0
0
0
0
0
0
0
11
MT
0
0
28
0
0
0
0
17
0
45
NC
5
0
0
7
0
0
0
0
0
12
ND
0
0
23
0
0
15
0
0
0
38
NE
0
0
0
0
24
5
0
0
0
29
NH
0
8
0
0
0
0
0
0
0
8
NJ
4
0
0
4
0
0
0
0
0
8
NM
0
0
0
0
2
0
0
2
4
8
NV
0
0
0
0
0
0
0
0
8
8
NY
1
28
0
1
0
0
0
0
0
30
OH
0
4
0
3
0
6
0
0
0
13
OK
2
0
0
5
23
5
0
0
0
35
OR
0
0
0
0
0
0
0
15
6
21
PA
0
7
0
6
0
0
0
0
0
13
Rl
1
8
0
0
0
0
0
0
0
9
SC
8
0
0
0
0
0
0
0
0
8
SD
0
0
18
0
0
21
0
1
0
40
TN
4
0
0
4
0
0
0
0
0
8
TX
26
0
0
0
23
0
0
0
1
50
UT
0
0
0
0
0
0
0
6
8
14
VA
4
0
0
7
0
0
0
0
0
11
VT
0
8
0
0
0
0
0
0
0
8
WA
0
0
0
0
0
0
0
20
7
27
Wl
0
0
0
0
0
4
22
0
0
26
WV
0
0
0
8
0
0
0
0
0
8
WY
0
0
5
0
2
0
0
11
8
26
Sum
146
93
74
81
88
120
106
119
77
904
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NLA 2022 Technical Support Document - August 2024
Table 2-2. Actual number of sites sampled for NLA 2022 by design categories, including state intensification sites
that were used in the national condition estimate analyses. Two sampled sites were determined to be non-target
and removed from the national analyses.
State
CPL
NAP
NPL
SAP
SPL
TPL
UMW
WMT
XER
Total
AL
10
4
14
AR
8
2
10
AZ
4
4
8
CA
20
26
46
CO
6
13
19
a
8
8
DE
7
1
8
FL
11
11
GA
15
18
33
IA
8
8
ID
10
6
16
IL
1
16
17
IN
7
33
10
50
KS
4
16
20
KY
1
7
1
9
LA
13
13
MA
3
6
9
MD
2
6
8
ME
15
15
Ml
4
46
50
MN
10
40
50
MO
4
10
14
MS
11
11
MT
23
22
45
NC
3
9
12
ND
28
12
40
NE
1
17
11
29
NH
8
8
NJ
4
1
3
8
NM
2
3
3
8
NV
1
7
8
NY
1
29
30
OH
7
2
4
13
OK
1
4
25
5
35
OR
18
3
21
PA
9
4
13
Rl
1
7
8
SC
3
5
8
SD
12
33
1
46
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NLA 2022 Technical Support Document - August 2024
State
CPL
NAP
NPL
SAP
SPL
TPL
UMW
WMT
XER
Total
TN
4
4
8
TX
21
27
2
50
UT
13
1
14
VA
5
6
11
VT
8
8
WA
16
11
27
Wl
4
46
50
WV
8
8
WY
3
1
17
5
26
Sum
124
98
67
95
82
167
142
138
68
981
2.2 Sampling frame summary
The sampling frame was derived from the National Hydrography Dataset Plus High Resolution
(NHDPIus HR) data layer. The total number of waterbody polygons in NHDPIus HR is 6,512,454,
which includes several non-target waterbody types (e.g., swamp/marsh, estuary, etc).
Attributes were created to identify the polygons that are lakes to included in the sampling
frame and those to exclude from the sampling frame. First, polygons that were less than or
equal to 1 hectare were excluded. Next polygons were included or excluded based on the NHD
FTYPE.
Lakes included were FTYPEs:
• Lake/Pond
• Lake/Pond: Hydrographic Category = Perennial
• Lake/Pond: Hydrographic Category = Perennial; Stage = Average Water Elevation
• Lake/Pond: Hydrographic Category = Perennial; Stage = Date of Photography
• Lake/Pond: Hydrographic Category = Perennial; Stage = Normal Pool
• Lake/Pond: Hydrographic Category = Perennial; Stage = Spillway Elevation
• Stream/River: Hydrographic Category = Perennial
Lakes excluded were FTYPEs:
• Estuary
• Playa
• Inundation Area: Inundation Control Status = Not Controlled
• Lake/Pond: Hydrographic Category = Intermittent
• Lake/Pond: Hydrographic Category = Intermittent; Stage = Date of Photography
• Lake/Pond: Hydrographic Category = Intermittent; Stage = High Water Elevation
• Lake/Pond: Hydrographic Category = Perennial; Stage = Normal Pool
• Reservoir
• Reservoir: Construction Material = Earthen
• Reservoir: Construction Material = Nonearthen
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NLA 2022 Technical Support Document - August 2024
• Reservoir: Reservoir Type = Aquaculture
• Reservoir: Reservoir Type = Cooling Pond
• Reservoir: Reservoir Type = Decorative Pool
• Reservoir: Reservoir Type = Disposal
• Reservoir: Reservoir Type = Disposal; Construction Material = Earthen
• Reservoir: Reservoir Type = Disposal; Construction Material = Nonearthen
• Reservoir: Reservoir Type = Evaporator
• Reservoir: Reservoir Type = Evaporator; Construction Material = Earthen
• Reservoir: Reservoir Type = Filtration Pond
• Reservoir: Reservoir Type = Settling Pond
• Reservoir: Reservoir Type = Sewage Treatment Pond
• Reservoir: Reservoir Type = Tailings Pond
• Reservoir: Reservoir Type = Tailings Pond; Construction Material = Earthen
• Reservoir: Reservoir Type = Water Storage
• Reservoir: Reservoir Type = Water Storage; Construction Material = Earthen; Hyd*
• Reservoir: Reservoir Type = Water Storage; Construction Material = Earthen;
• Hydrographic Category = Intermittent
• Reservoir: Reservoir Type = Water Storage; Construction Material = Earthen;
• Hydrographic Category = Perennial
• Reservoir: Reservoir Type = Water Storage; Construction Material = Nonearthen
• Reservoir: Reservoir Type = Water Storage; Hydrographic Category = Perennial
• Reservoir; Reservoir Type = Treatment
• Swamp/Marsh
• Swamp/Marsh: Hydrographic Category = Intermittent
• Swamp/Marsh: Hydrographic Category = Perennial"
Note that excluding lake objects that are coded "Reservoir" by NHD does not exclude run-of-
the-river reservoirs or constructed ponds.
This review identified 497,840 lake objects to be included in the NLA 2022 sampling frame
(Table 2-3). The number of lake objects in the sampling frame by aggregated ecoregions and
lake are presented in Table 2-4.
Table 2-3 Number of waterbody objects in NHDPIusHR by type and sampling frame inclusion.
FTYPE
Exclude
Include
Total
LakePond
4,838,144
466,697
5,304,841
Reservoir
249,155
31,143
280,298
Estuary
8,592
0
8,592
Ice Mass
7,802
0
7,802
Playa
17,768
0
17,768
SwampMarsh
893,153
0
893,153
Total
6,014,614
497,840
6,512,454
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Table 2-4 Number of lake objects in the sampling frame by aggregated ecoregion and lake area category.
Aggregated Ecoregion
l-4ha
4-10ha
10-50ha
>50ha
Total
Coastal Plains
122,756
25,436
13,004
2,965
164,153
Northern Appalachians
20,929
6,147
4,748
1,973
33,797
Northern Plains
24,520
4,436
2,339
668
31,963
Southern Appalachians
39,413
5,201
2,346
755
47,714
Temperate Plains
54,274
10,853
6,410
1,859
73,406
Upper Midwest
30,928
10,963
9824
4,052
55,767
Western Mountains
17,319
4,963
2,880
993
26,155
Xeric
11,330
2,775
1,907
825
16,837
Total
359,553
77,086
46,430
14,771
497,840
2.3 Survey design implementation and analysis
Field crews evaluated lakes from the NLA survey design using a variety of techniques including
aerial photo interpretations, GIS analyses, local knowledge, etc. to identify lakes selected from
the sampling frame that did not meet the definition of a lake for NLA. Crews also dropped lakes
from sampling during field reconnaissance if they were a non-target type or could not be
assessed due to accessibility issues (landowner permission, too dangerous to access, etc.).
Dropped lakes were systematically replaced from a pool of replacement ("over sample") lakes
from the survey design. This process is implemented to maintain the integrity of the survey
design and to sample lakes consistent with the original number planned in different categories.
In 2022, 3,636 lakes were evaluated by field crews.
Any statistical analysis of NLA data must incorporate information about its survey design and
implementation. The statistical analysis accounts for the stratification and unequal probability
selection by using the survey design weights. The initial survey design weights are adjusted to
account for the change in sample size due to the use of over sample lakes within the strata and
unequal probability categories, i.e., the design-as-implemented weights. The adjusted weight
represents the number of lakes that each evaluated lake represents. The sum of all adjusted
weights for lakes evaluated equals the number of lakes in the sampling frame. The subset of the
lakes that are evaluated as target lakes and sampled is used to estimate the "sampled
population" of lakes by using the design-as-implemented adjusted weights. Not all lakes
evaluated as target lakes could be sampled. To account for these lakes, a second weight
adjustment, non-response weight adjustment, is completed that enables the lakes that are
target lakes and sampled to be used to estimate the "target population" of lakes.
The statistical estimates for the NLA population estimates were completed using lake weights
(see the NLA 2022 Site Information - Data file) and the R package 'spsurvey' (Dumelle et. al.
2023). Population estimates were determined at the national level and for several
subpopulations described in Chapter 10 .
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2.4 Estimated number of the NLA lakes and implications for reporting
The number of lakes in the NLA 2022 target population is not known and must be estimated
based on the lake evaluation conducted during the implementation of the survey design. The
survey design identifies lakes for evaluation from the sampling frame, which is a subset of lake
objects in NHDPIus HR described in Section 2.2. The NLA 2022 survey design identified 6,707
lake polygons for further evaluation. The NHD information may be termed the source of the
sampling frame. Note that the subset is selected such that all lake objects in the sampling frame
is expected to include all lake objects that are in the target population and may include lake
objects that the lake evaluation determines are not in the target population. An assumption is
that the sampling frame does include all lakes in the target population.
The lake evaluation categorizes the lake objects in the sample as non-target, target-not-
sampled, target-sampled and unknown. The target-not-sampled and target-sampled categories
are used to estimate the extent of the target population. Since not all lakes that are target lakes
can be sampled, the target-sampled lakes are used to estimate the extent of the sampled
population. The sampled population conceptually is all the target lakes that could have been
sampled if they were selected. The difference between the target population and sampled
population is due to "non-response" for target lakes that could not be sampled.
The initial survey design results in a survey weight for each lake that assumes that only lakes
selected to be sampled are evaluated and sampled. Since some lakes selected to be sampled
turn out to be non-target lakes or target lakes that cannot be sampled, additional lakes must be
evaluated to achieve the sample size required for each state. The initial weights are adjusted
for the survey design as implemented, i.e., the additional lakes evaluated. This initial lake
weight adjustment results in weights that may be used to estimate the extent of the lake
population and the characteristics of the sampled population. In 2007 and 2012, these weights
for the design as implemented were used for population estimates. The sampled population
estimates lead to inappropriate assumptions about the survey results (e.g., the assumption that
target lakes that could not be sampled are missing completely at random). For 2017 and 2022,
EPA determined it was more appropriate to do a second weight adjustment so that the weights
reflect the complete estimated target population. This weight adjustment accounts for the
"non-response", i.e., target lakes that could not be sampled. The weight adjustment assumes
that target lakes that could not be sampled are missing at random within the weight
adjustment categories based on the combination of state and lake area categories. See
Appendix B for a summary of the NLA survey design characteristics and estimated extent for all
four surveys.
Figure 2.1 shows the known number of lake objects in the sampling frame, the number of lakes
evaluated, and the number of lakes sampled to represent the estimated target population of
268,018. Note that to estimate the target population requires assumptions to be made about
the target lakes in the sample that could not be sampled. It is assumed that within a state, that
lakes in the same aggregated ecoregion and lake area category that could not be sampled
would have characteristics similar to those lakes that could be sampled.
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NLA 2022 Technical Support Document-August 2024
NLA 2022
Step 3: Field crews collected data from 981 randomly
selected lakes to represent the estimated 268,018
lakes in the parget population. Percentages and
confidence intervals reported for a given indicator are
relative to the target population.
Example: If EPA estimates that between 10% and 20% of
lakes are in the "most disturbed" condition for an
indicator nationally, EPA is confident that between 26,802
and 53,604 lakes nationwide are in this condition.
497,843 lakes objects in sampling frame
Vs/ Series "Samolina Frame" Point "Step 1"
Value: 497,843
3,636 randomly selected lakes were evaluated to estimate the target population
I111P
494,207 remaining lakes in the sampling frame
981 lakes were sampled to represent the estimated 268,018 lakes in the target population
267,037 unsampled target lakes
229,825 non-target lakes
0 50,000 100,000 150,000 200,000 250,000 300,000 350,000 400,000 450,000 500,000
Figure 2.1. The number of lake objects in the NLA 2022 sampling frame, evaluated lakes, sampled lakes and lakes in
the NLA target population.
2.5 Literature cited
Diaz-Ramos, S., D. L. Stevens Jr, arid A. R. Olsen. 1996. EMAP Statistical Methods Manual. US
Environmental Protection Agency, Office of Research and Development, NHEERL-
Western Ecology Division, Corvallis, Oregon.
Dumelle, M., T. Kincaid, A.R. Olsen, and M. Weber. 2023. Spsurvey: Spatial Sampling Desing and
Analysis in R. Journal of Statistical Software, 105(3), 1-29.
https://doi.org/10.18637/iss.vl05.i03
Olsen, A. R., T. M. Kincaid and Q. Payton (2012). Spatially balanced survey designs for natural
resources. Design and Analysis of Long-Term Ecological Monitoring Studies. R. A. Gitzen,
J. J. Millspaugh, A. B. Cooper and D. S. Licht. Cambridge, UK, Cambridge University Press:
126-150.
Omernik, J. M. 1987. Ecoregions of the Conterminous United States. Annals of the Association
of American Geographers. 77:118-125.
R Core Team. 2013. R: A language and environment for statistical computing. R Foundation for
Statistical Computing, Vienna, Austria. http://www.R-proiect.org.
Stevens, D. L., Jr., and A. R. Olsen. 2003, Variance estimation for spatially balanced samples of
environmental resources. Environmetrics. 14:593-610.
Stevens, D. L., Jr., and A. R. Olsen. 2004. Spatially-balanced sampling of natural resources.
Journal of American Statistical Association. 99:262-278.
Step 1: EPA found 497,843 lake objects in the
National Hydrography Dataset (NHDPIusHR)
that met eligibility criteria for inclusion in the
NLA sampling frame.
Step 2: Field crews evaluated 3,636 randomly
selected lakes. These were used to estimate the
target population of 268,018. Sampling eligibility
criteria included:
• Surface area > 1 hectare
• Depth > 1 meter
• Open water > 0.1 hectare
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NLA 2022 Technical Support Document - August 2024
USEPA. 2011. Level III Ecoregions of the Continental United States (revision of Omernik, 1987).
US Environmental Protection Agency, National Health and Environmental Effects
Laboratory Western Ecology Division. Corvallis, Oregon.
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Chapter 3: Defining Reference Sites and Condition
3.1 Background information
NLA analysts used two types of benchmarks for determining condition estimates (good, fair,
poor; above/below benchmark, etc) in the NLA public report. For trophic status, recreational
indicator microcystin, dissolved oxygen, and atrazine, analysts used fixed, nationally consistent
benchmarks that are discussed in Chapter 6 of this document. The second approach was to
establish regionally consistent reference-based benchmarks.
Reference sites are those locations that display the best available (or least-disturbed) chemical,
physical, and biological habitat condition given the current state of the landscape. To identify
these sites, data from proposed sites were compared to a definition of what is least disturbed
by human activities. To reflect the natural variability of the U.S., the definition of what is least
disturbed varies by ecological region (ecoregion). The approach used in the NLA for developing
benchmarks using reference conditions is consistent with current science, EPA guidance, state
practice, and established protocols for ecological assessment (Bailey et al., 2004; Barbour et.al.,
1999; Carter and Resh, 2013; Hughes, 1995; Reynoldson et.al., 1997; Stoddard et.al., 2006; and
USEPA, 2011).
The EPA's approach for establishing reference conditions in the NLA is a well-documented,
systematic process that screens sites using chemical and physical data to identify the least
disturbed sites within each ecological region. The application of percentiles for selecting
benchmarks is also consistent with established guidance and practice within the scientific
community and state programs (Arizona DEQ, 2012; Vermont DEC, 2016; USEPA Case Studies).
The specific approaches used in the NLA have been used in various water quality surveys since
the early 1990s and in the scientific literature since the mid-1990s (US EPA, 1998; Barbour et
al., 1999; Gerritsen, 1995; Stoddard et al., 2006; and Herlihy et al., 2008). The reference-based
approach is used by many organizations for defining benchmarks for assessing water quality.
Related to nutrients, EPA's guidance for development of nutrient criteria includes identification
of reference reaches considered to be the least impacted systems of the ecological region and
recommends the 75th percentile of the nutrient reference condition distribution for selecting a
criterion (USEPA 2000). Detailed information on the regionally consistent approach is presented
below. A summary of all benchmarks used to generate the condition estimates in the public
report can be found in Appendix C.
In refining benchmarks for NLA 2017, some 2012 benchmark values were updated; therefore,
direct comparisons should not be made between 2017 and 2022 reported results and the
results in 2007 and 2012 reports, as this will produce erroneous results. For purposes of
identifying change in this document and the public report, prior results were recalculated based
on new benchmarks as needed.
To assess ecological condition, it is standard scientific practice to compare measurements to
reference condition. The NLA approach for identifying reference sites is more inclusive than
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NLA 2022 Technical Support Document - August 2024
some approaches that restrict reference sites to only those with no or minimal human
modification; or historical, pre-industrial or pre-Columbian conditions. Because of this,
reference sites for this analysis are more accurately described as "least disturbed sites." Least
disturbed sites contain the best available chemical, physical, and biological habitat conditions
given the current state of the landscape - or "the best of what's left" (Stoddard et al. 2006).
Benchmarks were based on the distribution or range of values found for each indicator at the
reference sites (or sites with the best available conditions given today's state of the landscape)
in each of nine major ecoregions. A total of four sets of reference sites were developed for use
in establishing reference condition for the NLA results: one for the benthic macroinvertebrate
indicator, one for the zooplankton indicator, one for the nutrient indicators, and one for the
physical habitat indicators. This section describes the selection of the biological reference sites,
which also form the basis for all the nutrient and habitat reference sites.
3.2 Pre-sampling screening (hand-picked sites only)
In addition to the probability set of lakes, a smaller set of sites were hand selected a priori for
sampling. We were trying to ensure that we captured samples from additional least disturbed
lakes. Potential hand-picked sites were identified as high-quality sites by EPA, states, tribes, and
federal partners. When data were available, these potential sites were compared to water
quality screens. When data were not available, sites underwent a high-level visual screen. The
screen was used to minimize human disturbance around potential lakes (Herlihy et al., 2013).
We identified 91 hand-picked lakes for sampling following this coarse screening process. The
hand-picked sites were sampled during the 2017 index period using NLA sampling protocols,
samples were processed and analyzed with the same analytical methods as the probability site
samples, and then both the hand-picked sites and the probability sites were subjected to the
post-sample screening process (Section 3.3). Regardless of whether sites were probability-
based or hand-selected, only those that met the final screening criteria for the appropriate
indicator (i.e., benthic macroinvertebrates, zooplankton, nutrients, and physical habitat) were
used in developing reference conditions. Reference site classification and screening was done
using the nine aggregate NARS ecoregions (Figure 3-1).
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Ecoregions used In National Aquatic Resource Surveys
Figure 3.1. Nine aggregate ecoregions used for reference site classification.
3.3 Post-sampling screening for biological reference condition
To maximize the number of reference sites available for data analysis, hand-selected and
probability-based sampled in either NLA 2007, 2012 or 2017 were considered potential
reference lakes. Analysts used the chemical and physical data collected at each site to
determine whether any given site was in least disturbed condition for its aggregate ecoregion
following the approach described by Herlihy et al. (2008). The nine aggregate NARS ecoregions
were used for the ecoregion classification although in some cases these ecoregions were
further combined or lake types (natural vs. human-made) within an ecoregion were treated
differently (Figure 3-1). In the NLA, screening values were established for twelve chemical and
physical parameters to screen for biological reference sites (Table 3-1). If measurements at a
site exceeded the screening value for any one stressor, it was dropped from reference
consideration. Given that expectations of least disturbed condition vary across regions, the
criteria values for exclusion varied by ecoregion as well.
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Details on the calculation and naming of the shoreline habitat disturbance metrics is given in
the physical habitat chapter (Section 5.3). Scoring of the disturbances on the visual assessment
form for agricultural, residential, and industrial disturbance were simply done by summing the
number of checked off disturbances on the form weighting for the noted level of disturbance.
Low disturbance was weighted as 1 point, medium disturbances were weighted as 3 points, and
high disturbances were weighted as 5 points. Fire was not summed in with the industrial
disturbances as it could be an entirely natural disturbance.
All selected lake reference sites were also screened for excessive lake drawdown that was likely
anthropogenic. Evidence of both horizontal and vertical lake level fluctuations were recorded
by field crews. The square root of lake surface area was used as a surrogate for lake diameter
and was used to scale horizontal exposure of littoral lake bottom. Similarly, lake maximum
depth was used to scale vertical lake fluctuations. In addition, the drawdown criterion was
relaxed for lakes with elevated levels of lakeshore disturbance, as indexed by HiiALL_syn >
0.75. A step by step key to reference screening NLA lakes impacted by drawdown is provided in
Table 3-3.
Table 3-1. Least disturbed reference screening filter thresholds for NLA 2017.
If a lake exceeded any one of the thresholds it was not considered as a least disturbed reference site for that
ecoregion. Three filters were applied universally across all ecoregions, 1) ANC < 25 ueq/L and DOC < 5 mg/L, 2)
HifPany_Circa_syn& > 0.9, and 3) no excessive lake drawdown (see Table 3-3).
Aggregate
TP
TN
CI
S04
Turbidity
Hii-
Hii-
Assessment5
Ecoregion
(ug/L)
(ug/L)
(ueq/L)
(ueq/L)
(NTU)
NonAg&
Ag&
(Ag/Res/Ind)
WMT
>30®
>400
>100*
>200
>3
>0.6
>0
> 5/5/5
XER
>100
>1000
>500
>1000
>5
>1.5
>0.2
> 5/5/5
NPL
>150
>2000
>1000
—
>5
>1.5
>0.5
> 10/6/6
SPL
>150*
>2000*
>1000
—
>5
>1.5
>0.5
> 10/6/6
TPL
>120
>2000
>1000
>5000
>5.5
>1.7
>0.15
> 9/9/9
UMW
>40
>1200
>200
>200
>5
>0.6
>0
> 5/5/5
CPL
>50
>1200
>1000
>400
>5
>1.0
>0
> 6/10/6
SAP
>35
>800
>125
>300
>5
>0.9
>0
> 6/6/6
NAP
>30
>600
>100*
>300
>5
>0.6
>0
> 6/6/6
— metric not used for screening
& HiiNonAg_syn, HiiAg_syn, and HifPany_Circa_syn are lakeshore physical habitat disturbance indices
(see Section 5.3.4.6).
$ Assessment filters are based on indices of agricultural, residential, and industrial disturbance calculated from
observations on the visual assessment form.
* No nutrient (TP, TN) or Turbidity filters applied in Sand Hills in SPL (Omernik Level III Ecoregion 44)
# No Chloride filter applied in Coastal Ecoregions in NAP (ecoregions 59,82), XER (ecoregion 6), and WMT
(ecoregions 1,2,8)
@ No TP filter used in volcanic ecoregions in WMT (ecoregions 4,5,9,77)
In addition to selecting least disturbed reference sites, analysts also determined most disturbed
sites for each ecoregion. These sites were used primarily in developing biotic MMIs that would
be used in the biological assessment of the nation's lakes and in testing the strength of
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association of other indicators to anthropogenic stress. Similar to the reference lake selection
process, thresholds were used to determine which lakes were to be considered most disturbed
in each ecoregion (Table 3-2). If any site exceeded the most disturbed threshold for any one of
these screening criteria, then the site was classified as most disturbed.
Note that the NLA did not use data on land-use in the watersheds for the final reference site
screening—sites in agricultural areas (for example) may well be considered least disturbed,
provided that their chemical and physical conditions are among the least disturbed for the
region. Additionally, the NLA did not use data from the biological assemblages themselves to
define biological reference sites because the reference sites are being used to assess biological
condition and to use biological data to then define reference would constitute circular
reasoning.
Note that additional screening and refinement for macroinvertebrates, zooplankton, physical
habitat, and nutrient reference sites are described subsequently in their respective chapters.
Table 3-2. Most disturbed site screening thresholds for NLA 2017.
If a lake exceeded any one of the thresholds it was considered a most disturbed site for that ecoregion. One screen
for acidification was applied universally across all ecoregions, lakes with ANC < 0 ueq/L and DOC < 5 mg/L were
considered most disturbed.
Aggregate
TP
TN
CI
S04
Turbidity
Hii-
Hii-
Assessment5
Ecoregion
(ug/L)
(ug/L)
(ueq/L)
(ueq/L)
(NTU)
NonAg&
Ag&
(Ag/Res/Ind)
WMT
>150®
>1500
>1500*
>1500
>10
>2.5
>0.9
> 15/15/15
XER
>400
>4000
—
—
>25
>3.5
>1.0
> 15/15/15
NPL
>400
>4000
—
—
>50
>3.5
>1.2
> 15/15/15
SPL
>400*
>4000*
—
—
>50
>3.5
>1.2
> 15/15/15
TPL
>500
>5000
>5000
>20,000
>50
>4.0
>1.2
> 15/18/15
UMW
>200
>2500
>2500
>2500
>20
>3.5
>0.9
> 15/15/15
CPL
>200
>3000
>5000
>2500
>30
>3.5
>1.0
> 15/15/15
SAP
>150
>2500
>1500
>1500
>20
>3.5
>0.9
> 15/15/15
NAP
>150
>2500
>1500*
>1500
>20
>3.5
>0.9
> 15/15/15
— metric not used for screening
& HiiNonAg_syn and HiiAg_syn are lakeshore physical habitat disturbance indices (see Section 5.3.4.6)
$ Assessment filters are based on indices of agricultural, residential, and industrial disturbance calculated from
observations on the visual assessment form.
* No nutrient (TP, TN) or Turbidity filters applied in Sand Hills in SPL (Omernik Level III Ecoregion 44)
# No Chloride filter applied in Coastal Ecoregions in NAP (ecoregions 59,82), XER (ecoregion 6), and WMT
(ecoregions 1,2,8)
@ No TP filter used in volcanic ecoregions in WMT (ecoregions 4,5,9,77)
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Table 3-3. Dichotomous key for defining NLA lakes likely impacted by anthropogenic drawdown.
Based on field observations of horizontal lake level fluctuations (AH), vertical lake level
fluctuations (AV), and human lakeshore disturbance (physical habitat summary metric
HiiAII_syn).
1. AH < 10 m AND AV < 2 m
Yes - LAKE OK
No - go to 2
2. AH > 10 m and AV > 2 m
Yes - Lake Drawdown, Not Reference
No - go to 3
3. AV > 2 m and AV/Maximum Lake Depth > 10%
Yes - Lake Drawdown, Not Reference
No - go to 4
4. AH < 10 m
Yes - LAKE OK
No - go to 5
5. AH/sqrt(Lakearea) > 5%
Yes - Lake Drawdown, Not Reference
No - go to 6
6. Lake Disturbed, HiiAII_syn > 0.75
Yes - Lake Drawdown, Not Reference
No - LAKE OK
3.4 Literature cited
Arizona Department of Environmental Quality. 2012. Implementation Procedures for the
Narrative Biocriteria Standard, Final draft July 2012,
http://www.azdeq.gov/environ/water/standards/download/draft_bio.pdf
Bailey, R.C., R.H. Norris, and T.B. Reynoldson. 2004. Bioassessment of freshwater ecosystems:
using the reference condition approach. Kluwer Academic Publishers, New York.
Barbour, M.T., J. Gerritsen, B.D. Snyder, and J.B. Stribling. 1999. Rapid bioassessment protocols
for use in streams and wadeable rivers: periphyton, benthic macroinvertebrates and
fish, second edition. EPA 841-B-99-002. United States Environmental Protection Agency;
Office of Water. Washington, D.C.
Carter, J.L., and Resh, V.H. 2013, Analytical approaches used in stream benthic
macroinvertebrate biomonitoring programs of State agencies in the United States: U.S.
Geological Survey Open-File Report 2013-1129, 50 p.,
http://pubs.usgs.gov/of/2013/1129/
Gerritsen, J. 1995. Additive biological indices for resource management. J. North Am. Benthol.
Soc. 14(3):451-457.
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NLA 2022 Technical Support Document - August 2024
Herlihy, A. T., S. G. Paulsen, J. Van Sickle, J. L. Stoddard, C. P. Hawkins, and L. L. Yuan. 2008.
Striving for consistency in a national assessment: the challenges of applying a reference
condition approach at a continental scale. Journal of the North American Benthological
Society 27:860-877.
Herlihy, A. T., J. B. Sobota, T. C. McDonnell, T. J. Sullivan, S. Lehmann, and E. Tarquinio. 2013. An
a priori process for selecting candidate reference lakes for a national survey.
Freshwater Science 32:385-396. doi: 10.1899/11-081.1.
Hughes, R.M. 1995. Defining acceptable biological status by comparing with reference
conditions. Chapter 4 in Biological assessment and criteria: tools for water resource
planning and decision making, W.S. Davis and T.P. Simon, eds. (pp. 31 - 47). CRC Press,
Boca Raton.
Reynoldson, TB., R.H. Norris, V.H. Resh, K.E. Day, and D.M. Rosenberg. 1997. The reference
condition: a comparison of multimetric and multivariate approaches to assess water-
quality impairment using benthic macroinvertebrates. Journal of the North American
Benthological Society 16:833-852.
Stoddard, J. L., D. P. Larsen, C. P. Hawkins, R. K. Johnson, and R. H. Norris. 2006. Setting
expectations for the ecological condition of running waters: the concept of reference
condition. Ecological Applications 16:1267-1276.
USEPA, Case Studies- Setting Ecologically-Based Water Quality Goals Ohio's Tiered Aquatic Life
Use Designations Turn 20 Years Old (website). United States Environmental Protection
Agency, Office of Water, Office of Science and Technology.
http://water.epa.gov/scitech/swguidance/standards/criteria/aqlife/biocriteria/aquaticlif
eohio.cfm
USEPA. 1998. Lake and Reservoir Bioassessment and Biocriteria. Technical Guidance Document.
USEPA, Office of Wetlands, Oceans and Watershed and Office of Science and
Technology. EPA-841-B98-007. https://www.epa.gov/sites/default/files/2019-
03/documents/la ke-reservoir-tech-guidance-doc-1998.pdf
USEPA 2000. Nutrient Criteria Technical Guidance Manual. Lakes and Reservoirs. First Edition.
EPA-822-B00-001. US EPA, Office of Water, Office of Science and Technology.
https://www.epa.gov/nutrient-policy-data/nutrient-criteria-development-document-
lakes-and-reservoirs
USEPA. 2011. A Primer on Using Biological Assessments to Support Water Quality Management.
USEPA, Office of Science and Tehcnology. EPA 810-R-11-01.
https://www.epa.gov/sites/default/files/2018-10/documents/primer-using-biological-
assessments.pdf
Vermont Department of Environmental Conservation. 2016. Nutrient Criteria for Vermont's
Inland Lakes and Wadable Streams. Technical Support Document. Vermont Department
of Environmental Conservation, Watershed Management Division.
https://dec.vermont.gov/sites/dec/files/wsm/Laws-Regulations-Rules/2016_12_22-
Nutrient_criteria_technical_support_document.pdf
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Chapter 4: Benthic Macroinvertebrates
4.1 Background information
The taxonomic composition and relative abundance of different taxa that make up the littoral
macroinvertebrate assemblage present in a lake can be used to assess how human activities
affect ecological condition. Two principal types of ecological assessment tools to assess
condition based on macroinvertebrate assemblages are currently prevalent: multimetric indices
and predictive models of taxa richness. The purpose of these indicators is to present the
complex community taxonomic data represented within an assemblage in a way that is
understandable and informative to resource managers and the public. For NLA 2012, we
developed a multimetric index of macroinvertebrate condition using 2007 and 2012 NLA data
as described in Section 4.3. This NLA 2012 MMI and its condition class benchmarks (Table 4-3)
were used for the 2017 and 2022 macroinvertebrate assessments.
Multimetric indicators have been used in the U.S. to assess condition based on fish and
macroinvertebrate assemblage data (e.g., Karr and Chu, 2000; Barbour et al., 1999; Barbour et
al., 1995). The multimetric approach involves summarizing various assemblage attributes (e.g.,
composition, tolerance to disturbance, trophic and habitat preferences) as individual "metrics"
or measures of the biological community. Candidate metrics are then evaluated for various
aspects of performance and a subset of the best performing metrics are then combined into an
index, referred to as a multimetric index or MMI.
4.2 Data preparation
4.2.1 Standardizing counts
The number of individuals counted in a sample was standardized to a constant number to
provide an adequate number of individuals that was the same for the most samples and that
could be used for multimetric index development. A subsampling technique involving random
sampling without replacement was used to extract, from the dataset, a true "fixed count" of
300 individuals from the total number of individuals enumerated for a sample (target lab count
was 500 individuals). Samples that did not contain at least 300 individuals were used in the
assessment because low counts can indicate a response to one or more stressors.
4.2.2 Autecological characteristics
Autecological characteristics refer to specific ecological requirements or preferences of a taxon
for habitat preference, feeding behavior, and tolerance to human disturbance. These
characteristics are prerequisites for identifying and calculating many metrics. A number of
state/regional organizations and research centers have developed autecological characteristics
for benthic macroinvertebrates in their region. For the NLA, a consistent "national" list of
characteristics that consolidated and reconciled any discrepancies among the regional lists was
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needed before certain biological metrics could be developed and calibrated and an MMI could
be constructed. The same autecological information used in WSA and NRSA was used in NLA.
Members of the data analysis group pulled together autecological information from five
existing sources: the EPA Rapid Bioassessment Protocols document, the National Ambient
Water Quality Assessment (NAWQA) national and northwest lists, the Utah State University list,
and the EMAP Mid-Atlantic Highlands (MAHA) and Mid-Atlantic Integrated Assessment (MAIA)
list. These five were chosen because they were thought to be the most independent of each
other and the most inclusive. A single national-level list was developed based on the following
decision rules for tolerance values, functional feeding group and habitat preferences, and
taxonomic resolution.
4.2.3 Tolerance values
Tolerance value assignments followed the convention for macroinvertebrates, ranging between
0 (least tolerant or most sensitive) and 10 (most tolerant). For each taxon, tolerance values
from all five sources were reviewed and a final assignment made according to the following
rules:
1. If values from different lists were all <3 (sensitive), final value = mean;
2. If values from different lists were all >3 and <7 (facultative), final value = mean;
3. If values from different lists were all >7 (tolerant), final value = mean;
4. If values from different lists spanned sensitive, facultative, and tolerant categories,
best professional judgment was used, along with alternative sources of information
(if available) to assign a final tolerance value; and
5. Tolerance values of 0 to <3 were considered "sensitive" or "intolerant." Tolerance
values >7 to 10 were considered "tolerant," and values in between were considered
"facultative."
4.2.4 Functional feeding group and habitat preferences
In many cases, there was agreement among the five data sources. When discrepancies in
functional feeding group (FFG) or habitat preference ("habit") assignments among the five
primary data sources were identified, a final assignment was made based on the most
prevalent assignment. In cases where there was no prevalent assignment, the workgroup
examined why disagreements existed, flagged the taxon, and used best professional judgment
to make the final assignment.
4.2.5 Taxonomic resolution
Taxonomic resolution is an important factor in the development of multimetric indices.
Maintaining consistent taxonomic resolution for specific taxa across sites helps ensure that
differences between sites are due to environmental factors and not an artifact of taxa
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identifications. For most taxa identified, the taxonomic resolution was to the generic level,
however the following groups had higher-level hierarchical taxonomic resolution: oligochaetes,
mites, polychaetes were rolled up to family, ceratopogonids were rolled up to subfamily.
4.3 Multimetric index development
4.3.1 Dataset
The NLA macroinvertebrate 300 fixed count data were used to calculate the community metrics
used in the MMI. A best ecoregional MMI was developed by scoring and summing the six
metrics that performed best in each ecoregion. The NLA macroinvertebrate MMI was
developed using the combined the NLA 2007 and 2012 benthic metric files which were both
calculated with common autecology and taxonomic resolution. All reference sites were defined
using the NLA definitions described in Section 3 based on nine aggregate ecoregion criteria.
Reference sites that had less than 250 individuals were not used as reference for MMI
development. Altogether, there were 2330 site visits (samples) in the data used to develop the
MMI; 1132 from 2007 and 1198 from 2012. There were 1789 unique sites. Some sites were
sampled twice in their respective years and some sites were sampled in both 2007 and 2012.
4.3.2 Low macroinvertebrate numbers
Many samples had a very low number of individuals. Examination of these low number sites did
not suggest that this was primarily due to impairment. We think that it is related to field
collection and lake bottom substrate composition. Samples with low bug numbers will have
poor MMI scores because of the strong relationship between sample count and taxa richness.
We decided that samples with less than 100 individuals were not sufficiently sampled and we
would not assess them. They were removed from the process of MMI development and MMI
scores for them will be set to missing values. These are identified as "not assessed" for
macroinvertebrates in the NLA. In the NLA 2017 data, 60 of the 1191 samples had < 100
individuals. In NLA 2022, 55 of the 1071 samples had <100 individuals.
4.3.3 Ecoregion classification
For the NLA macroinvertebrate MMI development, the nine national aggregate ecoregions
(Figure 3-1) were consolidated into five aggregate biological ecoregions by combining some
ecoregions together. Specifically, that consisted of making an Eastern Highlands (EHIGH) region
by combining the SAP and NAP, a PLAINS ecoregion by combining the TPL, SPL, and NPL, and a
Western ecoregion (WMTNS) by combing the WMT and XER regions. The CPL and UMW remain
their own ecoregions. MMIs were developed independently for each of these 5 biological
ecoregions.
4.3.4 Metric screening
All 126 calculated benthic metrics were screened for both signal:noise (S:N) and discrimination
of least disturbed reference sites from most disturbed sites (F-test). S:N ratios were calculated
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for each metric nationally and within each biological ecoregion using the visit 1 versus visit 2
variance within year as the noise and among site variance as the signal. For calculating F-tests,
and all subsequent MMI development, we only used one visit per site (index visit). The first
sample visit of the year with valid data was used. For sites with valid samples in both years, the
2012 first visit data were used (samples with less than 100 bugs were not considered valid
data). F-tests were run on just the least disturbed reference (L) versus the most disturbed (M)
sites.
Metrics had to pass both F and S:N screens in order to remain in consideration for inclusion in
the final MMI. Metrics had to have S:N > 1.5 either nationally or within their ecoregion in order
to pass. For the F-test, only metrics that had F-values > 4.0 passed. From this screening, 35
metrics from CPL, 42 from EHIGH, 44 from UMW, 29 from PLAINS, and 50 from WMTNS passed
and were considered for the all subsets MMI selection.
4.3.5 All Subsets MMI selection
Passing metrics were assigned to one of the six basic metric classes used to assemble the MMI
as done in the NARS stream MMI (Stoddard et al., 2008). An all subsets procedure was used to
assemble all possible combinations of MMIs using the six metric class framework. There were
8,960 combinations of metrics in the CPL, 12,096 in the EHIGH, 36,855 in the UMW, 3360 in the
PLAINS, and 65,280 in the WMTNs. For each possible MMI combination, the MMI S:N, F-test,
metric correlations, and IQR box delta (separation between least and most disturbed) were
calculated. For correlations, both the mean and maximum correlation among the six metrics
were calculated. IQR box delta or separation is the difference between the 25th percentile of
reference sites and the 75th percentile of most disturbed sites. Thus, positive box deltas
indicate separation between the least and most disturbed boxes, negative values indicate
overlap in the IQRs (boxes of box and whisker plot) of the least and most disturbed sites.
To pick the best MMI from the all subsets results, all MMI candidates were first screened for
S:N and maximum metric correlation. Only MMIs that had max correlation < 0.7 and S:N > 3
were considered. MMIs that passed this screen were evaluated for both box delta and F-value
with the goal of picking the MMI that had the best combination of those two values. These two
measures are highly correlated. To do this objectively, we ran a PCA on box delta and F-value
and selected the MMI that had the highest PCA factor 1 score. The intent was to optimize and
pick the model with the best combination of F-value and separation. The six metrics that make
up the final (best) MMI are shown in Table 4-1.
Each of the six selected metrics were scored on a 0-10 scale by interpolating metrics between a
floor and ceiling value. The six metric 0-10 point scaled scores were then summed and
normalized to a 0-100 scale by multiplying by 10/6 to calculate the final MMI. Details of
this process are described in Stoddard et al. (2008) for the NARS stream MMI but the NLA
process is the same. The final metrics used in each ecoregion, metric direction, and floor and
ceiling values are summarized in Table 4-1. Scoring equations are different depending on if the
metric responds positively (high values good) or negatively (high values bad) with disturbance.
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For positive metrics, values above the ceiling get 10 points, and values below the floor get 0
points. For negative metrics, values above the ceiling get 0 points, and values below the floor
get 10 points. The interpolation equations for scoring the 0-10 points for metrics between the
floor and ceiling values are:
Positive Metrics: Metric Points = 10*((metric value-floor)/(ceiling-floor)); and
Negative Metrics: Metric Points = 10 * (1 - ((metric value-floor)/(ceiling-floor))).
For positive metrics, floor values are set at the 5th percentile of all samples in the ecoregion,
ceiling values are the 95th percentile of reference sites in the ecoregion. Negative metric
floor/ceilings are calculated the opposite way. Statistics for the final MMI in each ecoregion are
shown in Table 4-2. The overall S:N of the MMI based on visit 1 vs. 2 revisits nationally across
both years was 3.56. Box plots showing the R versus T discrimination of the final MMIs are
shown in Figure 4-1.
Table 4-1. Final NLA biological ecoregion benthic MMI metrics and their floor/ceiling values for MMI scoring.
Ecoregion
Metric Class
Metric name*
Direction
Floor
Value
Ceiling
Value
Coastal Plains
Composition
NOINPTAX
Negative
21.88
55.17
Coastal Plains
Diversity
CHIRDOM5PIND
Negative
55.71
100.0
Coastal Plains
Feeding Group
PREDNTAX
Positive
6.00
23.0
Coastal Plains
Habit
SPWLNTAX
Positive
5.00
15.0
Coastal Plains
Richness
EPT_NTAX
Positive
1.00
8.00
Coastal Plains
Tolerance
NTOLPIND
Positive
6.33
64.33
E. Highlands
Composition
NOINPTAX
Negative
13.79
48.72
E. Highlands
Diversity
CHIRDOM5PIND
Negative
57.46
95.24
E. Highlands
Feeding Group
COGANTAX
Positive
8.00
27.0
E. Highlands
Habit
CLNGNTAX
Positive
3.00
12.0
E. Highlands
Richness
EPOTNTAX
Positive
2.00
14.0
E. Highlands
Tolerance
TL23NTAX
Positive
1.00
9.00
Plains
Composition
DIPTPTAX
Negative
16.67
60.00
Plains
Diversity
CHIRDOM5PIND
Negative
50.44
100.0
Plains
Feeding Group
PREDNTAX
Positive
2.00
19.0
Plains
Habit
CLMBPTAX
Positive
10.0
33.33
Plains
Richness
EPOTNTAX
Positive
0
10.0
Plains
Tolerance
TL23PIND
Positive
0
19.67
Upper Midwest
Composition
NOINPIND
Negative
5.33
89.0
Upper Midwest
Diversity
CHIRDOM3PIND
Negative
36.51
89.29
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Ecoregion
Metric Class
Metric name*
Direction
Floor
Value
Ceiling
Value
Upper Midwest
Feeding Group
SHRDPIND
Negative
2.67
50.67
Upper Midwest
Habit
CLNGNTAX
Positive
3.00
14.0
Upper Midwest
Richness
CRUSNTAX
Negative
0
3.00
Upper Midwest
Tolerance
TL23PTAX
Positive
2.17
23.81
Western Mts.
Composition
DIPTPIND
Positive
5.97
84.33
Western Mts.
Diversity
HPRIME
Positive
1.09
2.87
Western Mts.
Feeding Group
SCRPNTAX
Negative
0
5.00
Western Mts.
Habit
CLNGNTAX
Positive
1.00
8.00
Western Mts.
Richness
EPT_NTAX
Positive
0
7.00
Western Mts.
Tolerance
TL23PTAX
Positive
0
21.43
*Metric Names
NOINPTAX=% Non-Insect Taxa (Non-Insect Taxa Richness /Total Taxa Richness*100)
DIPTPTAX = % Diptera Taxa (Diptera Taxa Richness / Total Taxa Richness*100)
NOINPIND = % Non-Insect Individuals
ODONPIND = % Odonata Individuals
CHIRDOM3PIND = % Chironomid Individuals in Top 3 most abundant Chironomid Taxa
CHIRDOM5PIND = % Chironomid Individuals in Top 5 most abundant Chironomid Taxa
HPRIME = Shannon Diversity Index
PREDNTAX = Predator Taxa Richness
COGANTAX = Collector-Gatherer Taxa Richness
SHRDPIND = % Shredder Individuals
SCRPNTAX = Scraper Taxa Richness
SPWLNTAX = Sprawler Taxa Richness
CLNGNTAX = Clinger Taxa Richness
CLMBPTAX = % Climber Taxa (Climber Taxa Richness/Total Taxa Richness *100)
EPT_NTAX = Ephemeroptera + Plecoptera + Trichoptera Taxa Richness
EPOTNTAX = Ephemeroptera + Plecoptera + Trichoptera + Odonata Taxa Richness
CRUSNTAX = Crustacean Taxa Richness
TRICNTAX = Trichoptera Taxa Richness
NTOLPIND = % Individuals with pollutant tolerance values < 6
TL23NTAX= Taxa Richness of taxa with pollutant tolerance values > 2.0 and < 4.0
TL23PIND = % Individuals with pollutant tolerance values > 2.0 and < 4.0
TL23PTAX = % Taxa with pollutant tolerance values > 2.0 and < 4.0
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Table 4-2. Benthic MM! statistics for the NLA 2007-2012 data used to develop the MML
Ecoregion
F-test
Box Delta
Max Corr.
Mean Corr.
S:N
Coastai Plains
54.7
12.7
0.45
0.17
3.45
E. Highlands
69.0
1.85
0.50
0.26
3.12
Plains
36.2
-2.26
0.68
0.41
3.35
Upper Midwest
64.5
10.4
0.57
0.24
3.00
Western Mts.
88.9
4.46
0.48
0.16
3.66
F-test=F-score for difference between least disturbed (reference) and most disturbed site means; Box
Delta=Separation difference between Reference Q1 and most disturbed Q3 in MMI units; Corr=Pearson correlation
among six MMI metrics; S:N = Ecoregional within year S:N ratio.
u
q-
¦1
|
II
[
l
I
II
II
L
M
CPL
WMTNS
EHIGH PLAINS UMW
Aggregate Ecoregions
Figure 4,1. Box and whisker plots showing discrimination between least disturbed reference (L) and most disturbed
(M) sites by biological ecoregion in the NLA 2007-2012 data used to develop the MMI. Boxes show the
interquartile range and the whiskers show the 5th and 95th percentiles. Outliers are not presented.
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4.3.6 Setting MMI benchmarks
Previous large-scale assessments have converted MMI scores into classes of assemblage
condition by comparing those scores to the distribution of scores observed at least disturbed
reference sites. See Section 3.3 for information on selecting reference sites. If a site's MMI
score was less than the 5th percentile of the reference distribution, it was classified as in most
disturbed condition; scores between the 5th and 25th percentile were classified as moderately
disturbed and scores in the 25th percentile or higher were classified as least disturbed.
For calculating the benchmarks used in the NLA 2022 public report, we used all NLA reference
sites sampled from 2007-2017 to maximize sample sizes used to calculate percentiles. When a
site was sampled multiple times, only the first visit to the most recent year of sampling was
used to calculate percentiles so sites were not double-counted. Also, only reference sites with
at least 250 individuals were used. Before calculating benchmarks, a 1.5*IQR outlier analysis
was done on the reference site MMIs to remove outliers. No sites were dropped as outliers in
this process leaving 416 reference sites for calculating reference site percentiles to use as
benchmarks. The resulting adjusted MMI benchmark values for the condition classes in each
ecoregion are given in Table 4-3.
Table 4-3. Macroinvertebrate MMI benchmarks using 2007-2017 reference site data
Ecoregion
#of Ref Sites
Least Disturbed
25th Percentile Benchmark
Most Disturbed
5th Percentile Benchmark
Coastal Plains
29
>51.8
<40.4
East. Highlands
105
>44.5
<31.4
Plains
84
>39.5
<26.6
Upper Midwest
76
>51.4
<37.2
Western Mountains
122
>47.6
<32.6
4.4 Literature cited
Barbour, M. T., J. Gerritsen, B. D. Snyder, and J. B. Stribling. 1999. Rapid bioassessment
protocols for use in streams and wadeable rivers. EPA 841/B-99/002. Office of Water,
US Environmental Protection Agency, Washington, DC.
Barbour, M. T., J. B. Stribling, and J. R. Karr. 1995. Multimetric approach for establishing
biocriteria and measuring biological condition. Pages 63-77 in W. S. Davis and T. P.
Simon (editors). Biological assessment and criteria: tools for water resource planning
and decision making. Lewis Publishers, Boca Raton, Florida.
Herlihy, A. T., S. G. Paulsen, J. Van Sickle, J. L. Stoddard, C. P. Hawkins, and L. L. Yuan. 2008.
Striving for consistency in a national assessment: the challenges of applying a reference
condition approach at a continental scale. Journal of the North American Benthological
Society 27:860-877.
Karr, J. R., and E. W. Chu. 2000. Sustaining living rivers. Hydrobiologia 422/423:1-14.
Stoddard, J. L., A. T. Herlihy, D. V. Peck, R. M. Hughes, T. R. Whittier, and E. Tarquinio. 2008. A
process for creating multi-metric indices for large scale aquatic surveys. Journal of the
North American Benthological Society 27:878-891.
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Chapter 5:
NLA 2022 Technical Support Document
Physical Habitat
-August 2024
5.1 Background information
Near-shore physical habitat structure in lakes has only recently been addressed by the U.S.
Environmental Protection Agency (EPA) in its National Aquatic Resource Surveys (NARS)
monitoring efforts (e.g., USEPA 2009, Kaufmann et al. 2014a,b,c). Like human activities, aquatic
and riparian biota are concentrated near lakeshores, making near-shore physical habitat
ecologically important, but exposed and vulnerable to anthropogenic perturbation (Schindler
and Scheuerell 2002, Strayer and Findlay 2010, Hampton et al. 2011). Littoral and riparian zones
are positioned at the land-water interface and tend to be more structurally complex and
biologically diverse than either pelagic areas or upland terrestrial environments (Polis et al.
1997, Strayer and Findlay 2010). This complexity promotes interchange of water, nutrients, and
biota between the aquatic and terrestrial compartments of lake ecosystems (Benson and
Magnuson 1992, Polis et al. 1997, Palmer et al. 2000, Zohary and Ostrovsky 2011). Structural
complexity and variety of cover elements in littoral areas provide diverse opportunities for
supporting assemblages of aquatic organisms (Strayer and Finlay 2010; Kovalenko et al 2012),
while intact riparian vegetation and wetlands surrounding lakes increase near-shore physical
habitat complexity (e.g., Christensen et al. 1996, Francis and Schindler 2006) and buffer lakes
from the influence of upland land use activities (Carpenter and Cottingham 1997, Strayer and
Findlay 2010). Human activities on or near lakeshores can directly or indirectly degrade littoral
and riparian habitat (Francis and Schindler 2006). Increased sedimentation, loss of native plant
growth, alteration of native plant communities, loss of physical habitat structure, and changes
in littoral cover and substrate are all commonly associated with lakeshore human activities
(Christensen et al. 1996, Engel and Pederson 1998, Whittier et al. 2002, Francis and Schindler
2006, Merrell et al. 2009). Such reductions in physical habitat structural complexity can
deleteriously affect fish (Wagner et al. 2006, Taillon and Fox 2004, Whittier et al. 1997, 2002,
Halliwell 2007, Jennings et al. 1999, Wagner et al. 2006), aquatic macroinvertebrates (Brauns et
al. 2007), and birds (Kaufmann et al. 2014b).
The EPA developed standardized, rapid field methods to quantify physical habitat structure and
near-shore anthropogenic disturbances (Kaufmann and Whittier 1997) and piloted them in the
Northeastern U.S. (Larsen and Christie 1993, Whittier et al. 2002b, Kaufmann et al. 2014b).
These methods were modified (USEPA 2007a, Kaufmann et al. 2014a) and applied in 2007 for
the first U.S. national survey of lake physical habitat condition (US EPA 2009, Kaufmann et al.
2014c). The EPA's lake physical habitat methods were once again modified to explicitly assess
habitat structure in exposed drawdown zones (USEPA 2012) and applied in the NLA 2012 survey
as part of the EPA's second national survey of the ecological condition of lakes in the United
States (USEPA 2016). The NLA 2012 field method modifications were structured so that we
were able to duplicate all the lake habitat condition indices that were used in the previous
(2007) national assessment. We calculated habitat metrics and indices described by Kaufmann
et al. 2014a,c) to quantify the variety, structural complexity, and magnitude of areal cover from
physical habitat elements within the near shore zones of lakes in the NLA 2012 survey. For the
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NLA 2017 physical habitat condition we used the same expected condition models that we used
for the 2012 Assessment, with the exception of lake drawdown that is discussed in more detail
below.
Our objectives in this chapter are to describe how we calculated physical habitat indices based
on near-shore physical habitat data collected in the NLA survey, and how we derived physical
habitat condition benchmarks relative to least disturbed conditions. We only briefly describe
the NLA field methods and data reduction procedures, which are published elsewhere (USEPA
2012; Kaufmann 2014a, USEPA 2017). Finally, we evaluate the precision of NLA's key indices of
physical habitat condition and examine their association with anthropogenic disturbances.
5.2 Data preparation
We took the following eight steps to assess physical habitat condition in U.S. lakes based on the
NLA 2017 national probability sample of lakes and reservoirs. For the NLA 2017 physical habitat
condition we used exactly the same expected condition models that we used for the 2012
Assessment, which were derived using combined NLA 2007 and NLA 2012 data, including
reference sites defined based on NLA 2012 screening criteria. [But see notes on
accommodating missing horizontal and vertical lake drawdown measurements.]
1) Field crews made measurements and observations of near-shore physical habitat structure
and human activities on a national probability sample of lakes and reservoirs (described by
USEPA 2016, and Kaufmann et al. 2014a);
2) Classified survey lakes by aggregated ecoregion (ECOWSA9_2015), and by their relative
levels of anthropogenic disturbance within those ecoregions (RT_NLA12_2015).
3) Calculated a set of physical habitat metrics as described by Kaufmann et al. (2014a) for NLA
2007, but adapted calculations to adjust for the NLA 2012's field method change that
assessed riparian vegetation cover, littoral cover, and human disturbance in the drawdown
zone separate from those above the typical high water mark or inundated by water in the
littoral zone;
4) Calculated multimetric indices of lakeshore anthropogenic disturbance and nearshore
physical habitat cover and structure as described by Kaufmann et al. (2014c) for NLA 2007,
and assigned variants of these indices according to aggregated Ecoregions
(ECOWSA9_2015); also defined a new indicator of lake drawdown;
5) Estimated lake-specific expected ("E") values for physical habitat indices from region-
specific regression models of factors predicting physical habitat in the combined set of least
disturbed lakes from the NLA 2007 and 2012 surveys. Our modeling approach is very similar
to that employed by Kaufmann et al. (2014c) in the Western Mountain and Xeric ecoregions
for the NLA 2007 report;
6) Set criteria for low, medium and high lakeshore anthropogenic disturbance (good, fair,
poor) based on professional judgement; good, fair, and poor littoral and riparian physical
habitat condition based on deviation from the central tendency of observed/expected (O/E)
values within the group of least disturbed lakes; and small, medium, and large lake
drawdown based on percentiles of the indicator values themselves in least disturbed lakes.
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7) Examined the precision of NLA 2012 key physical habitat indicators.
8) Examined the association between NLA 2012 physical habitat indicators and anthropogenic
disturbances, comparing the regional distributions of habitat condition in least disturbed
reference lakes with those in most disturbed lakes.
5.3 Methods
5.3.1 NLA sites used for expected condition modeling and precision estimates
The NLA field sampling effort targeted all lakes and reservoirs in the 48 conterminous U.S. with
surface areas >1 ha and depths greater than 1 m. Field data were collected between May and
October of each survey year. See Chapter 2 of this document for additional details on the study
area and site selection. To model expected condition for all four NLA surveys ('07, '12, '17, '22),
we used physical habitat data collected in the 2007 and 2012 survey years. These data included
data from 2268 lakes and reservoirs, 1156 in 2007, and 1112 in 2012. Probability and hand-
selected lakes from both 2012 and 2007 were used to develop expected physical habitat
condition models and distributions of O/E values in least-disturbed lakes. Random subsets of 90
probability lakes from NLA 2007 and 88 from NLA 2012 were visited twice during their
respective summer sampling periods to estimate the precision of NLA indicators, including the
habitat measurements and indices (Kaufmann et al. 2014a).
5.3.2 Field sampling design and methods
Our lake physical habitat field methods (USEPA 2007a, USEPA 2012, USEPA 2017, Kaufmann et
al. 2014a) produced information concerning 7 dimensions of near-shore physical habitat: 1)
water depth and surface characteristics, 2) substrate size and type, 3) aquatic macrophyte
cover and structure, 4) littoral cover for biota, 5) riparian vegetation cover and structure, 6)
near-shore anthropogenic disturbances, and 7) bank characteristics that indicate lake level
fluctuations and terrestrial-aquatic interactions. At each lake, field crews characterized these 7
components of near-shore physical habitat at 10 equidistant stations along the shoreline. Each
station included a littoral plot (10m x 15m) abutting the shoreline, a riparian plot (15m x 15m)
extending landward from the typical high-water mark, and in a 15m wide drawdown zone plot
that extended a variable distance landward, depending on the amount of lake level drop
compared with typical high water levels (Figure 5-1). Littoral depth was measured 10 m off-
shore at each station. Metrics and indices were calculated for the variable-width drawdown
zone plots, the 15m x 15m riparian plots and the 10m x 15m littoral plots. To match the riparian
and near-shore human disturbance indices to those used in the previous (NLA 2007)
assessment, we used information from riparian and drawdown plots along with drawdown
horizontal extent information. These index values are equivalent to the 2007 index values that
were directly calculated from observation the near-shore zone extending from the lake water's
edge 15m outward. See Kaufmann et al. (2014a) for further description of field methods, our
approach for calculating whole-lake physical habitat metrics, and a detailed assessment of
habitat metric precision.
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5.3.3 Classifications
5.3.3.1 Ecoregions
We report findings nationally, and by 9 aggregated Omernik (1987) level III ecoregions (Paulsen
et al. 2008) including the Northern Appalachians (NAP), Southern Appalachians (SAP), Coastal
Plains (CPL), Upper Midwest (UMW), Temperate Plains (TPL), Northern Plains (NPL), Southern
Plains (SPL), Western Mountains (WMT), and Xeric West (XER) (Figure 3-1). We used ecoregions
as a first-level classification for defining and evaluating near-shore riparian and littoral
condition indicators (RVegQ, LitCvrQ, and LitRipCvrQ) and their variants (e.g., RVegQ_2,
LitCvrQ_b, LitRipQ_2d). Ecoregions are useful predictors of many characteristics of landform,
geology, climate, hydrology, and potential natural vegetation (Omernik 1987, Paulsen et al.
2008) that influence physical habitat in lakes (Kaufmann et al. 2014c). Kaufmann et al. (2014c)
used a multivariate classification of lake characteristics including lake chemistry and depth to
assign variants of LitCvrQ, suggesting that such classifications would capture aspects of in-lake
habitat cover complexity better than would ecoregions. We reexamined the 2007 data and
found no substantial difference in assignment of LitCvrQ variants according to Ecoregion
(WSAEC09) versus multivariate cluster analysis (CLUSB). For some aspects of habitat index
development, we grouped ecoregions into broader ecoregions. The grouping included the
Eastern Highlands (EHIGH = NAP + SAP), the Plains and Lowlands (PLNLOW = CPL + UMW + TPL
+ NPL + SPL), Central Plains (CENPL = TPL+ NPL+SPL), and the West (WMT + XER).
5.3.3.2 Anthropogenic disturbance and least disturbed reference site screening
We used region-specific screening based on water chemistry, near-shore human influences, and
evidence of anthropogenic lake drawdown in NLA survey lakes, 1109 from NLA 2012 and 1101
from NLA 2007, to classify all NLA lakes according to their level of anthropogenic disturbance
(low, medium, high), as described in Chapter 3. Lakes meeting low-disturbance screening
criteria served as least disturbed reference sites for best-available condition. Low-disturbance
stress (least disturbed) lakes within each Ecoregion were identified on the basis of chemical
variables (total phosphorus, total nitrogen, chloride, sulfate, acid neutralizing capacity,
dissolved organic carbon, and dissolved oxygen in the epilimnion) and direct observations of
anthropogenic disturbances along the lake margin (proportion of lakeshore with non-
agricultural influences, proportion of lakeshore with agricultural influences, and the relative
extent and intensity of human influences of all types together). For each aggregated ecoregion,
a threshold value representing least disturbed conditions was established as a "pass/fail"
criterion for each parameter (Table 3-1). Thresholds were values that would be very unlikely to
be exceeded in least disturbed lakes within each region and varied by lake type to account for
regional variations in water chemistry and littoral-riparian human activities (Herlihy et al. 2013).
A lake was considered least disturbed if it passed the screening test for all parameters, and we
identified 214 least disturbed lakes from NLA 2012 and 168 from NLA 2007. We used the 2012
survey data for the 44 lakes from NLA 2007 that were again sampled in NLA 2012, and still
passed the reference screening, so 124 NLA 2007 lakes remained in the reference set (Table
5-1). Lakes that were not classified as least disturbed were provisionally considered
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intermediate in disturbance. The intermediate disturbance lakes were then screened with a set
of high-disturbance thresholds applied to the same variables (Table 3-2) Lakes that exceeded
one or more of the high disturbance thresholds were considered most disturbed. To avoid
circularity in defining physical habitat alteration, we did not use any of the physical habitat
cover complexity indices or their subcomponent metrics in defining lake disturbance classes.
Our screening process identified 382 least disturbed, 1309 intermediate, and 519 most
disturbed lake visits. Of the 338 least disturbed lakes that did not overlap survey years, 190
were in the WMT, NAP, and UMW aggregated ecoregions (Table 5-1). Even with relaxed
disturbance screening criteria, it was more difficult to find least disturbed lakes in some other
ecoregions. Respectively, only 11, 20, and 23 least disturbed lakes were identified in the NPL,
XER, and TPL ecoregions. To increase the useable sample size for estimating expected lake
condition, we grouped least disturbed lakes from the NPL, SPL, TPL into the Central Plains
(CENPL), and the WMT and XER into the West (for some models). Because of insufficient
numbers of least disturbed lakes relative to the large amount of lake variability within
ecoregions, we needed all available reference lakes for modeling expected conditions, so were
unable to use totally independent subsets of lakes for developing and validating those models.
5.3.4 Calculation of lake physical habitat metrics
5.3.4.1 Names of habitat metrics
Our variable names are those from the publicly-available NLA 2007,2012 and 2017 datasets
released by the EPA on the NARS Data webpage. The first several letters in the NLA variable
names denote the category and type of metric. The initial letters "hi..." identify human
influence metrics. The initial letters "hifp..." specify human influence frequency of presence
metrics and "hii..." specify indices of aggregated or summed human influences. Riparian
vegetation mean presence metrics begin with "rvfp ..." and mean riparian vegetation cover
metrics begin with "rvfc...", whereas "rvi..." denotes riparian vegetation cover sums (e.g., two
types of woody cover). The initial letters "fc..." and "am..." indicate, respectively, fish cover and
aquatic macrophyte metrics. These letters followed by "...fp...", "..fc...", or "..i.." indicate,
respectively mean frequency of presence among stations, mean areal cover, and indices
created by summing various metrics. Littoral bottom and exposed shoreline substrate metrics,
respectively, are identified by "bs..." and "ss...". The summary habitat indices described by
Kaufmann et al. (2014c), and used to define habitat condition in the NLA (RVegQ, LitCvrQ, and
LitRipCvQ) all end in the upper case Q, and the NLA summary human disturbance index is
RDis_IX (Riparian Disturbance Intensity and eXtent). Kaufmann et al. (2014a) describe in detail
the definitions and calculation of NLA physical habitat metrics and quantify their precision.
Many of the physical habitat metrics for NLA 2012 are additionally identified by the suffixes
_rip, _lit, and _DD (e.g., rviWoody_rip, rviWoody_DD, fciNaturalJit, fciNatural_DD),
designating that the habitat observations or measurements were from, respectively, the set of
riparian, littoral, or drawdown plots (Figure 5-1).
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5.3.4.2 Drawdown Zone Apportioning to match NLA 2007 Riparian and Human
Disturbance metrics:
NLA 2012 retained the measures of "bathtub ring" height and horizontal extent exactly as done
in NLA 2007 to quantify lake drawdown and seasonal lake level fluctuations. However, the near-
shore plot designs of the two surveys differ. In NLA 2007, the 15m x 15m riparian plots abutted
the shoreline. Consequently, exposed littoral bottom may comprise 0 to 100% of NLA 2007
plots, depending upon the extent of drawdown. Near-shore habitat was accurately depicted in
the NLA 2007 data, but because cover and disturbances were not separately assessed in the
drawdown zone, there was no accurate way to separately assess changes in habitat condition
attributable to drawdown (vs. riparian vegetation removal, for example). The NLA 2012 field
methods have separate measures of vegetation and human disturbances for the riparian and
drawdown zone plots, and separate fish cover estimates in littoral and drawdown zone plots.
These field plot changes improve the separation of lake level changes and drawdown from
other stressors in a diagnosis of likely causes of poor nearshore habitat condition in NLA 2012.
We used cover and human disturbance tally data from the riparian and drawdown plots to
calculate cover estimates or disturbance tallies simulating the set of ten 15m x 15m near-shore
plots abutting the shoreline, as had been used in the NLA 2007 field methods. We calculated
RCsyn, as a synthetic estimate of cover in the 15m band around the shoreline by summing the
areal covers in the drawdown and riparian plots, after weighting each by the proportion of the
15m band that was, respectively, within the drawdown zone or not within the drawdown zone:
RCsyn = (Rpdraw X RCdraw) "t" (Rprip X RCrip) (Eq 1)
where:
Rcsyn = Calculated cover in 15 x 15 m shoreline PHab plot, synthesizing metric values equivalent
to those used in NLA 2007, which represent the riparian condition in the 15m near-
shore band adjacent to the wetted edge of the lake.
Rpdraw and Rprip are the proportions of the 15x15m shoreline PHab plot that are, respectively,
occupied by the drawdown zone and the riparian zone above the high water mark.
[NOTE for NLA-2017 ONLY: There were a large number of missing measurements of
horizontal and vertical drawdown in the 2017 survey. The field protocol directs field
crews to NOT establish a drawdown plot when horizontal drawdown is <=lm. For 2017
we assumed drawdown was
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Rpdraw = (Horizontal Distance to high water)/(15m) = {bfxHorizDist/15m), and Rpdraw= 1.0 if
bfxHorizDist> 15 m.
RpriP= (1 - Rpdraw) by definition because RpriP+ Rpdraw= 1.0
Redraw and Rcnp are, respectively, the areal cover of vegetation in the drawdown and riparian
zones; Rcnp could be single cover type (e.g., canopy layer, or barren ground), or could
be a sum of cover types (e.g., sum of woody cover in 3 layers).
Calculated Rcsynfor a hypothetical lake with a mean horizontal drawdown of 10m (est. by
bfxHorizDist), and 100% canopy cover above the high water mark, but 0% cover in the
drawdown zone is as follows:
Rpdraw = 10/15 = 0.67
Rpnp =(1.0-0.67) = 0.33
Drawdown Canopy cover: RCdraw = 0%
Riparian Canopy cover: RcriP = 100%
RCsyn = (0.67 x 0%) + (0.33 x 100%) = 33%
The loss or gain in near-shore riparian habitat cover resulting from lake drawdown or natural
lake level declines can be estimated by the difference in cover between the riparian cover
above the high water mark (RcriP) and that within 15 m of the lakeshore (Rcsyn)-
We conducted a volunteer Drawdown Pilot Survey in 2011 to determine whether modification
of the NLA 2007 field protocols could be made without jeopardizing our ability to track changes
or trends in riparian habitat overtime (Anne Rogers 2012 NALMS; Kaufmann et al. Jan 9, 2012
webinar presentation to NLA steering committee and states). NLA 2007 and NLA 2012 field
protocols were applied simultaneously at 210 stations on 21 lakes spread over a range of
drawdown conditions in the states of Texas, Wisconsin, Washington, Oregon, Wyoming, North
Dakota, and Colorado. Kaufmann et al. (2012 webinar) demonstrated that 2007 metric values
for lakeshore vegetation and human disturbances were calculated accurately from the new
(2012) protocol, preserving ability to track changes/trends. The regressions predicting the
measured values of key physical habitat metric values from the NLA 2007 protocol from values
calculated by Eq 1 were virtually 1:1 lines with intercepts very close to 0.0, slopes very close to
1.0, and R2 between 0.87 and 0.94. The drawdown pilot analysis also showed that there was
virtually no difference in whole-lake metric values obtained by applying Eq 1 at each station,
versus applying it once per lake based on values of drawdown extent and cover averaged over
the 10 riparian and drawdown plots on each lake. The drawdown pilot results also
demonstrated that adding separate determinations of habitat cover elements in the drawdown
zone was logistically feasible and resulted in very minor increases in field time.
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5.3.4.3 Drawdown zone apportioning to estimate littoral habitat changes due to
drawdown:
We used a calculation similar to Eq 1 to simulate the amount of littoral cover that would be
present if, hypothetical^, the amount of lake drawdown was zero:
LCsim = (Lpdraw X l-Cdraw) + (LpiitX LClit) (Eq 2)
where:
LcSim = Calculated littoral cover simulating the amount of real or potential cover in a 10 x 15 m
littoral plot abutting the high-water mark, ie., simulating littoral cover that might be
present if there were no drawdown.
Lpdraw and Lpnt are the estimated proportions of a hypothetical 10m x 15m littoral PHab plot
abutting the highwater mark that are, respectively, occupied by the drawdown zone
(dry) and the littoral zone (wet). [NOTE for NLA-2017 ONLY: There were a large number
of missing measurements of horizontal and vertical drawdown in the 2017 survey. The
field protocol directs field crews to NOT establish a drawdown plot when horizontal
drawdown is <=lm. For 2017 we assumed drawdown was 10 m.
Lpnt= (1 - Lpdraw) by definition because LpriP+ Lpdraw= 1.0
Lent and LCdraw are, respectively, the areal cover of fish habitat elements in the littoral plot, and
exposed (dry) in the drawdown zone, Lc could be single cover type (e.g,,fcfcSnags) or
could be a sum of cover types (e.g., sum of non-anthropogenic cover types: fcfcNatural).
Calculated Lcs-,m for a hypothetical lake with a mean horizontal drawdown of 10m and 100%
Snag cover in the drawdown zone (dry and exposed), but 0% Snag cover in the littoral
(wet) zone is as follows:
Lpdraw= 10/10 = 1.00
Lpnt = (1.00-1.00) = 0
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Drawdown Snag cover: LCdraw= 100%
Littoral Snag cover: Lcnt= 0%
LCsim = (1.00 x 100%) + (0 x 0%) = 100%
The loss or gain in littoral habitat cover resulting from lake drawdown or natural lake level
declines can be estimated as the difference between the littoral cover simulated for zero
drawdown conditions (LcSim) the observed cover actually existing in the littoral at the time of
sampling {Lent).
5.3.4.4 Use of Variable suffixes in this document:
Riparian cover or human disturbance metrics calculated by Eq 1 are synthetic values that match
the 2007 metrics, and are designated by the suffixes_sy/7 (e.g., rviWoody_syn and hiiAII_syn) in
the EPA database. For simplicity, we will drop the suffixes on riparian vegetation and human
disturbance metrics in the remainder of this document, and it is understood that we are using
the synthesized variables when no suffix is present (*_syn), and NOT the drawdown zone
(*_DD), or riparian plot (*_r/p) versions of those variables.
Littoral cover metrics designated with the suffix_//t are based on field observations that are
conceptually and procedurally identical to those used in NLA 2007. For simplicity, we will drop
the suffixes on littoral cover metrics in the remainder of this documnet. and it is understood
that we are using the innudated littoral plot version of those variables when no suffix is
present f* lit), and NOT the drawdown zone (*_DD) or zero-drawdown simulated values
(*_s/m) versions of those variables. Littoral cover metrics calculated using Eq 2 simulate littoral
cover that would be present in the near-shore littoral area if the amount of drawdown were
zero, and are designated by the suffix_s/m (eg.,fciNatural_sim).
5.3.4.5 Near-shore disturbance metrics
We calculated extent of shoreline disturbance around the lakeshore (hifpAnyCirca) as the
proportion of stations at which crews recorded the presence of at least one of the 12
anthropogenic disturbance types as described by Kaufmann et al. (2014a). We calculated the
disturbance intensity metric hiiAII as the sum of the 12 separate proximity-weighted means for
all shoreline disturbance types observed at the 10 shoreline stations (Kaufmann et al. 2014a).
We also calculated subsets of total disturbance intensity by summing metrics for defined
groups of disturbance types. For example, hiiAg sums the proximity-weighted presence metrics
for row crop, orchard, and pasture; hiiNonAg sums the proximity-weighted presence metrics for
the remaining 9 non-agricultural disturbance metrics: 1) buildings, 2) commercial
developments, 3) parks or human-made beaches, 4) docks or boats, 5) seawalls, dikes, or
revetments, 6) trash or landfill, 7) roads or railroads, 8) power lines, and 9) lawns.
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5.3.4.6 Riparian vegetation metrics
Field data consisted of visual areal cover % class assignments of the vegetation type and areal
cover for each of 3 layers: canopy (>5 m high), mid-layer (0.5-5 m high), and ground cover (<0.5
m high). Crews estimated large (diameter at breast height [DBH] > 0.3 m) and small (DBH < 0.3
m) diameter tree cover separately in the canopy and mid-layer, distinguished woody from
herbaceous vegetation in the mid-layer and ground cover, and distinguished barren ground
from vegetation inundated by water in the ground layer. To characterize riparian vegetation in
the near-shore zone of the lake, we converted field cover class observations to mean cover
estimates for all the types and combinations of vegetation data (Kaufmann et al. 2014a). We
assigned cover class arithmetic midpoint values to each plot's cover-class observations (i.e.,
absent = 0%, sparse (>0-10%) = 5%, moderate (>10-40%) = 25%, heavy (>40-75%) = 57.5%, and
very heavy (>75-100%) = 87.5%), and then calculated lakeshore vegetation cover as the average
of those cover values across all 10 plots. Metrics for combined cover types (e.g., sum of woody
vegetation in 3 layers) were calculated by summing means for the single-types (see Kaufmann
et al. 1999, 2014a). Metrics describing the proportion of each lakeshore with presence (rather
than cover) of particular features were calculated as the mean of presence (0 or 1) over the 10
riparian plots.
5.3.4.7 Littoral cover and aquatic macrophyte metrics
The NLA survey crews made observations of the areal cover attributable to 8 littoral cover types
within each of the 10 littoral plots: rock ledges, boulders, brush, inundated live trees, snags,
overhanging vegetation, aquatic macrophytes, and human structures. Additionally, field crews
made separate visual estimates of areal cover for emergent, floating, and submerged aquatic
macrophytes within each of the 10 littoral plots. They used the same % cover classes for these
observations as used for riparian vegetation. Metrics describing the mean cover (and mean
presence) of littoral physical habitat features and aquatic macrophytes were calculated from
these cover class observations as described above for riparian vegetation. Metrics for combined
cover types (e.g. sum of natural types fish cover, floating and emergent aquatic macrophyte
cover) were calculated by summing means for single types.
5.3.4.8 Littoral and shoreline substrate metrics
NLA field crews visually estimated the percent areal cover of 8 substrate types (bedrock,
boulder, cobble, gravel, sand, silt/clay/muck, woody debris, and organic detritus) at each of the
10 near-shore stations (Figure 5-1). These estimates were made separately for the 1 m
shoreline band above the lake margin and for the lake bottom within the littoral plot. In cases
where the bottom substrate could not be observed directly, crews viewed the bottom through
a viewing tube, felt the substrate with a 3 m PVC sounding tube, or observed sediments
adhering to the boat anchor as it was retrieved from the bottom. Cover classes were the same
as for riparian vegetation. We calculated metrics describing the lake-wide mean cover of near-
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shore littoral and shoreline substrate in each size category by averaging the cover estimates at
each station, based on the cover class midpoint approach described above.
We adapted the approach of Faustini and Kaufmann (2007) and Kaufmann et al. (2009) for
estimating geometric mean and variance of substrate diameters from systematic pebble-
counts. In this approach (Kaufmann et al. 2014a), we assigned the geometric mean between
the upper and lower diameter bound of each size class for each cover observation before
calculating the cover-weighted mean size index. We calculated the geometric mean diameters
(Dgm) of littoral and shoreline substrate (bsxLdia and ssxLdia) as follows:
Dgm=Antilog{Sum/{P/{[logio(D/u)+logio(D// )]/2}}}, (Eq. 3)
where:
P,=areal cover proportion for diameter class /';
D/u=diameter (mm) at upper limit of diameter class /';
Dn =diameter (mm) at lower limit of diameter class /';
Sum, =summation across diameter classes; and
Nominal size class midpoint diameters of 5660 and 0.0077 mm were set, respectively, for the
largest (bedrock and hardpan) and smallest (silt, clay, and muck) diameter classes.
Our calculations are identical to those of Faustini and Kaufmann (2007), except that here the
percent cover estimates used to weight diameters were the mean values of 10 visual cover
estimates rather than areal streambed cover determinations derived from the pebble-count
percentages for individual particles in each diameter class.
5.3.4.9 Littoral depth, Lake level fluctuations, bank and water surface characteristics
Field crews measured littoral depth, estimated water level fluctuations and bank heights, and,
and observed water surface and bottom sediment color and odor at each of the 10 nearshore
stations (Figure 5-1). SONAR, sounding lines, or sounding tubes were used to measure lake
depth 10 m offshore. NLA field crews used hand-held levels, survey rods, and laser rangefinders
(rather than unaided visual estimates) to measure vertical and lateral (horizontal) lake level
fluctuation. Field indications of short to medium term fluctuation, drawdown and/or declines in
lake levels were based on measurement of the vertical height and horizontal extent of exposed
lake bottom ("Bathtub Ring") field evidence.
Crews recorded the presence of surface films or scums, algal mats, oil slicks, and sediment color
and odor. They visually estimated the bank angle in the 1 m-wide shoreline band and the
vertical and lateral range in lake level fluctuations, based on high and low water marks. We
calculated whole lake metrics for mean littoral depth and water level fluctuations as arithmetic
averages (sixDepth, bfxVertHeight and bfxHorizDist) and standard deviations of the measured
values at the 10 stations. For bank angle classes and qualitative observations of water surface
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condition and sediment color and odor, we calculated the proportion of stations having
observations in each class.
5.3.5 Calculation of summary physical habitat condition indices
We calculated 4 multimetric indices of physical habitat condition and an index of lake
drawdown:
RDis_IX: Lakeshore Anthropogenic Disturbance Index (Intensity and Extent),
RVegQ: Riparian Vegetation Cover Complexity Index,
LitCvrQ: Littoral Cover Complexity Index,
LitRipCvQ: Littoral-Riparian Habitat Complexity Index, and
Drawdown Index: based on bfxVertHeight and bfxHorizDist
5.3.5.1 Lakeshore Anthropogenic Disturbance Index (RDis_IX)
This index was calculated as:
RDis_IX = (Disturbance Intensity + Disturbance Extent)/2; (Eq 4)
where :
disturbance intensity was represented by separate sums of the mean proximity-weighted tallies
of near-shore agricultural and non agricultural disturbance types and extent was expressed as
the proportion of the shore with presence of any type of disturbance.
1
1-
RDis IX =
[l + hiiNonAg + (5 x hiiAg)]
+ hifpAnyCirca
(Eq 5)
2
where:
hiiNonAg = Proximity-weighted mean disturbance tally (mean among stations) of up to 9
types of non-agricultural activities.
hiiAg = Proximity-weighted mean tally of up to 3 types of agriculture-related activities
(mean among stations).
hifpAnyCirca = Proportion of the 10 shoreline stations with at least 1 of the 12 types of
human activities present within their 10 x 15 m littoral plots, drawdown plots, or within
15m of the lake shore in their 15 x 15 m riparian plots.
Field procedures classified only 3 types of agricultural disturbances, versus 9 types of non-
agricultural disturbances, limiting the potential ranges to 0-3 for hiiAg and 0-9 for hiiNonAg. In
the combined NLA 2007 and 2012 surveys, the observed ranges of these variables also differed:
hiiAg ranged from 0 to 1.55, whereas hiiNonAg had an observed range almost 5 times as great
(0 to 7.125). To avoid under-representing agricultural disturbances and over-representing non-
agricultural disturbances in the index, we weighted the disturbance intensity tallies for
agricultural land use by a factor of 5 in Equation 2. This weighting factor (ratio of observed
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ranges in non-agricultural to agricultural disturbance types) effectively scales agricultural land-
uses equal in disturbance potential to those for non-agricultural land uses. We scaled the final
index from 0 to 1, where 0 indicates absence of any anthropogenic disturbances and 1 is the
theoretical maximum approached as a limit at extremely high disturbance. We applied a single
formulation of the disturbance index RDis_IX throughout the NLA survey in the U.S.
5.3.5.2 Riparian vegetation cover complexity index (RVegQ)
This index is based on visual estimates of vegetation cover and structure in three vegetation
layers at the 10 near-shore riparian plots along the lake shore. The cover metrics were
calculated for the variable-width drawdown zone plots (metrics with suffix "_DD") and the 15m
x 15m riparian plots (with suffix "_rip"). For the NLA 2012 report, we used areal cover
information from both types of plots along with drawdown horizontal extent information to
calculate RVegQ estimates matching those for the previous report, which are for the near-
shore zone extending from the lake water's edge 15m outward (see Eq. 1). Because the
potential vegetation cover differs among regions, we calculated three variants of the Riparian
Vegetation Cover-Complexity Index (RVegQ^2, RVegQ^7, or RVegQ_8) for application to
different aggregated ecoregions (Table 5-2). The region-specific formulations reduce the
among-region variation in index values in least disturbed lakes and reduce ambiguity in their
response to anthropogenic disturbances. If component metrics had potential maximum values
>1, their ranges were scaled to range from 0 to 1 by dividing by their respective maximum
values based on the NLA 2007 data (see Table 3 in Kaufmann et al. 2014a). Each variant of the
final index was calculated as the mean of its component metric values. Index values range from
0 (indicating no vegetative cover at any station) to 1 (40 to 100 % cover in multiple layers at all
stations).
^ rvi Woody
2.5
RVegQ _ 2 =
RVegQ_7 =
RVegQ _ 8 =
+ rvfc (in din un dated
rviLowWood
1.75
+ rvfcGndlnundated
rviWoody
2.5
+ rvfpCanBig + rvfcGndlnundated + ssiNATBedBld
(Eq 6)
(Eq 7)
(Eq 8)
where:
rviWoody = Sum of the mean areal cover of woody vegetation in 3 layers: canopy (large and
small diameter trees), understory, and ground layers (rvfcCanBig + rvfcCanSmall +
rvfcUndWoody + rvfcGndWoody).
rviLowWood = Sum of mean areal cover of woody vegetation in the understory and ground
cover layers (rvfcUndWoody + rvfcGndWoody).
rvfcGndlnundated = Mean areal cover of inundated terrestrial or wetland vegetation in the
ground cover layer.
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rvfpCanBig = Proportion of stations with large diameter (>0.3 m dbh) trees present.
ssiNATBedBId = Sum of mean areal cover of naturally-occurring bedrock and boulders
(ssfcBedrock + sfcBoulders), and where the value of ssiNATBedBId was set to 0 in lakes
that have a substantial amount of human-built seawalls and revetments (i.e., hipwWalls
>0.10).
We used RVegQ_2 for mesic ecoregions with maximum elevations <2,000 m (NAP, SAP, UMW,
CPL) where tree vegetation can be expected in relatively undisturbed locations (Table 5-2).
RVegQ_2 sums the woody cover in three lakeside vegetation layers (rviWoody) and includes
inundated groundcover vegetation (rvfcGndlnundated) as a positive characteristic.
We used RVegQ_7 for Central Plains ecoregions (NPL, SPL and TPL). Whereas perennial woody
groundcover and shrubs can be expected on undisturbed lake shorelines throughout the
Central Plains (West and Ruark 2004), the presence or absence of large trees (>5m high) along
lake margins in this region has ambiguous meaning without floristic information (Johnson 2002,
Barker and Whitman 1988, Huddle et al. 2011). RVegQ_7 accommodates lack of tree canopy in
least disturbed lakes by summing only the lower 2 layers of woody vegetation (rviLowWood)
and includes inundated ground cover vegetation as a positive characteristic.
We used RVegQ_8 for the West (WMT, XER), where climate ranges from wet to arid, and where
lakeshores may have the potential to grow large diameter riparian trees but may lack vegetated
lake shorelines at high elevations, or where rock precludes vegetation (Table 5-2). RVegQ_8
sums the woody cover in 3 lakeside vegetation layers and includes inundated groundcover
vegetation as a positive characteristic; it also includes the proportional presence of large
diameter trees around the lakeshore as a positive characteristic. RVegQ_8 includes natural rock
as an undisturbed riparian cover type to avoid penalizing relatively undisturbed lakes in arid
areas or at high elevations above timberline. For lakes where there is a substantial extent or
abundance of constructed seawalls, dikes, or revetments along the shoreline, the substrate
metric was set at 0.
5.3.5.3 Littoral cover complexity index (LitCvrQ)
This index was based on the station-averages for visual estimates of the areal cover of 10 types
of littoral features, including aquatic macrophytes but excluding human structures, within each
of the 10 littoral plots (see Kaufmann et al. 2014a). Note that littoral metrics used to calculate
LitCvrQ are those with the suffix "_Hf, which match exactly the NLA 2007 littoral cover metrics
having no suffix. We calculated 3 variants, for application in different ecoregions (Table 5-2).
Each variant of the index was calculated as the mean of its component metric scores, so index
values range from 0 (no cover present at any station) to 1 (very heavy cover at all 10 stations).
Component metrics with potential maximum values >1 were scaled from 0-1 by dividing by
their respective maximum values in the NLA 2007 dataset.
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fciNatural+
LitCvrQ _b =
LitCvrQ _c =
LitCvrQ d =
fcfcSnag
0.2875 ,
fciNatural +
fcfcSnag (amfcFltEmg
v
0.2875
1 515
r SomeNatCvr
v is .
fcfcSnag (amfcFltEmg
0.2875
1 515
(Eq 9)
(Eq 10)
(Eq 11)
where:
fciNatural = summed areal cover of non-anthropogenic fish cover elements (fcfcBoulders +
fcfcBrush + fcfcLedges + fcfcLivetrees + fcfcOverhang + fcfcSnag + fcfcAquatic).
SomeNatCvr = summed cover of natural fish cover elements excluding snags and aquatic
macrophytes (fcfcBoulders + fcfcBrush + fcfcLedges +fcfcLivetrees + fcfcOverhang).
amfcFltEmg = summed cover of emergent plus floating aquatic macrophytes (amfcEmergent +
amfcFloating).
fcfcAquatic = total cover of aquatic macrophytes of any type.
All three variants of LitCvrQ include an expression of the summed cover of naturally occurring
fish or macroinvertebrate cover elements. Snag cover is recognized as a particularly important
element of littoral habitat complexity (Francis and Schindler 2006, Christensen et al. 1996,
Miranda et al. 2010). Therefore, we included snags as a separate contributing cover component
in all three variants of the index, and divided cover metrics by their maximum values in the NLA
2007 data to make the weightings of snag cover equal to those of the other two littoral cover
sums. For LitCvrQ^c and LitCvrQ_d, we increased the emphasis on emergent and floating-leaf
aquatic macrophytes relative to other littoral components in response to their reported
importance as cover and their sensitivity to human disturbances in many lake types and regions
(Radomski and Geoman 2001, Jennings et al. 2003, Merrell et al. 2009, Beck et al. 2013).
We used LitCvrQ^b for lakes in the CPL, which includes many generally shallow, warm, low
conductivity lakes. We used LitCvrQ_c for lakes in the SAP, which are all reservoirs, where
disturbed sites commonly have substantial erosion of clay-rich upland soils, large water level
fluctuations, and bare-soil shorelines. These conditions generate abiotic turbidity that
suppresses submerged macrophytes, thereby diminishing the association of abundant
submerged aquatic macrophytes with anthropogenic nutrient inputs that is typically seen in
other regions. LitCvrQ^c emphasizes floating and emergent aquatic macrophytes in addition to
snags, but still includes submerged aquatic macrophytes along with other aquatic macrophytes
and cover types in fciNatural. LitCvrQ_d excludes submerged aquatic macrophytes, and we
used it in the remaining ecoregions (NAP, TPL, NPL, SPL, WMT, and XER), where submerged
aquatic macrophytes provide valuable cover, but high submerged cover is frequently associated
with anthropogenic eutrophication (Hatzenbeler et al. 2004, Merrell et al. 2009).
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5.3.5.4 Littoral-riparian habitat complexity index (LitRipCvrQ)
We averaged the lake values of the littoral cover complexity and riparian vegetation cover
complexity indices to calculate the littoral-riparian habitat complexity index LitRipCvrQ:
T.„. _ _ (RVegQ n + LitCvrQ x)
LitRipCvrQ = =— ; (Eq 12)
where:
RVegQ_n = variant of the riparian vegetation cover complexity index (n=2, 7 or 8, depending on
ecoregion, Table 5-2.
LitCvrQ_x = variant of littoral cover-complexity index (x = b, c, or d, depending on ecoregion,
Table 5-2.
5.3.5.5 Lake level drawdown index (combined use of bfxVertHeight and bfxHorizDist)
We used the mean lake values estimating Lake Level Vertical Fluctuation (bfxVertHeight) in
combination with Lake Level Horizontal Fluctuation (bfxHorizDist) to characterize lake
drawdown and natural lake level declines. These metrics are, respectively, the height (meters)
measured from the present lake level to high water, and the horizontal (lateral) distance in
meters from the lake shore to the high water mark in meters. NLA field crews made these
determinations based on the extent and location of vegetation intolerant to frequent or
prolonged inundation, location of flotsom deposits ("trash racks"), evidence of wave action,
and exposed lake bottom. The lake bottom exposure measured by these methods characterizes
seasonal lake level declines and fluctuations on timescales shorter than that required for
disintegration of flotsom at the high water mark, or encroachment of perennial terrestrial
vegetation onto the exposed lake bottom area. In most regions, these measurements should be
adequate to document trends in lake level declines attributable to climate change, water
withdrawals, and reservoir management over a decadal timescale. However, more rigorous
tracking of such trends over longer timescales would require that field crews measure lake
levels in relation to established permanent (monumented) reference elevations and/or staff
gauges at sample lakes.
5.3.6 Deriving expected index values under least disturbed conditions
We based expectations for bfxVertHeight arid bfxHorizDist on "Null Models": the expected
value and its dispersion are represented by the central tendency and distribution of these
variables in regional sets of least disturbed reference sites. In the CENPL and WEST,
expectations were set separately for natural lakes versus human-made reservoirs.
We used lake-specific predictive regression models to estimate physical habitat expectations
for RVegQ, LitCvrQ, and LitRipCvrQ under least disturbed condition (Table 5-3). We compared
the performance of these regression models with null models (Table 5-4), for which
expectations were simply the mean of logio-transformed physical habitat index scores among
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least disturbed lakes from each ecoregion. Our motivation for using lake-specific models of
expected ("E") condition was to reduce the variance in physical habitat condition indices (in this
case O/E values of RVegQ, LitCvrQ, and LitRipCvrQ) among least disturbed reference lakes. Air
temperature, precipitation, soils and lithology can vary greatly across ecoregions, resulting in
corresponding variations in potential natural vegetation among least disturbed lakes. In turn,
that variation results in differences in the amount and complexity of littoral cover, especially for
those elements derived from riparian vegetation. We derived lake-specific expected values by
modeling the influence of important non-anthropogenic environmental factors in relatively
undisturbed lakes, an approach analogous to that used to predict least disturbed conditions for
multimetric fish assemblage indices (Esselman et al. 2013, Pont et al. 2006, 2009).
For calculating lake-specific expected (E) values of RVegQ, LitCvrQ, and LitRipCvrQ under least
disturbed condition, we conducted the multiple linear regression (MLR) modeling in 7
aggregated ecoregions (Table 5-3 and Appendix A). These models were based on least
disturbed lakes from the combined NLA 2007 and 2012 surveys within each region (Table 5-1).
The lake habitat index MLRs employed one to four predictors from among the following:
Latitude, Longitude, Elevation, ElevXLatitude, ElevXLongitude, Lake surface area, Lake origin
(human-made reservoir or natural lake), near-shore anthropogenic disturbance of all types
(RDis_IX), and near-shore anthropogenic agricultural disturbance (hiiAg). Latitude, longitude,
elevation, and ecoregion are surrogates for temperature, precipitation, soil, and other
characteristics that influence potential natural vegetation and littoral cover. Field
measurements of bfxVertHeight and bfxHorizDist were good predictors of riparian and littoral
cover in most of the regions. However, we chose not to use these indicators of level fluctuation
and drawdown to predict expected condition because their use would confound interpretations
and obscure the effects of drawdown on habitat condition. We also did not use lake depth
measurements (like maximum depth or littoral mean depth), because of their association with
lake level change. Similarly, survey year was a good predictor of lake physical habitat metrics in
regions where there were marked differences in the amount of lake drawdown between
surveys. We chose not to use survey year as a predictor of expected condition because it would
confound analysis of temporal trends and change between surveys.
Ideally, calculations of expected cover and complexity would be based only on minimally-
disturbed lakes. However, the least disturbed lakes in most regions include sometimes
substantial disturbances, necessitating inclusion of near-shore disturbance predictors in our
models if they were associated with variance in the habitat indices. The use of RDis_IX or hiiAg
as predictors was supported by the data for all three habitat indicators in the NPL, CPL and
CENPL, and the littoral cover indicator in the SAP (Table 5-3). For predicting expected LitCvrQ
and LitRipCvrQ in the NAP, we had to combine least disturbed with moderately disturbed lakes
and reservoirs (RT_NLA12_2015 = R or S) to span lake size and elevation gradients affecting
riparian vegetation and littoral cover in that region. The weak association of human disturbance
with habitat indices would not have warranted including RDis_IX as a predictor within NAP least
disturbed sites alone (RT_NLA12_2015=R). However, the human disturbance gradient
introduced by including moderately disturbed NAP lakes (RT_NLA12_2015=S), and the effect of
that disturbance on littoral habitat in the NAP made it necessary to include RDis_IX as a
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predictor. Inclusion of RDis_IX or hiiAg as predictors of expected lake habitat index values was
not supported by the data for lakes and reservoirs in the UMW, WMT, and XER. As in most of
the other regions, lake level fluctuation indicators were good predictors of riparian and littoral
cover in the UMW and WEST, but were not used as predictors for reasons we stated in the
previous paragraph.
For regions where RDis_IX or hiiAg were used in modeling expected habitat condition, we set
the value of these variables in the predictive MLR equation to the minimum value observed in
the region before calculating expected values of RVegQ, LitCvrQ, and LitRipCvrQ. In all regions
and subregions there were sites with RDisJX and hiiAg values of 0 (See Appendix A). Setting the
reference expected lake habitat index values slightly higher in this way results in the central
tendency for reference site 0/E to be less than 1.0.
5.3.7 Condition criteria for nearshore lake physical habitat
For the lakeshore anthropogenic disturbance index RDisJX, we used uniform criteria for all
lakes. For RVegQ, LitCvrQ, and LitRipCvQ we set condition criteria based on the distribution of
O/E values of these indices observed in least disturbed lakes. For bfxVertHeight and
bfxHorizDist, we set condition criteria based on the distribution of the metric values themselves
in least disturbed lakes (Null model).
5.3.7.1 Condition Criteria for Lakeshore Anthropogenic Disturbance Intensity and Extent
Because RDisJX is a direct measure of human activities, we based criteria for high, medium,
and low levels of disturbance on judgment:
Good (Low Disturbance): RDisJX <0.20
Fair (Medium Disturbance): RDis_IX >0.20 but < 0.75
Poor (High Disturbance): RDis_IX>0.75
Lakes with RDisJX <0.20 have very low levels of lake and near-lake disturbance, typically having
anthropogenic disturbance on <8% of their shorelines. Those with RDisJX >0.75 have very high
levels of disturbance, typically having human activities evident on 100% of their shorelines. For
perspective, <21% of the 2364 sample site visits in the combined NLA 2007 and 2012 surveys
had RDisJX <0.20, and <21% had RDisJX >0.75. Most of the reference sites in the WMT, UMW,
and NAP regions have RDisJX <0.20, most of those in SAP, SAP, XER, TPL, and CPL have RDisJX
<0.40, most NAP reference sites have RDisJX between 0.40 and 0.6, and no reference sites
have RDisJX >0.70 (Figure 5-3).
5.3.7.2 Condition Criteria for RVegQ, LitCvrQ, and LitRipCvQ
We calculated physical habitat index observed/expected (O/E) values of RVegQ^OE,
LitCvrQ^OE, and LitRipCvQ_OE for each sample lake by dividing the observed index value at
each lake by the lake-specific expected value derived from regressions in Table 5-3 and
Appendix A. The calculated O/E values of the habitat metrics for each lake express the degree
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of deviation of that lake from an estimate of its expected value under least disturbed
conditions. No model perfectly predicts expected indicator values (E-values) in lakes under least
disturbed conditions, and field measurements of indicator values ("0" values) include error and
temporal variation. Consequently, 0/E values of these indices among reference lakes have a
dispersion (variance) that decreases with the performance of predictive models (i.e., how
precisely does the model predict reference condition?), and with the precision of the habitat
indicator measurements (i.e., how well do the field methods measure observed condition?). We
set condition criteria for RVegQ, LitCvrQ, and LitRipCvQ with reference to the distributions of
these indices among least disturbed lakes within each of the 7 merged ecoregions Table 5-5.
The small number of lakes meeting our low-disturbance criteria in most regions precluded
obtaining reliable percentiles of RVegQ, LitCvrQ, and LitRipCvQ directly from the least disturbed
lake distributions. Consequently, for all regions, we used the central tendency and variance of
index 0/E values in least disturbed lakes values to model their distributions and to estimate
percentiles (Snedecor and Cochran 1980). The logio-transformed 0/E values in the least
disturbed lakes had symmetrical, approximately normal distributions. We calculated means and
standard deviations of logio-transformed 0/E values (Table 5-5, columns 3 and 4), and
estimated the 5th and 25th percentiles (Table 5-5, columns 7 and 8) based on the log-normal
approximation of the index distributions in least disturbed lakes within each ecoregion. Because
the means and SD's are all log values, a range of + 1SD would be calculated, for example, by
multiplying and dividing the geometric mean by the geometric SD (see Table 5-5 legend for
details, including handling of the log-transformation constant).
Lakes with 0/E values (MLR model) that are >25th percentile for least disturbed lakes within
their regions were considered to have habitat in good condition (i.e., similar to that in the
population of least disturbed lakes of the region). Similarly, lakes with index or 0/E values <5th
percentile of least disturbed lakes were considered to have poor habitat quality (i.e., they have
significantly lower cover and complexity than observed within the sub-population of least
disturbed lakes of the region). Those with index or 0/E values between the 5th and 25th
percentiles of least disturbed lakes were scored as fair condition.
We emphasize that our designations of good, fair and poor are relative to the least disturbed
sites available in each ecoregion. We define good condition as habitat quality not
distinguishable from the distribution of habitat in least disturbed sites; and poor condition as
habitat quality that is not likely to be found within the distribution of least disturbed sites of the
ecoregion. Our designations of poor condition do not indicate impaired water body status.
Conversely, our designations of good condition mean that habitat is similar to the least
disturbed sites available in a region, which does not mean pristine, only the best available,
which can be relatively disturbed in extensively and most disturbed regions.
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5.3.7.3 Condition Criteria for Lake Drawdown
We based our assessment of Lake Drawdown condition on null models of the expected amount
of drawdown in least disturbed lakes. Specifically, we examined the empirical distributions of
the metrics quantifying vertical and horizontal lake level fluctuations [bfxVertHeight and
bfxHorizDist) in least disturbed lakes within aggregated ecoregions, sometimes stratified by lake
origin (natural lakes versus human-made reservoirs). We used separate null models for the
NAP, SAP, UMW, and CPL regions. For the CENPL (TPL+SPL+NPL) and the West (WMT+XER), we
used separate null models for natural lakes versus human-made reservoirs. Vertical and
horizontal drawdown were considered small if they were <75th percentile of their respective
reference distributions; large if >95th percentile, and medium if in-between (Table 5-6). Overall
lake drawdown condition was considered small if both vertical and horizontal drawdown were
small; medium if one or both were medium (but not large); and large if vertical, horizontal or
both were large.
NOTE for NLA 2017 ONLY:
In several hundred NLA-2017 sample lakes, field crews did not measure horizontal or vertical
drawdown in cases where they did not establish drawdown zone cover plots. In these cases, we
assumed that missing horizontal drawdown values were
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5.4.2 Null model results for RVegQ, LitCvrQ, and LitRipCvQ:
Geometric means for RVegQ, LitCvrQ, and LitRipCvQ in least disturbed lakes differed among
regions (Table 5-4), but these unsealed null model values are not directly comparable because
the habitat index formulations differed among regions. The RVegQ, LitCvrQ, and LitRipCvQ null-
model logSD's and geometric SD's (Columns 4 and 6 of Table 5-4) were calculated from log-
transformed variables, and therefore are expressions of the proportional variance among least
disturbed lakes of each region. Whether scaled (divided by the mean) or not, they are directly
comparable as measures of model precision among regions with different geometric means, or
between null and MLR modeling approaches.
Comparing indicators, the precision in modeling least disturbed condition using null models was
generally better (smaller SDs) for LitRipCvQ than for RVegQ or LitCvrQ, and null models for
RVegQ were generally more precise than for LitCvrQ (Table 5-4, columns 4 and 6). The most
obvious differences, however, were among regions, and the differences were associated with
the level of disturbance in the reference sites. We ordered the seven NLA lake habitat modeling
ecoregions according to increasing reference site median RDisJX for examining variance in the
other lake habitat indicators (Figure 5-3). The regions with the greatest amount of disturbance
in their reference sites (the CENPL, including NPL, SPL, TPL, the CPL, and the XER) generally had
higher within-reference site variance all three lake habitat indices, with the exception of low
variance in all three indicators within reference sites of the relatively high-disturbance CPL
reference sites (Figure 5-4). The precision in modeling least disturbed condition using null
models was generally best in the UMW and NAP (i.e., lowest gSDs). The smaller the SD of index
values (or O/E values) among least disturbed lakes, the easier it is to confidently distinguish
disturbed lakes from least disturbed lakes. The null model SD's serve as an upper bound for the
variance of the indicators among regional reference sites, and are analogous to the RMSE's of
the regressions in Table 5-3. Removing the variance attributed to the predictors reduces the
unexplained variance among reference sites.
5.4.3 O/E model results for RVegQ, LitCvrQ, and LitRipCvQ:
The LogSD's of RVegQ^OE, LitCvrQ^OE, and LitRipCvQ_OE among reference sites (Table 5-5,
column 4) were consistently, and in some cases substantially, lower than those for null models
in their respective regions, as evidenced by comparing open circles and black dots plotted in
Figure 5-4. The CPL, CENPL, XER and WMT showed the largest reduction of reference site
variance compared with corresponding null models, denoting improvement in O/E model
performance over null models. As for the null models, however, O/E models in regions with
relatively disturbed reference sites had higher reference site variance (the expected condition
models were less precise). Again, with the exception of the CPL, regions with more disturbance
in their reference sites still had higher SD's than those in regions with less disturbance.
Conversely, the four regions with the lowest level of human disturbance in their reference sites
(WMT, UMW, NAP, and SAP) also had the lowest O/E model variance among their reference
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sites. These results reinforce the idea that human disturbances are likely responsible for a large
amount of the variance in lake physical habitat structure in reference sites within the disturbed
regions. Therefore, further effort to capture this variance by modeling only non-anthropogenic
("natural") controls would not likely be successful in reducing the variance in O/E values among
reference sites.
Except for regions where O/E models incorporated human disturbance variables (NAP, CPL,
CENPL and LitCvr_OE in SAP), the central tendency of reference site O/E values (Table 5-5,
column 6) was very close to 1 (0.98 to 1.01). This is to be expected. Where E-Models contained
human disturbance predictors, reference O/E values regained the variance modeled out when
observed values were divided by expected values determined with human disturbance
predictors (RDis_IXor hiiAg) set to regional minimum values. If human disturbances decrease
the observed value, the mean O/E will be <1. Accordingly, reference site mean O/E values for
MLR Models in the NAP, CPL, and CPL (and LitCvr_OE in SAP) ranged from 0.79 to 0.91. We
regressed the reference O/E values against the RDis_IX or hiiAg values to obtain y-intercepts for
expected O/E for the minimum disturbance observed in these regions. These are shown in the
Table 5-5 rows with "oe Yint" subscripted after their Ecoregion designation. For example the
NAPoEYint row is the result of this final adjustment on reference O/E results from the NAPMLRModei
row.
Anthropogenic disturbance among reference sites tends to increase the variance in O/E values
within regions, even after the minimum disturbance adjustment. There is a strong relationship
between the LogSDs of null and adjusted O/E models for lake habitat among reference lakes
and the regional level of near-shore anthropogenic disturbance in reference sites (Figure 5-4).
Our modeling improves these models, but it is likely that disturbances other than those
captured by RDis_IXcontribute to the uncertainty in predicting habitat characteristics in
minimally-disturbed lakes. These results reinforce the idea that human disturbances are likely
responsible for a large amount of the variance in lake physical habitat structure among least
disturbed reference sites in the disturbed regions. Therefore, further effort to capture this
variance by modeling only non-anthropogenic ("natural") controls would not likely be
successful in reducing the variance in O/E values among reference sites.
5.4.4 Null model results for lake drawdown and level fluctuations:
Least disturbed reference lakes and reservoirs in the NAP, SAP and UMW experienced less
drawdown and level fluctuation than those in the CPL, CENPL, and WEST; particularly in
comparison with marked drawdown observed in human-made reservoirs of the CENPL and
WEST (Table 5-6). Not surprisingly, least disturbed natural lakes in the CENPL and WEST also
experienced less drawdown and level fluctuation than their human-constructed counterparts.
As a result, the criteria for assessing substantial drawdown in lakes of the Appalachians and
UMW were much smaller than those for lakes (and particularly reservoirs) in the CENPL and
WEST.
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5.5 Precision of physical habitat indicators
In our synoptic survey context, o2iake is the signal of interest, and o2rep is noise variance; we
define their ratio as S/N. The methods we used to quantify precision, the precision of NLA lake
physical habitat metrics and key habitat condition indices, and the implications of varying
precision levels for monitoring and assessment, are comprehensively evaluated by Kaufmann et
al. (1999, 2014a). Here we summarize findings for key physical habitat indicators based on the
NLA 2012 survey data, which is a good representation of precision for NLA 2017, based on
Kaufmann et al. (2014a) and the NLA 2012 Technical Support Document (USEPA 2017b).
The key NLA physical habitat indices had moderate to high S/N (2.2 - 11.0) over the entire NLA
2012 survey (Table 5-7). Compared with the other composite indices, the human disturbance
index RDis_IX and horizontal drawdown index had the highest S/N (9.1-11), whereas the littoral
cover O/E index had the lowest S/N (2.2). The advantage of S/N as a precision measure is its
relevance to many types of statistical analysis and detecting differences in subpopulation
means (Zar 1999). High noise in habitat descriptions relative to the signal (i.e., low signal: noise
ratio, S/N) diminishes statistical power to detect differences among lakes or groups of lakes.
Imprecise data limit the ability to detect temporal trends (Larsen et al. 2001, 2004). Noise
variance also limits the maximum amount of variance that can be explained by models such as
multiple linear regression (Van Sickle et al. 2005, Kaufmann and Hughes 2006). By reducing the
ability to quantify associations between variables (Allen et al. 1999, Kaufmann et al. 1999),
imprecision compromises the usefulness of habitat data for discerning likely controls on biota
and diagnosing probable causes of impairment. The adverse effects of noise variance on these
types of analysis are negligible when S/N >10; becoming minor as S/N decreases to 6, increasing
to moderate as S/N decreases to 2, and finally becoming severely limiting as S/N approaches 0
(Paulsen et al. 1991, Kaufmann et al. 1999). At S/N=0, all the metric variance observed among
lakes in the survey can be attributed to measurement "noise". Based on these guidelines, the
effects of imprecision are minor for all the indicators except for the Littoral Cover index, for
which the effects are minor-to-moderate.
Kaufmann et al. (2014a) explain that the S/N ratio may not always be a good measure of the
potential of a given metric to discern ecologically important differences among sites. For
example, a metric may easily discriminate between sparse and abundant littoral cover for fish,
but S/N for the metric would be low in a region where littoral cover does not vary greatly
among lakes. In cases where the signal variance (o2iake) observed in a regional survey reflects a
large range of habitat alteration or a large range in natural habitat conditions, S/N would be a
good measure of the precision of a metric relative to what we want it to measure. However, in
random surveys or in relatively homogeneous regions, o2iake and consequently S/N, may be less
than would be calculated for a set of sites specifically chosen to span the full range of habitat
conditions occurring in a region. To evaluate the potential usefulness of metrics, Kaufmann et
al. (2014a) suggested that an alternate measure of relative precision, orep divided by its
potential or observed range (Rgpot or Rg0bs) offers additional insight. The minimum detectable
difference in means between 2 lakes (or between two times in one lake) is given by Dmin =
1.96arep(2n)1/2 = 2.77arep , using a 2-sided Z-test with a = 0.05 (Zar 1999). Thus, to detect any
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specified difference between 2 lakes in a metric relative to its potential or observed range [Rgpot
or Rg0bs, the standardized within-lake standard deviation, orep/Rg, cannot exceed (Dm/n/
Rg)/2.71. By the criteria in Kaufmann et al. (2014a - Table 2), the key NLA physical habitat
indices were precise or moderately precise, with orep/Rg0bs between 0.052 - 0.107 (Table 5-7).
Depending on the index, they have the potential to discern differences between single lakes (or
one lake at two different times) that are between l/3rd and l/8th the magnitude of the
observed ranges of these indices.
5.6 Physical habitat index responses to anthropogenic disturbance
In the U.S. as a whole, RVegQ_OE, LitCvrQ^OE, and LitRipCvQ_OE were significantly higher
(p<0.0001) in least disturbed lakes (RT_NLA12_2015=R) than in most-disturbed lakes
(RT_NLA12_2015=T) (Table 5-8, Figure 5-5). The differences were substantial for RVegQ_OE,
and LitRipCvQ_OE, and discrimination was good (no or nearly no overlap in interquartile
ranges). For LitCvrQ^OE, there was an overlap of approximately one-third of the interquartile
range. RDisJX was a major screening variable used to disqualify potential reference sites, so it
is not surprising that the entire range of RDisJX among reference sites had very little overlap
with that for most disturbed sites. Note that a site with very low RDisJX could be classified as
most-disturbed on the basis of many other variables, but the converse is not true because
reference sites must all have low RDisJX. Like RDisJX, both vertical and horizontal drawdown
were significantly lower (p<0.0001) in least disturbed lakes than in most-disturbed lakes (Table
5-8, Figure 5-5). Except for lake drawdown, contrasts were very similar for the NLA 2007 and
2012 surveys (Figure 5-6). Although the t test between reference and most disturbed lakes was
similar in both years, the positive relationship between disturbance and in lake level drawdown
was much less evident in the drier year (2007) than in 2012. In 2012 fewer than 5% of reference
lakes showed any drawdown at all, whereas 75 to 95 % of reference lakes showed drawdown in
2007 - with a lot of overlap in the inter-quartile ranges of reference and most disturbed sites.
RVegQ_OE, LitCvrQ^OE, and LitRipCvQ_OE in sub-sets and sub-regions of the U.S. universally
showed the same pattern of response as the nation, with the mean of reference sites
significantly greater than those for most-disturbed sites (Table 5-9). Discrimination was
generally greater for RVegQ_OE and LitRipCvQ_OE than for LitCvrQ_OE or the drawdown
indices. Discrimination of these 3 indices was somewhat greater for natural lakes than for
reservoirs, but good in both. RVegQ_OE was strongly and clearly associated with disturbance
(RT_NLA12) in all regions and years except for NPL, and SPL in the NLA 2007 survey year.
LitCvrQ_OE was strongly related to disturbance class in the CPL and NPL, moderately related to
disturbance in the NAP, TPL (2012), SPL, and XER; and associations were with disturbance were
weakest in the SAP, WMT, and TPL (2007). LitRipCvQ_OE was strongly and clearly associated
with disturbance (RT_NLA12) in all regions and both years.
Fergus et al. (2020) examined differences in lake hydrologic variables between the 2007 and
2012 surveys, providing insight on the sensitivity of lake levels and water balance parameters to
inter-annual climate conditions. Between-year variation in water-level decline was greater on
natural lakes than human-made lakes, suggesting that natural lakes are more responsive to
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changes in weather. They reported less vertical drawdown in natural lakes in 2012 (a cooler,
wetter weather year) compared to 2007, whereas large drawdown persisted on human-made
lakes, particularly in western regions. Dam and outlet structures can significantly alter lake and
stream hydrology and potentially mask effects from climate or weather. Fergus et al. (2020,
2021) suggested, based on the 2007- 2012 changes in evaporative concentration and water
levels and an index of the potential for anthropogenic hydro-alteration, that water levels in
natural lakes levels may be more responsive to temperature and precipitation in a given year,
whereas water levels in human-made lakes may be more strongly influenced by water
management and indirectly by weather conditions, particularly in western U.S. regions. Fergus
et al. (2021) also showed evidence that in the wetter eastern regions of the U.S., water
management infrastructure is used to stabilize lake and reservoir water levels, whereas water
management for irrigation, hydropower, and water supply in the drier regions leads to greater
level fluctuation and drawdown.
5.7 Discussion
The NLA and other lake survey and monitoring efforts increasingly rely upon biological
assemblage data to define lake condition. Information concerning the multiple dimensions of
physical and chemical habitat is necessary to interpret this biological information and
meaningfully assess ecological condition. The controlling influence of littoral structure and
complexity on lake biota has been long recognized, and recent research highlights the roles of
habitat structural components like littoral woody debris in providing refuges from predation
and affecting nutrient cycling and littoral production. NLA field crews characterized lake depth,
water surface characteristics, bank morphology and evidence of lake level fluctuations, littoral
and shoreline substrate, fish concealment features, aquatic macrophytes, riparian vegetation
cover and structure, and human land use activities. These littoral and riparian physical habitat
measurements and visual observations were made in a randomized array of 10 near-shore
littoral-riparian plots systematically spaced along the shoreline of each sample lake. Metrics
describing a rich variety of lake characteristics were calculated from this raw data, and many of
these were determined with moderate precision in the national dataset. For the NLA, we
summarize this information with four integrative measures of lake condition, and one measure
of lake drawdown and lake level fluctuation: RDis_IX, incorporating measures of the extent and
intensity of near-shore human land and water use activities; RVegQ, incorporating the structure
and cover in three layers of riparian vegetation, including inundated vegetation; LitCvrQ, a
combined biotic cover complexity measure including large woody snags, brush, overhanging
vegetation, aquatic macrophytes, boulders, and rock ledges; and LitRipCvrQ, which combines
RipVegQ and LitCvrQ. The measure of lake level drawdown incorporates both horizontal and
vertical fluctuation, comparing them to the regional mean values observed in least disturbed
lakes and reservoirs.
We modeled expected values of RVegQ, LitCvrQ, and LitRipCvrQ and their divergence from
reference conditions in least disturbed lakes using regression-based O/E models. The precision
of these O/E indices was moderate to high and showed good discrimination between least
disturbed and most disturbed lakes nationally, and within ecoregions. These results show that,
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compared with least disturbed reference lakes, those with moderate or high human
disturbances in the same region have reduced cover and extent of multi-layered riparian
vegetation or natural wetlands. In addition, those with moderate or high disturbance generally
also have reduced snag, brush and emergent aquatic macrophyte cover. These results
complement the results of the NLA 2012 public report and those of Kaufmann et al. 2014b,
2014c), confirming our general expectation that near-shore wetland and multi-layered riparian
vegetation and abundant, complex fish concealment features foster native fish,
macroinvertebrate, zooplankton, and avian assemblage integrity, whereas extensive and
intensive shoreline human activities that reduce natural riparian vegetation and reduce littoral
cover complexity are detrimental to these biotic assemblages.
We believe that the metrics and indices derived from the NLA physical habitat field approach
and the O/E indices expressing their divergence from least disturbed reference conditions
describe ecologically-relevant characteristics of lake habitat with sufficient precision to evaluate
near-shore lake habitat structure in national, state, and ecoregional assessments. Their
association with gradients of human disturbance demonstrates that they also describe lake
attributes that are vulnerable to anthropogenic degradation and potential for productive
restoration through lake and land management.
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68
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NLA 2022 Technical Support Document - August 2024
USEPA. 2017a. 2017 National Lakes Assessment: Field Operations Manual. EPA 841-B-16-001.
U.S. Environmental Protection Agency, Washington, DC.
USEPA. 2017b. National Lakes Assessment 2012: Techical Report. EPA 841-R-16-114. U.S.
Environmental Protection Agency, Washington, D.C.
West, E., and G. Ruark. 2004. Historical evidence of riparian forests in the Great Plains and how
that knowledge can aid with restoration and management. USDA Forest Service / UNL
Faculty Publications. Paper 7. Digital Commons at University of Nebraska - Lincoln.
http://digitalcommons.unl.edU/usdafsfacpub/7.
Whittier, T. R., D. B. Halliwell, and S. G. Paulsen. 1997. Cyprinid distributions in northeast U.S.A.
lakes: Evidence of regional-scale minnow biodiversity losses. Canadian Journal of Fisheries
and Aquatic Sciences 54:1593-1607.
Whittier, T. R., S. G. Paulsen, D. P. Larsen, S. A. Peterson, A. T. Herlihy, and P. R. Kaufmann.
2002. Indicators of ecological stress and their extent in the population of Northeastern
lakes: a regional-scale assessment. Bioscience 52(3):235-247.
Whittier, T. R., S. G. Paulsen, D. P. Larsen, S. A. Peterson, A. T. Herlihy, and P. R. Kaufmann.
2002b. Indicators of ecological stress and their extent in the population of northeastern
lakes: a regional-scale assessment. Bioscience 52:235-247.
Zohary, T., and I. Ostrovsky. 2011. Ecological impacts of excessive water level fluctuations in
stratified freshwater lakes. Inland Waters 1:47-59.
Zar, J.H. 1999. Biostatistical Analysis, 4th ed. Prentice-Hall, Inc. New Jersey, USA.
69
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NLA 2022 Technical Support Document-August 2024
Table 5-1. NLA reference sites from combined 2007 & 2012 surveys.
Selected using consistent criteria (Alan Herlihy's P,T_NLA12_2015, choosing 2012 visit for sites sampled in both
years). Bold font indicates grouping of reference sites used for modeling expected values for RVegQ, LitCvrQ, and
LitRipCvrQ.
EC09
ECOdS
Total
2007
2012
MAP
APPAL
67
23
44
SAP
APPAL
31
14
17
APPAL
(98)
(37)
(61)
CPL CPL 28 5 23
UMW UMW 49 18 31
TPL
CENPL
23
7
16
NPL
CENPL
11
3
8
SPL.
CENPL
35
21
14
CENPL
(69)
(31)
(38)
WMT
WEST
74
29
45
XER
WEST
20
4
16
WEST
(94)
(33)
(61)
Totals for lower 48 states
338
124
214
Table 5-2. Assignment of riparian vegetation cover complexity, littoral cover complexity, and littoral-riparian
habitat complexity index variants by aggregated ecoregion.
Aggregated
Omernik
Ecoregion
Riparian Vegetation
Cover Complexity index
(RVegQ)
Littoral Cover
Complexity Index
( LitCvrQ)
Littoral-Riparian
Habitat Complexity
Index
(LitRipCvrQ)
CPL
SAP
NAP, UMW
TPL, NPL, SPL
WMT, XER
RVegQ_2
RVegQ_2
RVegQ_2
RVegQ_7
RVegQ_8
LitCvrQ_b
LitCvrQ_c
LitCvrQ_d
LitCvrQ_d
LitCvrQ_d
LitRipCvrQ_2b
LitRipCvrQ_2c
LitRipCvrQ_2d
LitRipCvrQ_7d
LitRipCvrQ_8d
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NLA 2022 Technical Support Document-August 2024
Table 5-3. Summary of regression models used in estimating lake-specific expected values of Lake Physical Habitat
variables RVegQx, LitCvrQx and LitRipCvrQx under least disturbed conditions.
See Appendix A for model details.
REGION v = RVeaQ
v = LitCvrQ
v = LitRipCvrtl
NAP Ly* =f(Lat, Lon, LkOrig, RDisIX,) Ly =f(L_LkArea, RDisIX)
(R2=23% RMSE=0.162L*
(R2= 12%, RMSE=0.281L)
Ly =f(Lat, Lon, LkOrig, RDisIX)
(R2=24% RMSE=0.168L)
SAP Ly =f(Lonj
(R2=16% RMSE=0.119L)
Ly =f(ElevXLon, RDisIX)
(R2=19% RMSE=0.267L)
Ly =f(Lon, ElevXLon, Eievj
(R2=31% RMSE= 0.148L)
CPL y = f(ElevXLat, RDisIX)
(R2=39%, RMSE=0 .0896)
y =f(L_Elev, RDisIX)
(R2=25%, RMSE= 0.174)
y =f( L_Elev, RDisIX)
(R2=44%, RMSE=0.093)
UMW Ly = (mean LRVegQ)
(R2=0%, RMSE=0.153L)
Ly = (mean LitCvrQ)
(R2=0%, RMSE=0.199L)
Ly = (mean LitRipCvrQ)
(R2=0%, RMSE=0 .115L)
CENPL Ly =f(hiiAg)
(R2=15%, RMSE=0.318L)
Ly =f(LkOrig, hiiAg)
(R2=9% RMSE=0.276L)
Ly =f(hiiAg)
(R2=15%, RMSE=0.233L)
WMT Ly =f(Lat, Elev, L_LkArea, LkOrigin) Ly =f(Lat, Elev, L_LkArea, LkOrigin) Ly =f(Lat, Elev, L_LkArea, LkOrigin)
(R2=28% RMSE=0.167L) (R2=16% RMSE=0.244L) (R2=29%, RMSE=0.145L)
XER Ly =f(Lat, Elev)
(R2=24%, RMSE=0.284L)
Ly =f(Lat, Elev)
(R2=16%, RMSE=0.290L)
Ly =f( Lat, Elev)
(R2=21%, RMSE=0.265L)
*Ly refers to Logio-transformed lake habitat metric values.
**L refers to RMSE's that are in Logio units (e.gv 0.162L)
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NLA 2022 Technical Support Document - August 2024
Table 5-4. Null Model Geometric Means (gMean), geometric Standard Deviations (gSD), 5th percentiles, and 25th
percentiles of habitat index values in least disturbed reference lakes in the aggregated ecoregions of the NLA.
The gMeans and gSDs are antilogs of mean and SD of logio-transformed index values (LogMean and LogSD). Bold,
italicized text identifies minimum LogSD and gSD values, i.e., the most precise models for each index. Bold,
underlined text marks the least precise models. gSDs calculated from log-transformed variables are expressions of
the proportional variance of these distributions, so are directly comparable among regions with different gMeans.
A range of+lLogSD is equivalent to multiplying and dividing the gMean by the gSD. For example, the gMean +1
gSD for the riparian vegetation cover complexity index in least disturbed NAP lakes translates to a range of RVegQ
from 0.182 to 0.338: the geometric mean habitat index value of 0.2482 multiplied and divided by 1.363. The 5th
and 25th percentiles were estimated, respectively, as the mean of log-transformed index values minus 1.65 and
0.67 times the SD of log-transformed habitat index values (see Table 5-2 for the variant of each index used). All
percentiles are expressed in the units of the habitat indices, i.e., as antilogs of log-transformed values. (Note that
the constant 0.01 is subtracted from all antilogs because it was added when O/E values were log-transformed).
Aggregated
Refo7i2
Refo7i2
Refo7i2
Refo7i2
Refo7i2
Refo7i2
eco region
Index
LogMean
LogSD
gMean
gSD
est 5th%
est 25th %
Riparian Veaetation Cover Complexity:
NAP null
RVegQ
-0.5881
0.1345
0.2482
1.363
0.1449
0.1998
SAP null
RVegQ
-0.6111
0.1277
0.2348
1.342
0.1407
0.1911
UMWnull
RVegQ
-0.6130
0.1533
0.2338
1.423
0.1262
0.1824
CPLnull
RVegQ
-0.6645
0.2810
0.2065
1.910
0.0644
0.1304
CENPLnull
RVegQ
-0.8346
0.3427
0.1364
2.201
0.0298
0.0760
TPLnull
RVegQ
-0.7295
0.3129
0.1764
2.055
0.0468
0.1050
NPLnull
RVegQ
-1.1352
0.2500
0.0632
1.778
0.0183
0.0398
SPLnull
RVegQ
-0.8093
0.3402
0.1451
2.189
0.0326
0.0817
WMTnull
RVegQ
-0.5900
0.1922
0.2470
1.557
0.1138
0.1811
XERnull
RVegQ
-0.8301
0.3070
0.1379
2.028
0.0360
0.0821
Littoral Cover Complexity:
NAPnull
LitCvrQ
-0.8174
0.2418
0.1423
1.745
0.0508
0.9049
SAPnull
LitCvrQ
-0.6469
0.2873
0.2155
1.938
0.0657
0.1347
UMWnull
LitCvrQ
-0.8756
0.1994
0.1232
1.583
0.0524
0.0879
CPLnull
LitCvrQ
-0.4883
0.2331
0.3049
1.710
0.1240
0.2167
CENPLnull
LitCvrQ
-1.0164
0.2880
0.0863
1.941
0.0222
0.0518
TPLnull
LitCvrQ
-0.9927
0.3190
0.0917
2.084
0.0203
0.0522
NPLnull
LitCvrQ
-0.9974
0.2116
0.0906
1.628
0.0350
0.0626
SPLnull
LitCvrQ
-1.0389
0.2929
0.0814
1.963
0.0200
0.0482
WMTnull
LitCvrQ
-1.0162
0.2578
0.0863
1.811
0.0262
0.0547
XERnull
LitCvrQ
-1.1457
0.2990
0.0615
1.991
0.0130
0.0351
Littoral-Riparian Habitat Complexity:
NAPnull
LitRipCvrQ
-0.6740
0.1404
0.2018
1.382
0.1143
0.1606
SAP null
LitRipCvrQ
-0.6069
0.1690
0.2372
1.476
0.1201
0.1805
UMWnull
LitRipCvrQ
-0.7083
0.1149
0.1857
1.303
0.1165
0.1541
CPLnull
LitRipCvrQ
-0.5391
0.1687
0.2796
1.475
0.1422
0.2128
CENPLnull
LitRipCvrQ
-0.8820
0.2508
0.1212
1.782
0.0406
0.0791
TPLnull
LitRipCvrQ
-0.8230
0.2813
0.1403
1.911
0.0416
0.0874
NPLnull
LitRipCvrQ
-1.0442
0.1887
0.0803
1.544
0.0341
0.0575
SPLnull
LitRipCvrQ
-0.8698
0.2305
0.1902
1.700
0.0462
0.0846
WMTnull
LitRipCvrQ
-0.7369
0.1677
0.1733
1.471
0.0869
0.1315
XERnull
LitRipCvrQ
-0.9455
0.2818
0.1034
1.913
0.0289
0.0634
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NLA 2022 Technical Support Document - August 2024
Table 5-5. 0/E Physical Habitat Model means (LogMean, gMean), standard deviations (LogSD, gSD), and percentiles
of the distribution of habitat index O/E values for least disturbed reference lakes in the aggregated ecoregions of
the NLA.
See Table 5-3 for the variant of each index used. The gMean and gSD are antilogs of mean and SD of logio-
transformed index values (LogMean and LogSD). Percentiles were estimated, respectively, as the log-transformed
index O/E value of 0.0 (see text) minus 1.65 and 0.67 times the SD of log-transformed habitat index values. Bold,
italicized text identifies minimum SD values, i.e., the most precise models for each index. Bold, underlined text
marks the least precise models. gSDs calculated from log-transformed variables are expressions of the proportional
variance of these distributions, so are directly comparable among regions with different geometric means. A range
of +1SD is calculated by multiplying and dividing the gMean by the gSD. For example, the LogMean + lLogSD for
the riparian vegetation cover complexity O/E index in least disturbed lakes of the NAP (0.04276 + 0.1255)
translates to a range of O/E values from 0.78 to 1.31: the geometric mean habitat index O/E value of 1.00 (antilog
of+0.04276 = 1.10 minus log-transform constant 0.10) multiplied and divided by 1.34, the antilog of 0.1255. All
percentiles expressed as antilogs of log-transformed values minus constant 0.10. We based physical habitat
condition criteria based on the distribution of O/E index values in least disturbed lakes within each region. The 5th
and 25th percentiles, respectively, were set as the upper bounds for poor and fair condition.
Aggregated
Ref 0/E
Ref 0/E
Ref O/E
Ref O/E
Ref O/E
Ref O/E
ecoregion
Index
LogMean
LogSD
gMean
gSD
5th %tile
25th %tile
NAP MLR Model
RVegCLOE
(-0.00811)
(0.1255)
(0.88)
(1.34)
NAPoEYint
// //
+0.04276
0.1255
1.00
1.34
0.5850
0.8092
SAP MLR Model
RVegCLOE
+0.04226
0.1105
1.00
1.29
0.6244
0.8295
UMWmlR Model
RVegCLOE
+0.0428
0.1442
1.00
1.39
0.5381
0.7835
CPL MLR Model
RVegQ^OE
(-0.0617)
(0.2113)
(0.87)
(1.63)
CPLoEYint
// //
-0.00067
0.2129
0.90
1.63
0.3449
0.6191
CENPL MLR Mode
RVegQ^OE
(-0.02799)
(0.3165)
(0.84)
(2.07)
| CENPLoEYint
// //
+0.04688
0.2928
1.01
1.96
0.2663
0.6091
WMTmlR Model
RVegQ^OE
+0.04290
0.1535
1.00
1.42
0.5162
0.7711
XERmlR Model
RVegQ^OE
+0.04199
0.2656
1.00
1.84
0.3016
0.6312
NAP MLR Model
LitCvrCLOE
(+0.04502)
(0.2330)
(1.01)
(1.71)
NAPoEYint
// //
+0.04665
0.2330
1.01
1.71
0.3594
0.6772
SAP MLR Model
LitCvrCLOE
(-0.05093)
(0.2500)
(0.79)
(1.78)
SAPoEYint
// //
+0.04287
0.2440
1.00
1.75
0.3368
0.6575
UMWmlR Model
LitCvrCLOE
+0.04422
0.1954
1.00
1.57
0.4245
0.7152
CPL MLR Model
LitCvrCLOE
(-0.03310)
(0.1909)
(0.83)
(1.55)
CPLoEYint
// //
-0.00743
0.1940
0.88
1.56
0.3704
0.6288
CENPL MLR Model
LitCvrCLOE
(+0.00495)
(0.2870)
(0.91)
(1.94)
CENPLoEYint
// //
+0.02752
0.2839
0.97
1.92
0.2624
0.5876
WMTmlR Model
LitCvrCLOE
+0.03770
0.2528
0.99
1.79
0.3174
0.6385
XERmlR Model
LitCvrCLOE
+0.03451
0.2983
0.98
1.99
0.2486
0.5834
NAP MLR Model
LitRipCvrQ^OE
(+0.00344)
(0.1321)
(0.91)
(1.36)
NAPoEYint
// //
+0.04230
0.1321
1.00
1.36
0.5672
0.7990
SAP MLR Model
LitRipCvrQ^OE
+0.04326
0.1329
1.00
1.36
0.5667
0.7999
UMWmlR Model
LitRipCvrQ^OE
+0.04199
0.1110
1.00
1.29
0.6252
0.8296
CPL MLR Model
LitRipCvrQ^OE
(-0.0248)
(0.1230)
(0.84)
(1.33)
CPLoEYint
// //
+0.01615
0.1234
0.94
1.33
0.5494
0.7580
CENPL MLR Model
LitRipCvrQ^OE
(-0.0121)
(0.2413)
(0.87)
(1.74)
| CENPLoEYint
// //
+0.04303
0.2246
1.00
1.68
0.3703
0.6808
WMTmlR Model
LitRipCvrQ^OE
+0.04200
0.1366
1.00
1.37
0.5556
0.7922
XERmlR Model
LitRipCvrQ^OE
+0.04012
0.2552
1.00
1.80
0.3159
0.6398
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NLA 2022 Technical Support Document-August 2024
Table 5-6. Empirical 75th and 95th percentiles of the distribution of vertical and horizontal drawdown.
As interpreted from indicators of lake levei fluctuation {bfxVertHeight and bfxHorizDist) at least disturbed
reference lakes sampled by NLA in 2007 and 2012. We used the 75th and 95th percentiles to define the boundaries
between small, medium and large magnitude of drawdown.
Number of Reference Lakes
(2007+2008)
Vertical Drawdown (m)
(bfxVertHeight)
Horizontal Drawdown (m)
(bfxHorizDist)
Ecogion
Lake
Origin
Total
Natural
Human-
made
median
75,h%
95th%
median
75th%
95th%
NAP
All
67
54
13
0.000
0.12
0.470
0.00
0.25
1.65
SAP
All
31
0
31
0.000
0.20
0.760
0.00
0.20
2.15
UMW
All
49
49
0
0.000
0.11
0.50
0.00
0.51
2.65
CPL
All
28
5
23
0.000
0.03
1.00
0.00
0.10
4.00
CENPL
Natural
29
29
0
0.000
0.06
0.28
0.00
0.10
2.85
u u
Human-
made
39/40
0
39/40
0.010
0.36
1.20
0.21
1.55
14.63
WEST
Natural
69
69
0
0.021
0.33
1.00
0.00
0.64
9.43
// II
Human-
made
25
0
25
0.232
1.05
2.00
0.27
4.39
11.37
74
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NLA 2022 Technical Support Document - August 2024
Table 5-7. Precision of the key NLA Physical Habitat indices used as the primary physical habitat condition
measures in the NLA.
NLA PHab Indices
Orep
RQobs
Orep/R Qobs
S/N
RDisJX
0.098
0.0-+0.950
0.103
9.1
L_ RVegQc
0.144
-2.0--0.266
0.083
6.6
L_RVegQc3OE
0.130
-1.0-+0.666
0.078
5.0
L_LitCvrQc
0.190
-2.0-+0.0266
0.094
3.4
L_LitCvrQc30E
0.188
-1.0-+0.759
0.107
2.2
L_LitRipCvrQc
0.134
-2.0--0.135
0.072
5.6
L_LitRipCvrQc30E
0.122
-1.0-+0.681
0.073
4.1
L_ VertDD
0.193 (0.266)
-1.0-+1.654
0.073 (0.100)
5.9 (2.7)
L_HorizDD
0.148 (0.283)
0.0-+2.873
0.052 (0.099)
11.0(3.8)
Precision is expressed as: 1) the pooled standard deviation of repeat visits (orep), 2) precision
relative to potential or observed range (orep/Rgpot and orep/Rgpot), and 3) the signal: noise
ratio, where signal is among-lakes variance and noise is within-lake variance during the same
year and season (S/N = o2lake/o2rep). Analysis was based on NLA field measurements on a
summer probability sample of 1203 lakes in the 48 conterminous U.S. states, with repeat
sampling on a random subset of 88 of those lakes during the summer of 2012. Six of the sample
lakes showed very large changes in water level, which affected the littoral and riparian indicator
values. We excluded these 6 lakes in this analysis, except for values within perentheses.
RDisJX is the Near-shore human disturbance index, RVegQc is the Riparian vegetation cover &
structure index, Log(RVegQc30E) is the log-transformed O/E index for Riparian vegetation
cover & structure, LitCvrQc is the Littoral cover complexity index, Log(LitCvrQc30E is the log-
transformed O/E index for Littoral cover complexity, LitRipCvrQc is the Littoral-riparian habitat
complexity index, Log(LitRipCvrQc30E) is the log-transformed O/E index for Littoral-riparian
habitat complexity, L_VertDD = LoglO(Vertical drawdown +0.1m), and L_HorizDD =
LoglO(Horizontal drawdown + lm).
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NLA 2022 Technical Support Document - August 2024
Table 5-8. Association of NLA-2012 Physical Habitat Indices with high and low anthropogenic disturbance stress
classes (RT_NLA12 = R and T), defined as least disturbed and most disturbed within NLA regions.
The t-values test the null hypothesis that the mean value of the habitat index in Reference sites minus the mean in
most disturbed sites was zero in the NLA 2012 survey. Positive tRTvalues indicate that habitat index values are
greater in least disturbed sites; negative values indicate higher index values in disturbed sites. See Figure 5-6 for
box and whisker plots by NLA regions, presented separately for the NLA 2012 and 2007 surveys.
* Note that RDis_IX was one of the screening variables used to define least disturbed reference
sites (RT_NLA12=R) and most disturbed sites (RT_NLA12=T), and was a very influential. The
drawdown variables bfxVertHeight and bfxHorizDist were also used in the screening process,
but had only a minor influence on the definition of sites.
NLA Physical Habitat Indices
tRT PRT>I tRT I
RDis_IX- Near-shore human disturbance index
L_RVegQc - Riparian vegetation cover & structure index
L_RVegQc30E - O/E index for Riparian vegetation cover & structure
L_LitCvrQc - Littoral cover complexity index
L_LitCvrQc30E- O/E index for Littoral cover complexity
L_LitRipCvrQc-Littoral-riparian habitat complexity index
L_LitRipCvrQc30E - O/E index for Littoral-riparian habitat complexity
L_VertDD - Logio(Vertical drawdown +0.1m)
L_HorizDD- Logio(Horizontal drawdown +1.0m)
-25* <0.0001*
13 <0.0001
14 <0.0001
8.3 <0.0001
9.3 <0.0001
13 <0.0001
14 <0.0001
-4.3* <0.0001*
-4.7* <0.0001*
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NLA 2022 Technical Support Document - August 2024
Table 5-9. Association of NLA 2007 and 2012 Physical Habitat Indices with high and low anthropogenic disturbance
stress classes (RT_NLA12 = L and M), defined as least disturbed and most disturbed within NLA regions.
The t-values test the null hypothesis that the mean value of the habitat index in Reference sites minus the mean in
most disturbed sites was zero in the Domain specified in column 1. Positive t-values indicate that habitat index
values are greater in least disturbed sites; negative values indicate higher index values in disturbed sites. See
Figure 5-6 for box and whisker plots by NLA regions, presented separately for the NLA 2012 and 2007 surveys.
DOMAIN
L_RVegOE
L_LitCvrOE
L_LitRipCvrOE
LJHorizDD
National
07&12
ig****
12****
ig****
_7 7****
National 07&12
Natural
Human-
14****
g g****
14****
-3.5***
made
12****
g g****
12****
-6 o****
National 2007
12****
-j 2****
12****
_g 2****
2012
14****
g 2****
1^****
_4 7****
APPAL 2007
g 4****
3.0***
4 4****
+1.9
2012
g 4****
^ 1****
4 1****
-3 2***
NAP 2007
4 Q***
2.4**
4 1***
+1.1
2012
2 g***
2 g***
4 2****
-2.4*
SAP 2007
4 g****
1.1
2.9**
-0.2
2012
g 2****
1.4
3.3**
-2.4*
CENPL 2007
4 4****
2.5**
^ Q****
_4 Q****
2012
g 2****
^ 5****
g 4****
-0.6
TPL 2007
4 Q***
0.3
2.9**
-1.2
2012
3.6***
3.3**
2 7***
0.6
NPL 2007
1.3
4.6***
4 g***
i****
2012
2.4*
2.4*
2.2*
+1.6*
SPL 2007
1.4
2.1*
2 2**
-1.2
2012
g Q****
4 4****
gi****
-2.2*
CPL 2007
4 5***
1.4
4 g****
-1.3
2012
3.6***
4 2****
^ 4****
-0.5
UMW 2007
g 5****
g 2****
-j 2****
+4 4****
2012
g i****
3.3***
g 5****
-0.5
WEST 2007
g y****
2 ^***
-j y****
_g i****
2012
g 2****
2 2***
7 2****
2****
WMT 2007
g 2****
1.6*
^ 4****
7****
2012
g y****
2.3*
g Q****
g****
XER 2007
g 2****
3.5***
^ g****
-4 g****
2012
4 5****
2.0*
3.6**
-1.4
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NLA 2022 Technical Support Document - August 2024
Near-Shore Station NLA-2007:
Near-Shore Station NLA-2012-present:
Observation station
Figure 5.1. Field sampling design with 10 near-shore stations at which data were collected to characterize near shore
lake riparian and littoral physical habitat in the 2007 and 2012 National Lakes Assessment (NLA) surveys. The 10
stations were systematically spaced around the shore of the lake from random starting point. Insert shows riparian
plot, shoreline band, littoral plot, and (for NLA 2012 and later) drawdown zone plot located at each station.
Littoral-Riparian Plot
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I
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1 WMT 2-UMW
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i r
8-CPL 9-NPL
Figure 5.2. Near-shore anthropogenic disturbance (RDisJX) in NLA0712 regions, ordered by their median
Reference site RDis.
Upper plot: Least disturbed reference sites. Lower plot: all sites. Unweighted sample statistics are shown; box
midline and lower and upper ends show median and 25th and 75th percentile values, respectively; whiskers show
maximum and minimum observations within 1.5 times the interquartile range above/below box ends; circles
show outliers.
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Figure 5.3, Near-shore anthropogenic disturbance in NLA0712 least disturbed reference sites (median RDisJX),
ordered by aggregated region according to the same median level of near-shore disturbance.
The NLA EC09 regions NPL, SPL, and TPL are combined into the Central Plains (CENPL) region.
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NLA 2022 Technical Support Document - August 2024
Log(RVegQ):
Log(LitCvrQ):
RVegLSDrNul
RVegLSDrOEaj
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| O LtRpCvLSDrNul • LtRpCvLSDrOEaj |
Figure 5.4. LogSD's for Null-Model and regression-based O/E model for Near-shore RVegQ, LitCvrQ, and
LitRipCvrQ in the set of least disturbed lakes and reservoirs (
Table 5-1) sampled in the combined NLA 2007 and 2012 surveys.
X-axis shows the 7 modeling regions ordered by increasing median RDisJX in the reference sites. The
NLA EC09 regions NPL, SPL, and TPL are combed into the Central Plains (CENPL) region. Low variance
among reference sites denotes greater precision in estimating expected reference condition. The
smaller variance in regression-based O/E models (black dots) illustrate their greater precision compared
with null models (ooen circles) for a given indicator and region.
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Figure 5.5. Contrasts in key NLA physical habitat index values among least disturbed reference (L),
intermediate (I), and most disturbed (M) lakes in the contiguous 48 states of the U.S. based on combined NI_A
2007 and 2012 data. Unweighted sample statistics are shown; box midline and lower and upper ends show
median and 25th and 75th percentile values, respectively; whiskers show maximum and minimum
observations within 1.5 times the interquartile range above/below box ends; circles show outliers. See Table
5-9 for t and p values for the differences between means for least disturbed reference (L) and most disturbed
(M) sites.
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Figure 5.6, Contrasts in key NLA physical habitat index values among least disturbed reference (L), intermediate (I),
and most disturbed (M) lakes in the contiguous 48 states of the U.S. shown separately for the NLA 2007 and 2012
surveys.
Unweighted sample statistics are shown; box midline and lower and upper ends show median and 25th and 75th
percentile values, respectively; whiskers show maximum and minimum observations within 1.5 times the
interquartile range above/below box ends; circles show outliers. See Table 5-9 for t and p values for the differences
between means for reference (L) and most disturbed (M) sites.
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Chapter 6: Water Chemistry
6.1 Background information
The NLA public report presents the percentage of lakes in different condition class categories
for water quality stressor data collected at the deepest part of each study lake. Field sampling
included a depth profile and a 0-2 m depth integrated water sample. Variables analyzed and
presented in the NLA 2022 report include: total nitrogen (TN), total phosphorus (TP),
chlorophyll a (CHLA), acidity, dissolved oxygen, and atrazine. Turbidity data were also reviewed
but are not presented in the public report. Acidity, dissolved oxygen, trophic state class, and
atrazine benchmarks were based on established criteria and applied consistently across the
nation. Good, fair and poor condition classes were established for TP, TN, and CHLA using the
percentile of reference sites approach used in prior NLAs (Herlihy and Sifneos, 2013). Separate
benchmarks were established for each of the nine ecoregions. The benchmarks used in the
2022 analyses are consistent with those developed for the 2017 survey. In NLA 2017, the
benchmark values were revised; therefore, direct comparisons should not be made between
2007 and 2012 condition class results and those reported in 2017 and 2022. Human health
water quality indicators (i.e., cyanotoxins and enterococci) are discussed in Chapter 8.
6.2 Chemical condition benchmarks
6.2.1 Acidity
For setting acidity classes, concentrations of acid neutralizing capacity (ANC) and dissolved
organic carbon (DOC) were analyzed following the scheme developed by Herlihy et al. (1991).
Sites with acid neutralizing capacity (ANC) > 50 ueq/L were considered to be non-acidic and
least disturbed (good condition class) for acidification. Sites with ANC < 50 |-ieq/L and DOC
values > 6 mg/L were classified as naturally acidic due to organic acids (also good condition
class). Sites with ANC < 0 |-ieq/L and DOC values < 6 mg/L were classified as acidic due to either
acidic deposition or acid mine drainage and considered most disturbed or poor condition class.
Sites with ANC between 0 and 50 |-ieq/L and DOC < 6 mg/L were considered acid-influenced but
not currently acidic. These low ANC sites typically become acidic during high flow events
(episodic acidity) and were considered moderately disturbed (fair condition class).
6.2.2 Dissolved Oxygen
Depth profiles of dissolved oxygen were collected at the deepest location of the lake. Surface
water dissolved oxygen was calculated by removing all duplicate depth observations and taking
the mean of all dissolved oxygen values between 0 and 2 meters depth, inclusive. If the lake
was shallower than 2 m depth, the entire depth profile was used. Mean surface water dissolved
oxygen was classified into three classes, good (>5 mg/L), fair (3-5 mg/L), and poor (<3 mg/L).
Dissolved oxygen benchmarks of 5 mg/L and 3 mg/L represent US EPA's dissolved oxygen water
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NLA 2022 Technical Support Document - August 2024
qualtiy criteria recommendations for a warmwater daily minimum for early life stages and
other life stages, respectively (USEPA 1986).
6.2.3 Trophic State
Lakes have long been classified according to their trophic state. By the dictionary, "trophic" is
defined as of or relating to nutrition. A eutrophic lake has high nutrients and high algal and/or
macrophyte plant growth. An oligotrophic lake has low nutrient concentrations and low plant
growth. Mesotrophic lakes fall somewhere in between eutrophic and oligotrophic lakes and
hypereutrophic lakes have very high nutrients and plant growth. Lake trophic state is typically
determined by a wide variety of natural factors that control nutrient supply, climate, and basin
morphometry. Trophic state can be defined based on a number of different nutrient or plant
biomass variables. For NLA, trophic state was defined using concentrations of CHLA (Table 6-1)
The same trophic state classification was used for all ecoregions.
Table 6-1. Trophic State Classification used in NLA
Analyte
Oligotrophic
Mesotrophic
Eutrophic
Hypereutrophic
Chlorophyll a (|ig/L)
<2
>2 and <7
>7 and <30
>30
6.2.4 Total nitrogen, total phosphorus, chlorophyll a, and turbidity
TN, TP, CHLA, and turbidity were classified into good, fair and poor condition classes based on
percentiles of the nutrient reference site distribution (Herlihy and Sifneos, 2008, 2013).
Because nutrients (TN, TP) were used to select biological reference sites, the biological
reference sites could not be used as is for nutrient reference lakes due to circularity. The same
nutrient benchmarks used in NLA 2017 were used in NLA 2022. In 2017, to develop nutrient
reference sites, we compiled all sampled sites in NLA 2007, 2012, and 2017 as was done for the
biological reference condition process (see Chapter 3:). All sites were then passed through the
NLA biological reference screening process for their ecoregion as described in section 3.4 with
one exception. To avoid complete circularity, TP and TN thresholds were removed as screening
variables in the screening process.
After this initial screening, there remained a fairly strong disturbance signal in the reference
sites as evidenced by looking at relationships with GIS landscape stressor variables in particular,
% Agriculture watershed and % Developed watershed. In order to remove this disturbance
signal, an additional GIS stressor screen was added to the process to remove from the nutrient
reference site pool those sites that failed the filtering for these two metrics. For watershed %
agriculture, ecoregional criteria were used: >10% for NAP, WMT, and XER lakes; >25% for NPL,
SAP, SPL, and UMW lakes; >40% for CPL lakes; and >50% for TPL lakes. For watershed %
developed, a >10% criterion was used for all ecoregions except the CPL where a >15% filtering
criterion was used.
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NLA 2022 Technical Support Document - August 2024
For calculating the nutrient condition class benchmarks used in the NLA 2017 and 2022 public
reports, we used these 2007-2017 all NLA nutrient reference sites sampled from 2007-2017
(Table 6-2). When a site was sampled multiple times, only the first visit to the most recent year
of sampling was used to calculate percentiles so reference sites were not double-counted.
Before calculating benchmarks, a 1.5*IQR outlier analysis was done on the reference site
concentrations to remove outliers. Separate benchmarks were calculated for each of the nine
NARS ecoregions (Fig. 3-1). In addition,, and just in the Southern Plains, separate benchmarks
were calculated for natural and manmade lakes due to large differences in least disturbed
nutrient concentrations separately. Thresholds were determined for TP, TN, CHLA, and
turbidity. The cutoff between good and fair condition class was set at the 75th percentile (Q3) of
reference lakes, and the cutoff between fair and poor condition class was set at the 95th
percentile (P95) of reference lakes (Table 6-3).
Table 6-2. Number of unique nutrient reference sites used to calculate nutrient benchmarks (2007-2017 data).
Ecoregion
Number of Nutrient Reference
Sites
CPL
33
NAP
88
NPL
16
SAP
41
SPL-manmade
24
SPL-natural
20
TPL
26
UMW
87
WMT
142
XER
32
Total
509
There was a very large difference in the absolute concentrations of TP and TN among
ecoregions in the nutrient reference sites (Figure 6-1 and Figure 6-2). Looking at the data, it is
also evident why the natural lakes in the SPL need their own benchmark versus human-made
SPL lakes. Table 6-3 reports the 75th and 95th percentile-based benchmarks used to define the
good, fair and poor condition classes for TP, TN, CHLA, and turbidity for each of the ecoregions
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1000
3-
U5
2
it£
.1
1000
100
10
CPL
NAP
NPL
SAP SPLrnan SPLnat TPL UMW WMT XER
NLA Ecoregion
Figure 6.2. Box and whisker plot of Total Nitrogen in GIS screened, outlier removed, 2007-2.017 nutrient reference
sites by ecoregion. Boxes are the interquartile range, whiskers are 5th/95th percentiles.
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Table 6-3. NLA 2017 good, fair, and poor benchmarks (75th/95th percentiles) for TP, TN, CHLA, and turbidity
condition classes.
TP (Hg/L)
TP (|ig/L)
TN (Hg/L)
TN (|ag/L)
75th
95th
75th
95th
Ecoregion
Good-fair
Fair-poor
Good-fair
Fair-poor
CPL
43.0
59.5
659
923
NAP
16.0
27.9
428
655
NPL
63.0
82.0
849
1,620
SAP
18.0
33.0
266
409
SPL-
30.0
43.0
650
830
manmade
SPL-natural
486
839
7,840
11,100
TPL
38.4
57.5
865
1,350
UMW
24.8
40.0
766
926
WMT
23.4
43.0
253
429
XER
44.0
84.8
605
954
CHLA (|ag/L)
CHLA (|ag/L)
Turbidity (NTU)
Turbidity (NTU)
75th
95th
75th
95th
Ecoregion
Good-fair
Fair-poor
Good-fair
Fair-poor
CPL
12.7
28.0
3.42
4.15
NAP
4.52
8.43
1.30
2.52
NPL
10.9
19.3
3.08
4.46
SAP
5.54
13.1
2.83
4.21
SPL-
8.97
12.6
3.32
4.67
manmade
SPL-natural
118
219
71.3
86.4
TPL
13.9
19.8
3.64
4.23
UMW
7.43
14.6
2.18
3.32
WMT
1.86
3.86
0.910
1.60
XER
5.92
9.00
2.97
4.84
6.2.5 Atrazine
Atrazine water chemistry analyses were added to the NLA in 2012. Samples for atrazine were
collected using a 0-2 m vertically integrated water column sampler at the open-water site.
Measured concentrations were compared to nationally consistent benchmarks to estimate
ecological risk. The NLA also reports on the percentage of lakes with detections and changes in
detection over time. Detection is defined as a value greater than the minimum detection limit
(MDL). When the MDL changed between surveys, the greatest MDL for all surveys was used to
determine detection.
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The NLA atrazine benchmark is the EPA's aquatic plant concentration equivalent level of
concern (CE-LOC) used in the EPA's atrazine ecological exposure monitoring program. This
benchmark ensures that atrazine levels will not cause significant changes in aquatic plant
community structure, function and productivity (US EPA Atrazine website). In NLA 2012, the
EPA used a proposed CE-LOC of 4 ppb for atrazine risk results. In NLA 2017, this value was
updated to the current CE-LOC of 3.4 ppb. To report on the percentage of lakes with atrazine
detections, a consistent detection value was selected. The MDL was equal to 0.046 ppb for
most samples in NLA 2012 and 0.03 ppb for most samples in NLA 2017 and 2022. Therefore,
detection results in the public report and data dashboard present the percentage of lakes with
measured values greater than or equal to 0.046 ppb for all surveys.
6.2.6 Within-year variability
To examine within-year variability of water chemistry data, analysts used the revisit sites from
the NLA 2007 and 2012 (2,482 sites with 192 sites with revisits) to calculate S:N estimates for
the water chemistry indicators presented in Table 6-2 Overall S:N and pooled standard
deviation (SD) for NLA 2007 and 2012 surface water chemistry within three concentration range
classes.Table 6-2. Metrics with high S:N are more likely to show consistent responses to human
caused disturbance, and S:N values < 1 indicate that sampling a site twice yields as much or
more metric variability as sampling two different sites (Stoddard et al., 2008).
Table 6-2 Overall S:N and pooled standard deviation (SD) for NLA 2007 and 2012 surface water chemistry within
three concentration range classes. N = 192
Parameter
S:N
Low
Medium
High
range
SD
Range
SD
range
SD
ANC
98.3
<500 ueq/L
28.9
500-2500
ueq/L
153
>2500 ueq/L
309
Chloride
78.7
0-250 ueq/L
9.32
250-1000
ueq/L
59.1
>1000 ueq/L
373
Chlorophyll-a
3.85
0-10 ug/L
2.47
10-50 ug/L
16.9
>50 ug/L
63.6
Color
8.2
0-10 PCU
4.32
10-50 PCU
5.9
>50 PCU
40.1
Conductivity
134
0-100 uS
6.13
100-500 uS
21.3
500-2000 uS
67
DOC
97.2
0-5 mg/L
0.388
5-10 mg/L
0.687
>10 mg/L
4.35
ph
5.44
0-6
0.111
6-8
0.28
>8
0.343
Sulfate
238
0-250 ueq/L
13.3
250-1000
ueq/L
50.2
>1000 ueq/L
364
Total Nitrogen
23.2
0-250 ug/L
42.5
250-1000 ug/L
160
>1000 ug/L
818
Total
Phosphorus
18.6
0-25 ug/L
5.24
25-100 ug/L
16.8
>100 ug/L
123
Turbidity
6.69
0-5 NTU
1.1
5-25 NTU
6.85
>25 NTU
33.9
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Within-year sampling variability for atrazine was assessed by comparing NLA, 2012, 2017 and
2022 visit 1 and 2 condition categories and is presented in Table 6-3Table 8-1. For atrazine
detection, results showed agreement in 238 (81%) of the 293 revisit sites. For atrazine risk
condition, 291 (99%) of the 293 risk categores were in agreement.
Table 6-3. Atrazine detection (a) and risk condition (b) contingency tables. N = 293
a)
Atrazine Detection
Visit 1
Detected
Not-detected
Not Assessed
Visit 2
Detected
64
30
Not detected
24
174
Not Assessed
1
b)
Atrazine Risk Condition
Visit 1
At or Below Benchmark
Above Benchmark
Not Assessed
At or Below Benchmark
289
2
Visit 2
Above Benchmark
1
Not Assessed
1
6.3 Literature cited
Carlson, R.E. 1977. A trophic state index for lakes. Limnology and Oceanography. 22:361-369.
Herlihy, A. T., P. R. Kaufmann, and M. E. Mitch. 1991. Chemical characteristics of streams in the
Eastern United States: II. Sources of acidity in acidic and low ANC streams. Water
Resources Research 27:629-642.
Herlihy, A. T., and J. C. Sifneos. 2008. Developing nutrient criteria and classification schemes for
wadeable streams in the conterminous USA. Journal of the North American
Benthological Society 27:932-948.
Herlihy, A. T., N. C. Kamman, J. C. Sifneos, D. Charles, M. D. Enache, and R. J. Stevenson. 2013.
Using multiple approaches to develop nutrient criteria for lakes in the conterminous
USA. Freshwater Science 32:367-384. doi: 10.1899/11-097.
US EPA. 1986. Quality Criteria for Water ("Gold Book"). EPA 440/5-86-001
US EPA. 2019. Recommended human health recreational ambient water quality criteria or
swimming advisories for microcystins and cylindrospermopsin. US EPA Office of Water.
EPA 822-R-19-001.
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Chapter 7: Zooplankton
7.1 Background information
Zooplankton assemblages have several attributes that make them potentially useful for
assessing the ecological condition of lakes (Stemberger and Lazorchak 1994, Jeppesen et al.
2011). Zooplankton are typically the dominant pelagic consumer in lakes (in terms of both
biomass and numbers (Larsen and Christie 1993). Taxa richness tends to be high in nearly all
lakes. Zooplankton species or guild structure can respond to abiotic stressors such as
eutrophication and acidification, and possibly climate change. Zooplankton occupy an
intermediate level in the overall food web of lakes, and thus can respond to stress responses
from within lower (e.g., phytoplankton) or higher trophic levels (e.g., fish). Zooplankton taxa
demonstrate a range of life history strategies and patterns (e.g., parthenogenesis, resting eggs)
that can be related to environmental stress, both natural and anthropogenic.
The use of zooplankton assemblages in the context of bioassessment appears to be limited,
with many studies focused mainly on taxa richness and taxonomic composition changes in
response to disturbance. Gannon and Stemberger (1978) discussed the potential of using
zooplankton communities to help determine trophic state in lakes, primarily through the use of
"indicator species" that were associated with either oligotrophic or eutrophic conditions.
Sprules and Holtby (1979) and Sprules (1980) examined the utility of using metrics related to
body size and feeding ecology of zooplankton to evaluate lake condition. Duggan et al. (2001,
2002) investigated the potential for developing bioindicators of trophic state using rotifer
assemblages. Dodson et al. (2005) concluded that zooplankton assemblages are indirectly
associated with land use through effects on riparian vegetation and lake characteristics such as
typology and water chemistry. Dodson et al. (2009) examined changes in zooplankton
community structure within a set of lakes in northern Wisconsin in relation to a variety of
within-lake and watershed level characteristics (including human disturbance in the riparian
zone). Stemberger and Lazorchak (1994) calculated 14 metrics based on taxonomy, body size,
life history stage, and trophic guild in 19 lakes in the Northeastern USA representing a gradient
of human disturbance, lake type, and land use. Stemberger and Miller (1998) discussed
expected changes in zooplankton assemblage trophic structure and species composition in
response to changes in the N:P ratio that might result from increased anthropogenic
disturbance.
More recently, there have been attempts to develop indices of biotic condition in lakes using
plankton assemblages, following two approaches. The multimetric approach pioneered by Karr
(e.g., Karr 1981, Karr 1991) has been implemented successfully for other assemblages (e.g., fish,
benthic invertebrates) in streams. Kane et al. (2009) combined zooplankton and phytoplankton
metrics from Lake Erie into a single multimetric index (MMI), the Planktonic Index of Biotic
Integrity, to reflect the response of the plankton to eutrophication. The second approach
(predictive model approach) compares the observed taxa collected at each site to the list of
taxa expected at that site under least disturbed conditions by means of an Observed/Expected
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index (0/E, e.g., Wright 1995, Hawkins et al. 2000, Hawkins 2006, Hawkins et al. 2010). The
predictive modelling approach has been used successfully for other assemblages, principally
benthic invertebrates, but also fish, in streams. The 2007 National Lake Assessment (NLA 2007)
used an O/E model that combined zooplankton and phytoplankton assemblages to assess
ecological condition of lakes in the conterminous US (Yuan et al. 2008, USEPA 2009). Table 7-1
summarizes current knowledge regarding the hypothesized responses of zooplankton
assemblages to different types of disturbance.
For NLA 2012, we decided to develop a MM I for pelagic zooplankton assemblages to assess
biological condition in lakes. We followed the approach described by Stoddard et al. (2008) to
screen candidate metrics for possible inclusion in an MMI. We then computed a large number
of MMIs based on all possible combinations of the metrics that passed the screening process,
following Van Sickle (2010), and selected the MMI that showed the best combination of
responsiveness to disturbance, repeatability, and low redundancy among component metrics.
For NLA 2022, we used the same MMIs to assess lake condition. This chapter provides
corrections and clarifications to the 2017 technical report that we identified for the NLA 2022
analyses.
7.2 Methods
7.2.1 Field methods
Sample collection procedures for zooplankton are described in the NLA 2022 FOM (USEPA
2022a). Field crews collected two samples at the index site (deepest area of a lake or the
midpoint of a reservoir) of each lake. The crew collected a "Coarse" sample (ZOCN) using a 1-m
long, 30-cm diameter plankton net with a reducing collar (20-cm diameter) having a mesh size
of 150 |am. The crew collected a "Fine" sample (ZOFN) using a 1-m long net with a reducing
collar (20-cm diameter) having a mesh size of 50 |am. The total tow length for each net was 5 m,
with the number of tows being dependent on the site depth. At lakes deeper than 6 m, a single
5 m vertical tow was done. At lakes between 4 and 6 m deep, two 2.5-m vertical tows were
done. At lakes between 2 to 3 m deep, five 1-m vertical tows were done. At lakes less than 2 m
deep, ten 0.5 m vertical tows were collected. Results from pilot studies suggested that a total
tow length of 5 m would provide sufficient numbers of taxa and organisms to develop the MMI
from nearly all lakes.
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Table 7-1. Hypothesized responses of zooplankton assemblages to disturbance
Hypothesized
Type of disturbance
Assemblage component or metric
response
References
Catchment development
Biomass of small cladocerans
Increase
Gelinas and Pinel-Alloul
(2008), Beaver et al. (2014)
Catchment development
Abundance of small daphnids and
cladocerans
Increase
Gelinas and Pinel-Alloul
(2008), Dodson et al. (2009),
Van Egeren et al. (2011),
Beaver et al. (2014)
Nutrients; Agricultural
Species richness
Decrease
Gannon and Stemberger
land use; riparian buffer
(1978), Dodson etal. (2005)
presence
Nutrients, land use
Large-sized species richness (e.g.,
Daphnia spp., calanoid copepods)
Decrease
Stemberger and Lazorchak
(1994)
Nutrients, land use
Small-sized species richness (e.g.,
Ceriodaphnia, rotifers)
Increase
Stemberger and Lazorchak
(1994)
Nutrients
Proportion of calanoid copepod
taxa
Decrease
Jeppesen et al. (2000), Du et
al. (2015)
Nutrients
Proportion of cyclopoid copepod
taxa
Increase
Jeppesen et al. (2000), Du et
al. (2015)
Nutrients
Ratio of calanoid copepods to
(cyclopoid copepods + cladocerans)
Decrease
Gannon and Stemberger
(1978), Kane et al. (2009)
Nutrients
Mean size
Decrease
Gannon and Stemberger
(1978)
Nutrients
Total biomass
Increase
Gannon and Stemberger
(1978)
Nutrients
Proportion of cladoceran biomass
Decrease
Jeppesen et al. (2000), Du et
al. (2015)
Nutrients
Relative abundance of calanoid
copepods
Decrease
Brooks (1969), Gannon and
Stemberger (1978)
Nutrients
Relative abundance of cyclopoid
copepods and small-bodied
cladocerans
Increase
Brooks (1969), Attayde and
Bozelli (1998)
Nutrients
Omnivorous taxa richness,
abundance, or biomass
Increase
Stemberger and Lazorchak
(1994), Stemberger et al.
(2001)
Nutrients (total P)
Biomass of rotifers and cyclopoid
copepods
Increase
Du et al. (2015)
Nutrients (total P)
Biomass of cladocerans and
cyclopoid copepods
Decrease
Du et al. (2015)
Nutrients, chlorophyll a,
Rotifer assemblage composition
Change
Duggan et al. (2001), (2002)
Secchi transparency,
temperature, dissolved
oxygen
Decrease in acid
Abundance of large-bodied
Decrease
Tessier and Horwitz (1990)
neutralization
zooplankton
capacity/calcium
concentrations
Invasive species
Native species richness, abundance,
or biomass
Decrease
Kane et al. (2009)
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7.2.1 Laboratory methods
Laboratory methods for zooplankton samples are described in the NLA 2022 laboratory
operations manual (USEPA 2022b). For both the ZOCN and ZOFN samples, the objective was to
subsample a sufficient volume to enumerate and identify at least 400 individuals. In the ZOCN
samples, only cladocerans and copepods (including copepedids) were enumerated. In the ZOFN
samples, only "small" taxa were enumerated (cladocerans < 0.2 mm long, copepods < 0.6 mm
long, rotifers, and nauplii). Veligers were not enumerated in the ZOFN sample. Individuals were
identified to species where possible. A "Large/Rare" search of the entire subsample was done
to identify larger taxa (e.g., Chaoborus, Leptodora, Mysidae, Ostracoda, and Hydracarina). In
2012, only the presence of these taxa in the subsample was noted (i.e., they were not
enumerated). In 2022, the laboratory recorded the number of organisms encountered in the
Large/Rare search.
Besides the number of individuals enumerated in the subsample (abundance), we estimated
the volume of water sampled by the tow using the tow length and the radius of the net mouth
for the sample. We used this tow volume to estimate density (no. individuals/L) of each taxon:
Density = ¦
Sample Vol. (mL)
- x Abundance
Vol. Counted (mL)
Tow Vol. (L)
The biomass (|ag dry mass/L) of each taxon in a sample was estimated by measuring the length
of 20 individuals (if possible). Length was converted to a biomass factor (|ag dry
mass/individual) based on published length-weight relationships (Dumont et al. 1975, McCauley
1984, Lawrence et al. 1987). Biomass was then calculated as:
/indiv\ r \ig \
Biomass = Density x Biomass Factor —-—
V L J \indiv'
In 2012, one laboratory did not estimate biomass for their samples. For these samples, we
estimated biomass as the mean biomass of a taxon from samples collected from surrounding
states or used a national mean (all samples collected that included the taxon) if the regional
sample size was too small. In 2022, one laboratory processed all zooplankton samples and
provided quantitative biomass data. This laboratory was different than the laboratory that
processed the NLA 2012 and 2017 samples. Biomass was estimated at each site for taxa that
had existing length-weight relationships, but estimates were not provided for all taxa. For sites
where a taxon with a missing biomass factor comprised less than 5% of the total number of
individuals counted, it was left as missing. For the remaining cases, we used available biomass
factors for that taxon from other sites (including NLA 2022, NLA 2017, and NLA 2012) from the
state and surrounding states to calculate a mean value to use.
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NOTE: In 2021, we discovered an error in the calculation of the tow volumes for the coarse-net
samples, which affected the values for density, biomass, any metrics based on either of these,
and any MMIs that included metrics based on density or biomass for both 2012 and 2017. The
2012 and 2017 data were corrected and republished to the NARS website.
7.3 Data preparation
7.3.1 Data quality assurance
We reviewed field data to correct recording errors and, when possible, to fill in missing values,
especially for critical variables like tow length. We reviewed the raw count files from each
laboratory to correct spelling errors in taxon names, and to make the taxonomy consistent
across laboratories (using the national lab taxonomy as the standard for all labs). We used
range checks on count, density, and biomass estimates to identify outliers, and corrected them
if they were due to recording errors. The number of errors discovered in the NLA 2022 field and
laboratory data was very low.
7.3.2 Master taxa list
We developed a master taxa list that included all taxa identified in the ZOFN and ZOCN samples.
The master taxa list included taxonomic information (e.g., phylum, class, order, suborder,
family, subfamily, genus, species, and subspecies). Autecological information for each taxon
included feeding guild (Predator, Omnivore, or Herbivore), Cladocera size class (LARGE vs.
SMALL), based primarily on data from Stemberger and Lazorchak (1994) and the Northeastern
Lakes Survey (Whittier et al. 2002), and a size class variable (NET_SZECLS_NEW) based on
whether a taxon was collected in the ZOCN samples vs. only in the ZOFN samples.
The laboratory identified 535 unique taxa in the NLA 2012 ZOCN and ZOFN samples
(variable=TAXANAME). We combined some of these unique taxa using a different variable
(TARGET_TAXON), which resulted in 481 unique taxon names as used in metric calculations.
We also had some information regarding non-native zooplankton taxa based on the USGS
Nonindigenous Aquatic Species (NAS) database (Fuller and Neilson 2015). Bosmina coregoni (or
Eubosmina coregoni), Cercopagis pengoi, Daphnia lumholtzi, Sinocalanus doerri,
Pseudodiaptomus forbesi, and Arctodiaptomus dorsalis were considered to be introduced to
North America. Eutymora affinis was considered to be introduced to inland lakes of the US.
Skistodiapomus pallidus was considered to be introduced to lakes in states outside of the
Mississippi-Missouri-Ohio River basins. EPA also reviewed the laboratory results for non-target
taxa. Non-target taxa are excluded from enumeration and are listed in Appendix D.
For NLA 2022, we updated the master taxa list from NLA 2017 to add new taxa and associated
autecological information that were identified in the coarse and fine net samples collected in
2022. The NLA 2022 taxa list for zooplankton contains 634 unique names (excluding the taxa
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listed in Appendix D) for the variable TARGET_TAXON, which are used for metric calculations.
This is an increase of 54 taxa from those included in the taxa list for NLA 2017.
7.3.3 Aggregations and rarefaction of count data
We aggregated some values of TARGET_TAXON within a given ZOCN or ZOCN sample. We
combined copepodites and nauplii with adults of the same taxon if both were present in a
sample. If a species and a lower level taxon (i.e., subspecies, variety, or form) were both
present in a single sample, we aggregated the count data to the species level.
After aggregating at the sample level, we combined the results for each ZOCN and ZOFN sample
to create a separate site-level count file. We assumed that individuals collected in the ZOCN
samples that were also present in the ZOFN sample represented smaller individuals that passed
through the coarse-mesh net, and so we added the counts from the two samples together.
Because not all zooplankton individuals in a sample can be confidently identified to species,
there is a risk of overestimating taxa richness. For each sample, we reviewed the list of taxa to
determine whether they were represented at more than one level of resolution. For example, if
a "Daphnia sp." was collected, and it was the only representative of the genus in the sample (or
at the site), we assigned it as distinct. If any other members of the genus were collected, then
we considered the unknown as not distinct. We used only the number of distinct taxa in the
sample to calculate any metrics based on species richness. We calculated distinct taxa for both
the sample-level aggregated count file and the site-level count file. Taxa that were identified
(but not enumerated) during the Large/Rare search were included in calculating richness
metrics.
Even with a fixed count subsampling approach, taxonomic richness and metrics can be
influenced by the number of individuals enumerated in a subsample (Stoddard et al. 2008). We
created an additional count file to use for metric calculation by subjecting the sample-level
aggregated count data to a rarefaction procedure to randomly select 300 individuals per
sample (for those samples that had > 300 individuals enumerated and identified). We repeated
the sample level aggregation of taxa on the 300-count file; thus, the resultant site-level count
file typically had a total count of 600 individuals. We did not calculate density on the 300-count
files but did calculate biomass.
7.4 Zooplankton MMI development
7.4.1 Regionalization
We divided the conterminous US into five "bio-regions" based on nine aggregated Omernik
Level III ecoregions (Omernik 1987, Stoddard 2004, Herlihy et al. 2008, Omernik and Griffith
2014) that were developed for use on NARS reporting Figure 7.1). We combined the Northern
and Southern Appalachian regions (NAP, SAP) into a single bio region (Eastern Highlands,
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EHIGH). We combined the three "plains" regions (Northern, Southern, and Temperate [NPL,
SPL, and TPL]) into a single bio-region (PLAINS), In the western US, we combined the Xeric and
Western Mountains regions (XER, WMT) into a single "Western Mountains" bio-region
(WMTNS). Despite relatively small sample sizes of least disturbed sites, we kept the Coastal
Plains (CPL) and Upper Midwest (UMW) as separate bio-regions. These are the same regions as
are used for the NLA benthic macroinvertebrate MMI.
Figure 7.1 Five aggregated bio-regions used to develop zooplankton MMIs for the 2012 National Lake Assessment
(CPL=Coastal Plains; EHIGH=Eastern Highlands, PLAINS= Plains, UMW=Upper Midwest, and WMTNS=Western
Mountains). Solid dots indicate least disturbed sites used for developing the zooplankton MMI. White circles
indicate least disturbed sites that we excluded because of atypical samples (too few taxa or number of individuals
collected).
7.4.2 Least and most disturbed sites
For NLA 2012, we used the same list of sites for the zooplankton MMI as those selected for
benthic macroinvertebrates (RT_NLA12; see Section 3.3). We identified two least disturbed
sites that appeared to have abnormal zooplankton samples and excluded them from the list of
least-disturbed sites.
For NLA 2017, we combined least disturbed sites sampled in 2017 with those from the NLA
2012 assessment. We retained only one visit per site by excluding revisits (VISIT_NO=2) and
using the 2017 visit for sites from 2012 that were resampled in 2017. In addition, we identified
three situations where we felt that the zooplankton samples from least disturbed sites were
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not representative of the existing assemblage, and excluded these sites from developing
condition class benchmarks:
1. Sites where at least one of the net samples were hugely dominated by unidentified
copepod individuals (which included nauplii and immature copepodites).
2. Sites where no rotifers were collected.
3. Sites where less than 100 individuals were collected in either the coarse or fine net
sample.
The first two of these situations were used in the NLA 2012 assessment. The third situation was
added for the NLA 2017 assessment. Out of 343 least disturbed lakes, we identified 21 sites (15
from 2012 and only six from 2022) where the coarse sample had less than 100 individuals
counted. At two of these sites (both from 2012), the fine net sample also had less than 100
individuals counted.
For NLA 2022, we identified least-disturbed and most disturbed sites using the same process as
described above and in Section 3.3. However, we did not apply the additional screening criteria
specific to zooplankton samples and did not use sites from 2022 to modify the existing
condition class benchmarks.
7.4.3 Least disturbed sites: calibration versus validation
As an independent check on the MMI developed for each bio-region, we set aside a small
number of least disturbed sites as "validation" and did not include them in any MMI or metric
evaluations or performance testing. We used revisit sites (typically VISIT_NO=2) as validation
sites because they are not used in any metric or MMI testing. We then supplemented the list of
revisit sites in each region by randomly selecting sites from the list of least disturbed sites.
Where possible, we withheld ~10% of the least disturbed sites in each bio-region as validation
sites, leaving at least 15 least disturbed sites available for developing and evaluating metrics
and MMIs. For the CPL and UMW bio-regions, the small number of least disturbed sites
prevented setting aside 10% of the site for validation. Numbers of validation sites were as
follows: CPL (8), EHIGH (16), PLAINS (14), UMW (10), and WMTNS (18).
7.4.4 Candidate metrics
We used the count data file and the master taxa list file to calculate candidate metrics. We
assigned candidate metrics to one of six metric categories, with each category reflecting a
different attribute of assemblage structure or ecological function.
The Abundance category included metrics based on abundance, density, or biomass. We
calculated these metrics separately for the ZOFN samples, the ZOCN samples, and for the
combined samples. Within the combined sample, we also calculated abundance metrics
separately for the net-based size classes (COARSE vs. FINE).
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The Richness category included metrics based on taxa richness and metrics related to taxa
diversity or dominance. Richness metrics included total distinct taxa richness, number of
genera, and number of families. We calculated these metrics separately for the ZOCN, ZOFN,
and combined sample. We calculated diversity and dominance metrics for the combined
sample based on abundance, density, and biomass. Diversity metrics included Shannon-Weiner
and Simpson indices, and Hurlbert's Probability of Interspecific Encounter (PIE, Hurlbert 1971,
Jeppesen et al. 2000). For each combined net sample, we developed dominance metrics based
on the percent of individuals represented in the most dominant taxon and represented in the
three and five most dominant taxa.
We assigned separate categories for each of the three principal taxonomic components of the
zooplankton assemblage: Cladoceran, Copepod, and Rotifer. Metrics in these three categories
included abundance and richness metrics calculated separately for each taxonomic group. For
copepods, we also calculate the ratio of calanoids to the sum of cladocerans and cyclopoids,
following Gannon and Stemberger (1978) and Kane et al. (2009).
The sixth metric category was trophic guild. We identified three major guilds, herbivores,
omnivores, and predators. Each taxon was assigned to a trophic guild based on information
from the Northeast Lakes Survey (Stemberger and Lazorchak 1994, Stemberger et al. 2001). We
calculated metrics using both the entire sample and for the 300-count rarefied samples. Metrics
derived from the rarefied sample have "300" in the variable name.
For many metrics, we could calculate six different variants: the number of distinct taxa
(metr/'c_NTAX), total biomass (metric_BIO), density (metr/'c_DEN), percent of individuals
(metr/'c_PIND), percent of total biomass (metric_PB\0) and percent of total density
(metr/'c_PDEN). We did not calculate density-based metrics for the 300-count rarefied samples.
Each variant was calculated based using all the individuals in the sample, and for just the native
individuals in the sample. We calculated a total of 374 candidate metrics for the whole sample
count data, and an additional 272 metrics from the 300-count rarefied sample data.
7.4.5 Final metric selection
We subjected all of the candidate metrics to five screening procedures, following Stoddard et
al. (2008). The first was a range test. We excluded richness metrics (metr/'c_NTAX) with a range
of <4 from further consideration. We excluded metrics based on biomass (metric_BIO), density
(metr/'c_DEN), diversity metrics, and zooplankton ratio if the 90th percentile (P90) was 0. We
excluded percentage metrics (metric_PTAX, metr/'c_PBIO, metr/'c_PDEN) if the 75th percentile
(P75) was <10%.
The second screen was a signal to noise (S:N) test, following Kaufmann et al. (1999). We
compared the total variance observed across all sites (signal) against the variance observed for
sites that were sampled twice in the same index period (noise). We excluded metrics that had
S:N values < 1.25.
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The third screen was for responsiveness to disturbance. For each metric, we calculated the t-
statistic for each metric comparing values for the set of least disturbed sites with those for the
set of most disturbed sites. We considered metrics having 11\ values < 1.73 as non-responsive
to disturbance.
The fourth screen was to determine if metrics required adjustment for lake size. We generated
plots of linear regressions of each metric with lake area (AREA_HA) to determine if the metric
response changed with increasing lake size. For all metrics, the upper 95% prediction interval at
the minimum response value overlapped the lower 95% prediction interval at the maximum
response value, indicating there was no significant effect of lake size on the metric response.
For each bio-region, we used the set of candidate metrics that had passed the four screens
describe above to develop candidate MMIs. We constrained the MMIs to contain at least one
metric from each of the six metric categories (abundance, richness, crustacean, copepod,
rotifer, and trophic). If no metrics within a category passed all of the screens, we selected one
or more metrics that had the highest tvalues and had S:N values near 1 (if possible). Values of
S:N <1 indicate that that variation within a site is equal to or greater than the variation among
sites, so the metric cannot discriminate among sites.
Finally, we evaluated the redundancy among candidate metrics using correlation analysis.
Historically, we have evaluated redundancy based on the establishing a maximum allowable
correlation coefficient (r) between two metrics (e.g., r >0.7; Stoddard et al. 2008)). Van Sickle
(2010) demonstrated that MMIs containing a suite of metrics that have a low average
correlation among them perform better that simply using a maximum threshold value of r to
reduce redundancy within the suite of metrics. We included correlations in the procedure
below, computing correlations among metrics for each candidate MMI, rather that evaluating
individual input metrics within a category and choosing only non-redundant metrics to include
in a final MMI, as described by Stoddard et al. (2008).
Candidate metrics that we considered for inclusion into an MMI for each of the five bio-regions
are listed in Appendix D: Zooplankton. For each bio-region, we computed MMIs from all
possible combinations of candidate metrics from the six categories. We evaluated each MMI for
responsiveness (t test of least disturbed vs. most disturbed sites) and repeatability (S:N). For
each bio-region, we selected MMI that had a combination of high t value, a reasonable value
for S:N, low mean r among the suite of metrics, and, when possible, a maximum value of r for
the suite of metrics that was <0.7.
NOTE: As described in Section 7.2.1, we had to recalculate metrics and MMI scores after
correcting for the error in the calculation of tow volume. We repeated the metric screening
process and determined that the existing suite of metrics included in each of the regional MMIs
still performed adequately, so we retained them for use in the NLA 2022 assessment. The
results presented in Section 7.5 are based on the recalculated data.
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7.4.6 Metric scoring
We followed the approach described by Stoddard et al. (2008) to transform metric responses
into a metric score that ranged between 0 and 10 (Blocksom 2003). For positive metrics (i.e., t
>0), we used the 5th percentile of all sites in the bio-region as the "floor" value, and the 95th
percentile of the set of least disturbed sites as the "ceiling" value. For negative metrics (i.e., t
<0), we used the 5th percentile of least disturbed sites in the bio-region as the "floor" value, and
the 95th percentile of all sites as the "ceiling" value. When metric response values were less
than the floor value, we assigned a score of 0. When metric response values were greater than
the ceiling, we assigned a score of 10. We estimated scores for response values that were
between the floor and ceiling values by linear interpolation.
We calculated the final MMI score for each bio-region by summing the six component metric
scores, and then multiplying by 10/6. This resulted in an MMI score that ranged between 0 and
100.
7.5 Zooplankton MMI metric composition and performance
See Appendix D: List of Candidate Metrics for Zooplankton for metric descriptions.
7.5.1 Coastal Plains MMI
The component metrics for the Coastal Plains MMI are presented in Table 7-2. Information
related to the performance of the Coastal Plains MMI are presented in section 7.6. Figure 7.2.
compares the distributions of the six metrics in least disturbed vs. most disturbed sites. Three
metrics are "negative" metrics (t <0) values, indicating that the response is greater in most
disturbed sites compared to least disturbed sites. No abundance or cladoceran metrics passed
both the responsiveness and repeatability screens. The abundance metric (FINE_BIO [biomass
of smaller-sized taxa]) had a t value and an S:N value that were just below the screening
criterion. The cladoceran metric (SIDlD_PIND [percent of individuals of the cladoceran family
Sididae]) had an S:N value that was below the screening criterion.
The abundance metric (FINE_BIO), the cladoceran metric (SIDlD_PIND), the richness metric
(FAM300_NAT_NTAX), and the trophic metric (OMNI_PTAX) responded as expected to
disturbance as expected (Figure 7.2; Table 7-1). The copepod metric (DOM1_300_COPE_PBIO)
and the rotifer metric (COLLO_PBIO) decreased in response to disturbance (Figure 7-2).
Declines in the proportion of total biomass contributed by either dominant copepods or a
subgroup of rotifers might be expected if the total richness and abundance total biomass of
cyclopoid copepods and rotifers increased with disturbance (Table 7-1).
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Table 7-2. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE COASTAL PLAINS BIO-REGION.
Evaluations for responsiveness (t-value) and signahnoise (S:N) based on index visits and do not include least
disturbed "validation" sites. Negative values for t indicate response is greater in most disturbed sites vs. least
disturbed sites. Metrics having values marked with an asterisk were among the best performing metric of that
category but failed one or more evaluation screens. Floor and ceiling values are used to derive a score for the
metric.
Metric Type
Metric Variable Name (floor, ceiling)
t value
S:N (bio-region)
Abundance/Size
FINE_BIO (2.913623, 173.279784)
-1.67*
1.2*
Cladoceran
SID!D_PIND (0, 24.88)
-1,80
0.4*
Copepod
DOM1_3DO_COPE_PBIO (45.90, 100)
+1.16*
1.9
Richness/Diversity
FAM300_NAT_NTAX (5, 15)
+2.72
2.1
Rotifer
COLLO_PBIO (0, 5.64)
+1.84
7.2
Trophic
OMNLPTAX (10.53, 47.06)
-3.35
4.3
ABUNDANCE
CLADOCERAN
COPEPOD
o
m
RICHNESS
ROTIFER
TROPHIC
60
50
5 40
|j 30
z
O 20
10
0
•
T
V
_L
•
Figure 7.2. Distribution of six component metrics of the zooplankton MMI for the Coastal Plains bio-region in least
disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th percentiles.
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7.5.2 Eastern Highlands MMI
The component metrics for the Eastern Highlands MMI are presented in Table 7-3. Information
related to the performance of the Eastern Highlands MMI are presented in section 7.6. Figure
7.3 compares the distributions of the six metrics in least disturbed vs. most disturbed sites. The
suite of metrics includes both positive (2) and negative (4) metrics. No richness metrics passed
the screens for responsiveness or repeatability. The richness metric (ZOCN300_FAM_NTAX) had
a t value (1.64) just below the screening criterion, while the S;N value (0,3) was well below the
screening criterion.
The cladoceran metric (SMCLAD_PBIO), the richness metric (COARSE_NAT_PTAX ),the rotifer
metric (ROT_PBIO), and the trophic metric (OMNI300_PTAX) responded as expected to
increased disturbance (Figure 7.3; Table 7-1). The abundance metric (ZOCN_DEN) and the
copepod metric (COPE_NAT_DEN) both increased in response to disturbance (Error! Reference
source not found.)- An increase in cyclopoid copepods expected with increased disturbance
(Table 7-1) would help to explain the observed response in both of these metrics.
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Table 7-3. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE EASTERN HIGHLAND BIO-REGION.
Evaluations for responsiveness (t-value) and signahnoise (S:N) based on index visits and do not include least
disturbed "validation" sites. Negative values for t indicate response is greater in most disturbed sites vs. least
disturbed sites. Floor and ceiling values are used to derive a score for the metric. See Appendix D: Zooplanktonfor
metric descriptions.
Metric Type
Metric Variable Name (floor, ceiling)
t value
S:N (bio-region)
Abundance/Size
ZOCN_DEN (0.216450,259.3050)
-1.89
2.2
Cladoceran
SMCLAD_PBIO (0, 57.31)
-2.91
1.3
Copepod
COPE_NAT_DEN (8,8236,398.397)
-1.70
1.5
Richness/Diversity
COARSE_NAT_PTAX (22.22,57.14)
+1.71*
0.2*
Rotifer
ROT_PBIO (0.79,86.39)
-1.94
1.2*
Trophic
OMNI300_PTAX (12.50, 44.44)
-2.48
1.8
ABUNDANCE
CLADOCERAN
COPEPOD
z
UJ
a
o
o
N
m
0- 60
Q
<
d 40
s
tn
T
i=L
100
7
80
LU
O
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60
<
Z
UJ1
40
n
O
o
20
0
RICHNESS
ROTIFER
TROPHIC
60
§ 50
|j 40
<
Z, 30
LU
CO -A
a: 20
8 10
X
T
100
80
O
CO 60
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O 40
a
20
0
60
50
J?
1-
40
0.
o
30
CO
Z
5
20
O
10
0
JL
Figure 7.3 Distribution of six component metrics of the zooplankton MMI for the Eastern Highlands bio-region in least
disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th percentiles.
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7.5.3 Plains MM!
The component metrics for the Plains MMI are presented in Table 7-4. Information related to
the performance of the Plains MMI are presented in section 7.6. Figure 7.4 compares the
distributions of the six metrics in least disturbed vs. most disturbed sites. The MMI was
comprised of two negative and four positive metrics. All metrics passed the screening criteria
for both responsiveness and repeatability.
The copepod (COPE_RATIO_300_BIO), richness (FAM300_NAT_TAX), and the trophic
(COPE_HERB_PDEN) metrics responded as expected to increased disturbance (Figure 7.4; Table
7-1). The abundance (FINE300_NAT_PBIO), cladoceran (SMCLAD_NAT_PIND), and the rotifer
(ROT_NTAX) metrics all decreased with response to increased disturbance. If herbivorous
cyclopoid copepods are becoming more dominant in terms of richness, abundance, and
biomass, that may result in a decline in the relative biomass of individuals collected in the fine-
mesh net (principally rotifers), a decline in the relative abundance of smaller cladocerans, and a
decline in rotifer taxa richness.
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Table 7-4, COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE PLAINS BIO-REGION,
Evaluations for responsiveness (t-value) and signahnoise (S:N) based on index visits and do not include least
disturbed "validation" sites. Negative values for t indicate response is greater in most disturbed sites vs. least
disturbed sites. Floor and ceiling values are used to derive a score for the metric.
Metric Type
Metric Variable Name (floor, ceiling)
t value
S:N (bio-region)
Abundance/Size
FINE300_NAT_PBIQ (0.66, 85.12)
+1,89
5.8
Cladoceran
SMCLAD_NAT_PIND (0, 49.03)
+3.11
1.8
Copepod
COPE_RATIQ_300_BIO (0, 62.81)
+2,41
3.0
Richness/Diversity
FAM300_NAT_NTAX (5, 15)
+2,20
2.6
Rotifer
ROT_NTAX (3, 17)
+2.63
1.7
Trophic
COPE_HERB_PDEN (0, 29.93)
-2.45
9.1
ABUNDANCE
CLADOCERAN
COPEPOD
o
m
a.
60 -
o
Z
50 -
•
-
a
I
40
•
<
z
30 -
--
a
<
_i
o
20
:
t/>
10 -
0 J
—•—
—w—
RICHNESS
ROTIFER
TROPHIC
16
12
i 4
L M
Figure 7.4. Distribution of six component metrics of the zooplankton MMI for the Plains bio-region in least
disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th percentiles.
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7.5.4 Upper Midwest MMI
The component metrics for the Upper Midwest MMI are presented in Table 7-5. Information
related to the performance of the Upper Midwest MMI are presented in section 7.6. Figure 7.5
compares the distributions of the six metrics in least disturbed vs. most disturbed sites. The
MMI is composed of four negative and two positive metrics. No abundance metrics passed the
screen for responsiveness. The abundance metric (ZOCN_NAT_PDEN [the percent of total
density represented by native individuals in the coarse net sample]) had a t-value that is below
the screening criteria for responsiveness. Repeatability (S:N values) of the metrics in this bio-
region are higher than in other bio-regions, but interpretation of the S:N values is constrained
somewhat by a limited number of revisit samples (5). The value for the abundance metric
(2348) resulted from essentially no difference in the values between the small number of revisit
samples.
Only three of the six metrics responded to disturbance as expected (Figure 7.5Error! Reference
source not found.; Table 7-1). The abundance metric (TOTL_NAT_PIND) showed a slight
decrease with disturbance, indicating the effect of non-native taxa in this bio-region. The rotifer
metric (DOMl_ROT_PBIO) indicates a reduction in species richness (i.e., increased dominance
by one or a few taxa) with increased disturbance. The trophic metric (COPE_HERB300_PBIO)
indicates an increase in herbivorous taxa (possibly cyclopoid copepods) with increased
disturbance. The cladoceran metric (BOSM300_NAT_PTAX) was expected to increase with
increased disturbance, but the response may reflect a larger increase in the taxa richness of
other forms of smaller zooplankton (e.g., cyclopoid copepods). The copepod metric
(CALAN300_NAT_BIO) indicates an increase in larger forms of zooplankton. Such a response
might occur if the least disturbed population of lakes is dominated by oligotrophic lakes that do
not support large populations of zooplankton. The richness metric (FINE_PTAX) decreased in
response to disturbance. This response may be similar to that observed for the cladoceran
metric, where other forms of smaller zooplankton (e.g., cyclopoid copepods) increase in
taxonomic richness compared to rotifers, which are the dominant taxa collected in the fine-
mesh net.
7.5.5 Western Mountains MMI
The component metrics for the Western Mountains MMI are presented in Table 7-6.
Information related to the performance of the Western mountains MMI are presented in
Section 7.6. Figure 7.6 compares the distributions of the six metrics in least disturbed vs. most
disturbed sites. The MMI is composed of three negative and three positive metrics. No richness
metrics passed the screen for responsiveness. The richness metric (ZOFN300_NTAX [Number of
distinct taxa in the 300-count rarefied sample from the fine net sample]) had a t value that was
below our acceptance criteria for responsiveness.
The abundance (COARSE300_NAT_PBIO), cladoceran (LGCLAD300_NAT_PTAX), richness
(ZOFN300_NTAX), rotifer (PLOIMA_PTAX), and trophic (COPE_OMNI_PTAX) metrics responded
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as expected to increased disturbance (Figure 7.6, Table 7-1). The copepod metric
(COPE300_BIO) would respond as expected to disturbance if the increase in biomass was due
primarily to smaller forms (e.g., cyclopoid copepods).
Table 7-5. COMPONENT METRiCS OF THE ZOOPLANKTON MMi FOR THE UPPER MiDWEST BIO-REGION.
Evaluations for responsiveness (t-value) and signahnoise (S:N) based on index visits and do not include least
disturbed "validation" sites. Negative values for t indicate response is greater in most disturbed sites vs. least
disturbed sites. Metrics having values marked with an asterisk were the best performing metric of that category
but failed one or more evaluation screens. Floor and ceiling values are used to derive a score for the metric.
Metric Type
Metric Variable Name (floor, ceiling)
t value
S:N (bio-region)
Abundance/Size
TOTL_NAT_PIND (96.75, 100)
+1,47*
2348
Cladoceran
BOSM300_NAT_PTAX (0, 12.5)
+2.72
1.3
Copepod
CALAN300_NAT_BIO (0,65.037544)
-2.17
9.9
Richness/Diversity
FINE_PTAX (37.50, 77.78
+1.87
1.4
Rotifer
DOMl_ROT_PBIO (25.30, 93.60)
-2.46
3.5
Trophic
COPE_HERB300_PBIO (0.19, 59.42)
-1.99
5.1
ABUNDANCE
CLADOCERAN
100
£L
5
20
15
10
8 5
CD
T_
RICHNESS
ROTIFER
TROPHIC
100
80
60 -
40
20 -
0
O
m
a.
h"
0
ZL
I
1
o
Q
80
O
to
Q- 60
I
o
0
CO
£ 40
Lli
1
£ 20
o
O
I
T-
L M
Figure 7.5. Distribution of six component metrics of the zooplankton MMI for the Upper Midwest bio-region in
least disturbed (L) versus most disturbed (M) sites. Dots indicate the 5th and 95th percentiles.
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Table 7-6. COMPONENT METRICS OF THE ZOOPLANKTON MMI FOR THE WESTERN MOUNTAINS BIO-REGION.
Evaluations for responsiveness (t-value) and signahnoise (S:N) based on index visits and do not include least
disturbed "validation" sites. Negative values for t indicate response is greater in most disturbed sites vs. least
disturbed sites. Metrics having values marked with an asterisk were the best performing metric of that category
but failed one or more evaluation screens. Floor and ceiling values are used to derive a score for the metric.
Metric Type
Metric Variable Name (floor, ceiling)
t value
S:N (bio-region)
Abundance/Size
COARSE30Q_NAT_PBIO (10.94, 99.26)
+1,89
5.6
Cladoceran
LGCLAD300_NAT_PTAX (0, 29.285)
+2.53
2.0
Copepod
COPE300_BIO (0.073928, 149.035677)
-2,75
2.0
Richness/Diversity
ZOFN300_NTAX (3, 15)
-1.69*
1.9
Rotifer
PLOIMA_PTAX (20, 70.835)
+0.49*
4.3
Trophic
COPE_OMNI_PTAX (0, 22.22)
-2.46
1.5
o
DO
Q-
Ul
cn
a:
<
o
o
ABUNDANCE
CLADOCERAN
COPEPOD
i-
o.
<
Q
<
O
O
400
300 -
200 -
LU
Q.
O
O
100
RICHNESS
ROTIFER
TROPHIC
20
g 15
i-
z
g' 10
O c
N 5
X
<
<
s
o
0.
30
O., 20
a' 1"
o
o
T
X"
Figure 7.6. Distribution of six component metrics of the zooplankton MMI for the Western Mountains bio-region in
least (L) disturbed versus most disturbed (M) sites. Dots indicate the 5th and 95th percentiles.
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7.6 Zooplankton MMI performance
We evaluated each of the five regional MMIs in several ways.
7.6.1 Calibration versus validation sites
To provide an independent assessment of MMI performance, we compared the distribution of
MMI scores between the set of validation sites (which we did not use in MMI development) and
the calibration sites using a t-test. The null hypothesis was that the mean values of the two
groups would be equal. Mean values of the two groups were not significantly different (p <
0.05) for any bio-region (Table 7-7). Error! Reference source not found, shows the distribution
of MMI scores between the calibration and validation sites in the five bio-regions.
7.6.2 Precision of MMIs based on least disturbed sites
We evaluated the precision of the regional MMIs using the sets of least disturbed calibration
sites, following Van Sickle (2010). We rescaled the MMI scores in each bio-region by dividing
each site score by the mean MMI score, which resulted in a mean rescaled MMI score of 1. We
calculated the standard deviation of the rescaled MMI scores (Table 7-7). The smaller the
standard deviation, the more precise the index is, and the better the ability to detect sites that
are not in least disturbed condition. Standard deviations were generally small except for the
Plains, where site MT-104 had a large influence.
7.6.3 Responsiveness, redundancy, and repeatability of zooplankton MMIs
We compared the MMI scores from the set of least disturbed sites to the set of most disturbed
sites (excluding the validation sites) using a t-test. We calculated the S:N values using the set of
revisit sites within each bio-region (again excluding the validation sites). Table 7-8 presents the
results of these tests, along with the maximum and average correlations observed for the
component metrics. The t values for responsiveness are comparable to MMIs developed for
other resource types and assemblages (e.g., benthic invertebrates)Figure 7.8 Distribution of
zooplankton MMI scores in least-disturbed (L) vs. most disturbed (M) sites for five bio-regions.
Sample sizes are in parentheses. Dots indicate the 5th and 95th percentiles. Figure 7.8Error!
Reference source not found, shows the distribution of MMI scores between least- and most
disturbed sites in the five bio-regions. SignakNoise values are comparable to other MMIs that
have been developed for other assemblages. The S:N value for the UMW bio-region is
constrained by the small number of revisit sites (5) available. When MMI scores from all bio-
regions are considered, the national-level estimate of S:N is 7.0.
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Table 7-7. RESULTS OF INDEPENDENT ASSESSMENT AND PRECISION TESTS OF NLA 2012 ZOOPLANKTON MMIs
BASED ON LEAST DISTURBED SITES,
None of the t-values were significant at p = 0.05. Standard deviations were calculated using only calibration sites.
Calibration vs. Validation
Standard Deviation
Sites
of Standardized
Regional MMI
(t-value)
MMI scores
Coastal Plains (CPL)
0.85
0.187
Eastern Highlands (EHIGH)
-1.23
0.119
Plains (PLAINS)
1.21
0.237
Upper Midwest (UMW)
0.94
0.112
Western Mountains (WMTNS)
0.42
0.117
100
VN* ^ ^ # \N ^ ^
//////////
G^^V V^V V^V V^V
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NLA 2022 Technical Support Document-August 2024
Table 7-8. RESULTS OF RESPONSIVENESS, REDUNDANCY, AND REPEATABILITY TESTS FOR NLA 2012 ZOOPLANKTON
MMIs.
* For the Upper Midwest (UMW) MMI, the abundance metric scores in all least-disturbed sites were identical.
Values in parentheses are correlation coefficients with the abundance metric coefficients set to missing.
Redundancy
Responsiveness
Redundancy
(Mean pairwise
Repeatability
t-test of Least
(Maximum pairwise
correlation among
Signal: Noise ratio
disturbed vs. Most
correlation among
component
based on revisit
Bio-Region
disturbed Sites
component metrics)
metrics)
sites
Coastal Plains
(CPL)
4.11
0.55
0.28
2.7
Eastern Highlands
(EHIGH)
5.09
0.43
0.17
2.5
Plains (PLAINS)
5.49
0.57
0.20
3.6
Upper Midwest
(UMW)
5.78
1.0 (0.61)*
0.50 (0.20)*
18.0
Western
Mountains
(WMTNS)
6.28
0.561
0.20
3.6
100
LU
t£
O
o
CO
o
I—
*
z
<
_l
Q.
O
o
N
Least Disturbed (L) vs. Most Disturbed (M)
(Index visits only. For L: Calibration sites only)
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V > V1 . v v vV , V w
/ v ' \X .VN .C-i 6.'n .a A'n .c*> fi/1
«,v // *
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NLA 2022 Technical Support Document - August 2024
7.6.4 Responsiveness to a generalized stressor gradient
We performed an additional evaluation of the MMIs for responsiveness to disturbance. We
performed principal components analysis (PCA) on the set of chemical, physical habitat, and
visual assessment stressor variables used to screen for least disturbed and most disturbed sites.
Chemical stressor variables included chloride, sulfate, turbidity, and acid neutralizing capacity
(CL, S04, TURB, and ANC, respectively). Habitat stressor variables (Kaufmann et al. 2014; see
Chapter 5 for descriptions and calculations) included shoreline disturbance due to non-
agricultural activities (hiiNonAg), shoreline disturbance due to agricultural activities (hiiAg_Syn),
and the proportion of shoreline stations with at least one type of disturbance present in either
the littoral zone or shoreline plots (hifpAnyCirca_syn). Stressor variables from the visual
assessment included the intensity of observed types of agricultural activities (AGR_SCORE),
intensity of observed types of residential activities (RES_SCORE), and intensity of observed
types of commercial and industrial activities, excluding evidence of fire (IND_NOFIRE). We
transformed the chemical variables (logio[x+l]) and standardized all variables to mean=0 and
variance=l. The first PCA axis explained 38% of the total variance, and the highest variable
loadings were for the chemical and agricultural-related habitat variables. The second PCA axis
explained an additional 18% of the total variance, and the highest variable loadings were for
the non- agricultural habitat variables and the intensity of residential activities. Linear
regression of the MMI score versus the PCA axis 1 scores yielded an r2 of 0.42 (r= 0.65) for PCA
axis 1 (Figure 7-9), and 0.006 for PCA axis 2 scores. These results indicate the zooplankton MMI
is principally responsive to nutrient conditions resulting from agricultural disturbance, and less
responsive to other types of habitat disturbance.
7.6.5 Effect of natural drivers and tow length on MMI scores
The set of lakes sampled for the NLA 2012 included both natural and human-made lakes and
included a wide range of sizes (as estimated by lake area as represented in NHD). In addition,
the sampling protocol did not include a vertical tow through the entire water column. Any one
of these factors might produce a bias in the MMI scores that would require assessing ecological
condition separately for one or more of these groups of lakes (natural vs. human-made, small
vs. large lakes, or shallow versus deeper lakes). We use the set of least disturbed sites
(calibration and validation) to evaluate the potential differences in MMI scores in these groups
of lakes.
7.6.5.1 Lake origin
We compared the distributions of MMI scores in least disturbed natural lakes vs. human-made
reservoirs for each of the five bio-regions (Figure 7.10). The distributions are similar within each
bio-region except the WMTNS, where human-made lakes appear to have much lower MMI
scores than natural lakes. In the Coastal Plains, human-made lakes have higher MMI values than
natural lakes, but interpretation is constrained by the small number of least disturbed natural
lakes (n=3). In the WMTNS, the sample size for least disturbed human-made lakes is relatively
small (n=16) and is influenced to some extent by the presence of outliers with low MMI scores
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(Figure 7.10). We did riot feel the observed differences were large enough to treat MMI scores
from lakes and reservoirs differently in terms of settings condition benchmarks.
MMI= -11.94(PCA Axisl Score) + 55.97 (R2=0.42)
100
-4 -2 0 2 4 6
PCA Axis 1 Score
Figure 7.9. Linear regression of NLA 2012 Zooplankton MMI scores vs. first axis score from principal components
analysis (PCA) based on chemical, habitat, and visual assessment stressor variables used to screen least- and most
disturbed sites.
Least Disturbed Sites
c
o
c
ro
a.
o
o
N
100
80
60
40
20
0
S
~
i
MAN-MADE
NATURAL
5
i
-1-
T
1
_L
CPL EHIGH PLAINS UMW WMTNS
(20,3) (26,35) (23,16) (0,31) (17,43)
Bio-region
Figure 7.10. NLA 2012 Zooplankton MMI scores of human-made (shaded boxes) versus natural lakes (unshaded
boxes) for least disturbed sites in five bio-regions. See Figure 7.1 for bio-region codes. Sample sizes for each type
are in parentheses. Dots indicate 5th and 95th percentiles.
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7.6.5.2 Lake size
We examined the set of least disturbed sites for evidence of difference in MMI scores due to
lake size (Figure 7.11). We noted earlier than we did not have to calibrate individual metrics for
lake size (Section 7.4.5). Distributions of MMI scores were similar in median values and ranges
for all size classes except for the largest (> 500 ha), which had a similar median but a wider
range.
Least Disturbed Sites
o
i.
o
o
(/>
c
o
100
80
60
3 40
c
m
Q.
©
©
N
20
V?
<§>
$
\N
N*
V?
4f
Figure 7.11. Zooplankton MMI scores versus lake size class within least disturbed lakes of the NLA 2012. Sample
sizes are in parentheses. Dashed lines are mean values. Dots indicate the 5th and 95th percentiles.
7.6.5.3 Site depth
We had some concerns that the 5-m tow length used to collect zooplankton samples might be
less effective in deeper lakes, where larger taxa may migrate to deeper waters during the day
to avoid fish predation, and thus be underrepresented in the samples. We examined MMI
scores in least disturbed sites as they related to the depth of the index site where samples were
collected (Figure 7-12). There was no apparent pattern in relation to site depth, and the
distribution of MMI scores was similar for least-disturbed lakes that were < 6 m deep (the
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maximum depth where the tow length encompassed the entire water column), and for lakes >
6 m deep (where part of the water column would not be subject to sampling).
7.6.5.4 Component metrics used in zooplankton MMIs for NLA 2022
Table 7-9 summarizes the component metrics for each of the five zooplankton MMIs used in NLA
2022. There were no changes or modifications from those used in the NLA 2012.
Table 7-9. Component metrics of the zooplankton multimetric indices (MMIs) used for NLA 2022.
Metric Category
Metric Description
Direction
of
Response"
Metric Variable Name
Coastal Plains MMI
Abundance/Biomass/Density
Biomass of fine mesh net (50 nm) taxa
INC
FIN E_BIO
Cladoceran
% of total individuals that are within
the cladoceran family Sididae
INC
SIDID_PIND
Copepod
% of biomass in dominant copepod
taxon (300 count subsamples)
DEC
DOM1_300_COPE_BIO
Richness/Diversity
Number of native families (300 count
subsamples)
DEC
FAM300_NAT_NTAX
Rotifer
% of total biomass within the rotifer
order Collothecaceae
DEC
COLLO_PBIO
Trophic
% of taxa that are omnivorous
INC
OMNI_PTAX
Eastern Highlands MMI
Abundance/Biomass/Density
Density of individuals collected in
coarse mesh net (150-nm)
INC
ZOCN_DEN
Cladoceran
% Biomass represented by small
cladoceran individuals
INC
SMCLAD_PBIO
Copepod
Density represented by native copepod
individuals
INC
COPE_NAT_DEN
Richness/Diversity
% of taxa that are larger-sized and
native
DEC
COARSE_NAT_PTAX
Rotifer
Percent total biomass from rotifers
INC
ROT_PBIO
Trophic
Percent of taxa that are omnivorous
(300-count subsamples)
INC
OMNI300_PTAX
Plains MMI
Abundance/Biomass/Density
% of biomass represented in individuals
of smaller-sized native taxa (300-count
subsamples)
DEC
FINE300_NAT_PBIO
Cladoceran
% of native individuals within the
suborder Cladocera that are "small"
(coarse and fine net samples
combined)
DEC
SMCLAD_NAT_PIND
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Metric Category
Metric Description
Direction
of
Response"
Metric Variable Name
Copepod
Ratio of Calanoids to
(Cladocera+Cyclopoids) based on
biomass (300-count subsamples).
DEC
COPE_RATIO_300_BIO
Richness/Diversity
Total native family richness (300-count
subsamples)
DEC
FAM300_NAT_NTAX
Rotifer
Number of rotifer taxa
DEC
ROT_NTAX
Trophic
% of total density represented by
herbivorous copepods
INC
COPE_HERB_PDEN
Upper Midwest MMI
Abundance/Biomass/Density
% of native individuals
DEC
TOTL_NAT_PIND
Cladoceran
% of native taxa that are within the
cladoceran family Bosminidae (300-
count subsamples)
DEC
BOSM300_NAT_PTAX
Copepod
Biomass of individuals within native
calanoid taxa (300-count subsamples)
INC
CALAN300_NAT_BIO
Richness/Diversity
% of fine mesh net (50 nm) taxa
DEC
FINE_PTAX
Rotifer
Percent of rotifer biomass in dominant
rotifer taxon
INC
DOMl_ROT_PBIO
Trophic
Percent of biomass represented by
herbivorous copepods (300-count
subsamples)
INC
COPE_HERB300_PBIO
Western Mountains MMI
Abundance/Biomass/Density
% biomass of individuals of native
coarse mesh net (150 nm) taxa (300-
count subsamples)
IDEC
COARSE300_NAT_PBIO
Cladoceran
% of distinct native taxa that are large
cladocerans (300-count subsamples)
DEC
LGCLAD300_NAT_PTAX
Copepod
Total biomass of copepod individuals
within the subclass Copepoda (300-
count subsamples)
INC
COPE300_BIO
Richness/Diversity
Number of taxa in the fine net (50-nm)
sample (300-count subsample)
INC
ZOFN300_NTAX
Rotifer
% taxa that are within the rotifer order
Ploima
DEC
PLOIMA_PTAX
Trophic
% taxa that are omnivorous copepods
INC
COPE_OMNI_PTAX
0 Direction of response to increased disturbance: INC= response increases with increased disturbance,
DEC=response decreases with increased disturbance.
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7.7 Thresholds for assigning ecological condition
7.7.1 NLA 2012
For the NLA 2012, we followed Stoddard et al. (2008) in using the set of least disturbed sites
(including calibration and validation sites) to set ecological condition benchmarks based on the
zooplankton MMI. We used the 25th percentile value to distinguish sites in "good" condition (similar
to least disturbed) from sites in "fair" condition (slightly deviant from least disturbed). We used the
5th percentile value to distinguish sites in "fair" condition from sites in "poor" condition (different
from least disturbed).
Because of varying quality of least disturbed sites within each bio-region, we adjusted the
percentiles using the same process as for the NLA 2012 benthic macroinvertebrate indicator
(Herlihy et al. 2008; see Chapter 6). We performed principal components analysis (PCA) based
on all variables used in the screening of least disturbed sites (TP, TN, CI, S04, Turbidity, physical
habitat disturbance indices, and assessment indices). We transformed values (logio[x] or
logio[x+l]) before analysis. Initially, there were 214 least disturbed sites for zooplankton. We
performed a linear regression of zooplankton MMI score versus the score for the first principal
component. Before calculating benchmarks, we performed a 1.5*IQR outlier analysis on the set
of least disturbed site MMIs to remove outliers. We excluded three sites based on this test (one
each in the CPL EHIGH, and WMTNS), leaving 211 least disturbed sites. Of the 211 least
disturbed sites, 9 sites (8 in WMTNS and 1 in PLAINS) were missing data required for the PCA
analysis, and so do not have principal component scores (mostly missing turbidity in CA). Thus,
there were a total of 202 sites used for the benchmark adjustment statistical analysis.
The best regression model had two different slopes and separate intercepts for each bio-region
(Table 7-10). The pooled model RMSE was 10.86. We used a pooled RMSE (based on all sites) to
provide an adequate sample size for estimating the distribution of MMI scores about the
intercept value for each bio-region. The regression models for the CPL, EHIGH and UMW bio-
regions had no relationship with disturbance and their slopes were set to zero. The slopes for
the PLAINS and WMTNS bio-regions were similar enough that a single value (-6.113) was used
for both. The intercepts were 74.16 in the CPL, 78.75 in the EHIGH, 74.10 in the UMW, 58.32 in
the PLAINS, and 74.39 in the WMTNS. Table 7-11 shows both the raw (unadjusted sample) 5th
and 25th percentiles and the regression model adjusted percentiles that we are using as the
MMI benchmarks. In three bio-regions (CPL, EHIGH, and UMW), the adjustment resulted in as
slight lowering (< 2 points) of the Good/Fair benchmark value. In the PLAINS and WMTNS bio-
regions, the Good/Fair benchmark values were increased (4.6 to 5.6 points). Adjustment
lowered the Fair/Poor benchmark values in the CPL, EHIGH, and UMW bio-regions by 2.7 to 6.7
points. The Fair/Poor benchmark value was increased by 14.5 points in the PLAINS bio-region,
and 3.9 points in the WMTNS bio-region.
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Least Disturbed Sites
6 m
(113) (97)
Depth Class
Figure 7.12,Zooplankton MMI scores versus site depth for least disturbed sites.
Upper panel shows MMI scores versus actual site depth. The reference line of 6 m separates shallower lakes
where the entire water column was sampled and deeper lakes where part of the water column was not
sampled. The lower panel compares distribution of MMI scores in shallow lakes (<6 m; n=113) versus deeper
lakes (> 6 m, n=97). Dots indicate the 5th and 95th percentiles.
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Table 7-10. LINEAR REGRESSION STATISTICS OF ZOOPLANKTON MMI SCORES VERSUS PCA-BASED DISTURBANCE
SCORE FOR EACH BIO-REGION.
Bio-Region
Slope
Intercept
RMSE (Pooled)
Coastal Plains (CPL)
0
64.94
10.01
Eastern Highlands (EHIGH)
0
76.50
10.01
Plains (PLAINS)
-6.143
54.55
10.01
Upper Midwest (UMW)
0
72.49
10.01
Western Mountains
(WMTNS)
-6.143
63.48
10.01
Table 7-11. ECOLOGICAL CONDITION BENCHMARKS FOR ZOOPLANKTON MMI SCORES (NLA 2012 ONLY) BASED ON
THE DISTRIBUTION OF LEAST DISTURBED SITES IN FIVE BIO-REGIONS.
Poor condition indicates a site is different from least disturbed condition. Fair condition indicates a site is
somewhat deviant from least disturbed condition. Good condition indicates a site is similar to least disturbed
condition. Values in bold (adjusted based on the regressions of MMI scores to PCA-based disturbance scores) are
used to assign condition.
Good/Fair Benchmark
Range of MMI
Bio-
(P25)
Fair/Poor Benchmark (P5)
scores in Least
Region
na
Adjusted
Unadjusted
Adjusted
Unadjusted
disturbed Sites
Coastal
22
SI J
59.4
48.4
49.7
38.80 to 94.47
Plains
(CPL)
Eastern
59
57.2
58.0
60.0
57.3
46.37 to 92.62
Highlands
(EHIGH)
Plains
37
42.4
37.8
33.2
17.4
4.42 to 78.57
(PLAINS)
Upper
31
73.3
73.7
56.0
58.0
53.37 to 92.01
Midwest
(UMW
Western
51
69.2
63.6
54.6
53.9
31.24 to 97.94
Mountains
(WMTNS)
0 Number of least disturbed sites remaining after excluding statistical outliers and sites with missing PCA -based
disturbance scores.
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7.7.2 NLA 2017
The process used to develop condition class benchmarks in the NLA 2012 was modified as
follows for the NLA 2017:
1. We excluded within-year revisits (see Section 2.1.3) and used the 2017 visit for sites that
were sampled in both 2012 and 2017.
2. We did not try to adjust the benchmarks for varying quality among regions by using the
"hindcasting" approach described by Herlihy et al. (2008).
Before calculating benchmarks for each of the five bio-regions, we removed outliers based on a
1.5*IQR outlier analysis of the MMI scores in least disturbed sites (Tukey 1977). We used the
25th percentile value to distinguish sites in "good" condition (similar to least disturbed) from
sites in "fair" condition (slightly deviant from least disturbed). We used the 5th percentile value
to distinguish sites in "fair" condition from sites in "poor" condition (different from least
disturbed). The revised benchmark values (Table 7-12) were used to assign condition classes for
the NLA 2017 sites, and to re-assign condition classes for the NLA 2012 sites (so that the change
in condition status could be estimated).
Table 7-12 Ecological condition benchmarks for NLA 2017 zooplankton MMI scores based on the distribution of
least disturbed sites in five aggregated ecoregions (bio-regions).
Poor condition indicates a site is different from least disturbed condition. Fair condition indicates a site is
somewhat different from least disturbed condition. Good condition indicates a site is similar to least disturbed
condition.
Number of
Least Disturbed
Zooplankton
Good-Fair
Fair-Poor
Bio-region
Sites"
Benchmark
Benchmark
Coastal Plains
23
59.42
53.77
Eastern
88
73.595
60.03
Highlands
Plains
61
36.72
28.17
Upper Midwest
61
63.68
52.03
Western Mountains
102
60.78
51.32
0 Based on a single visit per site from the NLA 2012 and the NLA 2017 and after excluding sites where less than 100
individuals were collected in either the coarse or fine net sample, anomalous samples, and statistical outliers.
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7.7.3 NLA 2022
Forthe NLA 2022, we used the same benchmarks for assigning condition class as those presented
in Table 7-12.
7.8 Discussion
We were able to develop regional MMIs for pelagic zooplankton assemblages that were
sufficiently responsive and repeatable to allow us to assess ecological condition for the NLA.
The zooplankton assemblage appears to be responsive principally to disturbance resulting from
increased nutrients and from increases in agricultural-related activity, which is consistent with
previous studies (e.g., Gannon and Stemberger 1978, Stemberger and Lazorchak 1994). We did
not observe a strong response of the zooplankton assemblage to shoreline habitat disturbance,
as has been noted by others (e.g., Stemberger and Lazorchak 1994).
Based on our evaluations, the zooplankton MMIs we developed do not appear to be affected by
lake origin (except possibly in the WMTNS), lake size, or by the use of a restricted tow length
that does not collect individuals which might be occupying waters deeper than 6 m. Presence of
these effects requires dealing with different types or sizes of lakes differently, either in terms of
developing separate MMIs for them, or in setting different benchmark values for them based
on a very small number of least disturbed lakes.
The regional zooplankton MMIs have the following limitations. Samples must be collected using
the same protocols and nets. Individuals were identified to the lowest practical taxon (with
species being the target level). However, total richness metrics did not perform well in terms of
responsiveness or repeatability, so coarser level identification may be possible in the future.
However, coarser-level identification will constrain the development of predictive models
based on taxa richness (O/E models) and would reduce the precision associated with biomass
estimates due to lumping of taxa to coarser levels. Many richness metrics didnot perform well
in the 2012 NLA, but stronger richness signals may be observed in future rounds of the NLA.
Many density- and biomass-based metrics did perform well, thus laboratory analyses will
require the determination of biomass, which increases costs.
In some bio-regions, our requirement for inclusion of at least one metric from each of the six
categories resulted in using metrics that were either not very responsive to disturbance or were
not very repeatable, and, in some bio-regions, including metrics that were most correlated.
Eliminating the poor-performing metrics from the suite of metrics did not appear to improve
the MMI performance, so we retained them for consistency across bio-regions. Moreover, in
those cases where we had a pair of highly correlated metrics, the mean correlation among all
pairs of component metrics was low, so we did not feel the correlation unduly influenced the
performance of the MMI (Van Sickle 2010). Future research might eliminate the requirement of
metric categories and just include the best performing metrics regardless of metric category to
determine if the resulting MMIs prove to be more responsive and repeatable than those
described in this document.
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We observed that the responses of some metrics were contradictory to what we expected with
increased disturbance (Table 7-1). However, little information is available, other than
generalization about taxa richness and assemblage composition, and possibly feeding ecology,
to support or refute the responses we observed in metrics related to density or biomass. We
are not aware of any studies that have conducted an evaluation of an exhaustive list of
candidate zooplankton metrics such as we developed for the NLA; it is possible that there has
not been the incentive to do so up to now. We hope that the success of the initial NLA
zooplankton MMIs will increase interest in the use of zooplankton metrics and indices in lake
bioassessment activities. This would lead to additional information related to responses of
zooplankton assemblages to various types of human disturbance.
We also worked with a limited set of autecological information for the zooplankton taxa that
were collected (essentially taxonomic and coarse-level feeding ecology). Additional information
is available for a limited number of taxa (e.g., Sprules and Holtby 1979, Barnett et al. 2007,
2013, Vogt et al. 2013; Hebert et al. (2016)), but it is uncertain if this information can be
assigned to related taxa. We did not have any information regarding the tolerance of
zooplankton taxa either to specific stressors or to a generalized disturbance variable. Tolerance
values have been developed for large numbers offish taxa as well as benthic invertebrate taxa
(Yuan 2004, Carlisle et al. 2007, Whittier et al. 2007, Meador et al. 2008, Whittier and Van Sickle
2010), and for rotifers in New Zealand (Duggan et al. 2001). Data are available from NLA 2007
that would allow tolerance values to be developed and applied to the NLA zooplankton MMI,
albeit at a coarser taxonomic level than species, and tolerance values derived from NLA 2012
would be available for future assessments.
Finally, it is well known that predation by fish and larger invertebrate predators can affect
zooplankton assemblages. Predation by planktivorous fish can result in smaller-sized taxa
becoming more abundant. The NLA does not collect any detailed information about fish
assemblages, so interpretations of response of metrics or the MMI to increased nutrients may
be confounded with an increase in the number of fish species (including planktivorous species)
that might accompany an increase in nutrients and a shift in the temperature regime from cold
water to warm water.
The primary modifications to the NLA zooplankton MMI indicator implemented for the NLA
2017 were focused on defining the reference distribution for ecological condition benchmark
calculations. Adding a minimum count criterion for excluding least disturbed sites before
calculating ecological condition benchmarks is consistent with what is done for the NLA benthic
macroinvertebrate MMI. We excluded more NLA 2012 sites with this screen than NLA 2017
sites. The observed decrease may have been due to clarifications made in the field NLA 2017
operations manual and during training to help reduce the occurrence of problematic samples.
For sites that were not least disturbed, we did not treat sites with low counts differently, unless
there was evidence that any zooplankton sample was compromised.
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We combined least disturbed sites from the NLA 2012 and the NLA 2017 to increase the sample
sizes to provide more robust estimates of the percentiles on which the condition class
benchmarks are based. This is consistent with what has been done for several other NLA
indicators that derive benchmarks based on least disturbed condition. Sample sizes were
substantially increased in four of the five bio-regions. The sample size for the Coastal Plains
(CPL; n=24) was only increased by two sites over what was available in the NLA 2012.
Finally, we have determined for other indicators and NARS assessments that adjusting the
percentiles used as benchmarks for ecological condition class assignments using the approach
described in Herlihy et al. (2008) does not yield benchmarks that are much different from the
unadjusted percentiles for nearly all aggregated ecoregions (or bio-regions). The adjustment
process requires additional time and effort and is more complicated to explain. Having
increased sample sizes of least disturbed sites from combining multiple surveys may be a factor
in the increased comparability of the unadjusted and adjusted percentiles.
Several aspects of the zooplankton MMI development process warrant further work:
1. Evaluating MMIs constructed using the best-performing metrics regardless of their
metric category.
2. Investigating metrics that perform well, but whose response to disturbance appears to
be contrary to our current expectations.
3. Developing better autecological information for zooplankton taxa, especially with
respect to tolerance to environmental stressors.
All of these aspects are still applicable after the NLA 2017 study and could lead to refinements
of the MMI process before the next round of the NLA is implemented in 2017.
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Gelinas, M., and B. Pinel-Alloul. 2008. Relating crustacean zooplankton community structure to
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Chapter 8: Human Health Water Quality Indicators
8.1 Enterococci indicator
The EPA developed and validated a molecular testing method employing quantitative
polymerase chain reaction (qPCR) as a rapid approach for the detection of enterococci in
recreational water (USEPA 2015). NLA used this method to estimate the presence and quantity
of these fecal indicator bacteria in the nation's lakes. The statistical benchmark value of 1280
calibrator cell equivalents (CCE)/100 mL from EPA's 2012 Recreational Water Quality Criteria
document (USEPA 2012)was then applied to the enterococci data to assess the recreational
condition of coastal waters.
8.1.1 Field collection
To collect enterococci samples, field crews took a water sample from the last littoral station or
the launch site in an area that was approximately 1 m deep at about 0.3 m (12 inches) below
the water. Following collection, crews placed the sample in a cooler and kept it on ice prior to
filtration of two 50 mL volumes. Samples were filtered and frozen on dry ice within 6 hours of
collection. The frozen filters were shipped to the laboratory on dry ice. A sterile phosphate
buffer solution (PBS) blank was also filtered at revisit sites durring one of the two visits.
8.1.2 Lab methods
The sample collections and the laboratory method followed EPA's Enterococcus qPCR Method
1609.1 (USEPA 2015);). Method 1609.1 describes a quantitative polymerase chain reaction
(qPCR) procedure for the detection of DNA from enterococci bacteria in ambient water
matrices based on the amplification and detection of a specific region of the large subunit
ribosomal RNA gene (IsrRNA, 23S rRNA) from these organisms. This method uses an arithmetic
formula (the comparative cycle benchmark (CT) method; Applied Biosystems, 1997) to calculate
the ratio of Enterococcus IsrRNA gene target sequence copies (TSC) recovered in total DNA
extracts from the water samples relative to those recovered from similarly prepared extracts of
calibrator samples containing a consistent, pre-determined quantity of Enterococcus cells.
Mean estimates of the absolute quantities of TSC recovered from the calibrator sample extracts
were then used to determine the quantities of TSC in the water samples and then converted to
CCE values as described in the section below. To normalize results for potential differences in
DNA recovery, monitor signal inhibition or fluorescence quenching of the PCR analysis caused
by a sample matrix component, or detect possible technical error, CT measurements of sample
processing control (SPC) and internal amplification control (IAC) target sequences were
performed as described in Method 1609.1.
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8.1.3 Analysis and application of benchmarks
8.1.3.1 Calibration
Estimates of absolute TSC recoveries from the calibrator samples were determined from
standard curves using EPA-developed plasmid DNA standards of known TSC concentrations as
described in Method 1609.1. Estimates of TSC recovered from the test samples were
determined by the comparative cycle threshold (CT) method, as also described in Method
1609.1. Before applying the EPA benchmark to the qPCR data, it was necessary to convert the
TSC estimates to CCE values.
The standardized approach developed for this conversion is to assume 15 TSC/CCE (USEPA
2015). This approach allows the CCE values to be directly compared to the EPA RWQC values
(Haugland et al., 2014). A slightly modified approach was employed in the earlier NRSA 2008-09
study to obtain the same conversions of TSC to standardized CCE units.
8.1.3.2 Benchmarks
For the data analysis of the enterococci measurements determined by Method 1609.1, analysts
used a benchmark as defined and outlined in EPA's recommended recreational criteria
document for protecting human health in ambient waters designated for swimming (USEPA
2012). Enterococci CCE/100 mL values were compared to the EPA benchmark of 1280 CCE/100
mL.
Within-year sampling variability was assessed by comparing NLA 2022 visit 1 and 2 condition
categories and is presented in Table 8-1. For conditions categories of "at or below benchmark",
"above benhcmark" and "not assessed", results showed agreement in 84 (87.5%) of the 96
revisit sites sampled in 2022.
Table 8-1 Enterococci condition contingency table; N = 96.
Enterococci Condition
Visit 1
At or Below Benchmark
Above Benchmark
Not Assessed
At or Below Benchmark
82
5
1
Visit 2
Above Benchmark
6
2
Not Assessed
8.2 Cyanobacteria toxins (Cyanotoxins)
Cyanobacteria are one-celled photosynthetic organisms that normally occur at low levels.
Under eutrophic conditions, cyanobacteria can multiply rapidly. Not all cyanobacterial blooms
are toxic, but some may release toxins, such as microcystins and cylindrospermopsin. For the
NLA, both microcystins and cylindrospermopsin were analyzed.
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Recreational exposure is typically a result of inhalation, skin contact, or accidental ingestion.
When people are exposed to cyanotoxins, adverse health effects may range from a mild skin
rash to serious illness or in rare circumstances, death. Acute illnesses caused by short-term
exposure to cyanobacteria and cyanotoxins during recreational activities include hay fever-like
symptoms, skin rashes, respiratory and gastrointestinal distress.
Microcystins refers to an entire group of toxins (all of the different congeners, rather than just
one congener). Cyanobacteria can produce one or many different congeners at any one time,
including Microcystin - LR (used in the kit's calibration standards), Microcystin - LA, and
Microcystin - RR. The different letters on the end signify the chemical structure (each one is
slightly different) which makes each congener different.
8.2.1 Field methods
Samples for cyanotoxin analyses were collected using a 0-2 m vertically integrated water
column sampler at the open-water site. Water from the photic zone was emptied into a 4L
cubitainer and then transferred to a 500 mL bottle. The bottle was kept on ice and then stored
frozen until analysis. Both microcystins and cylindrospermopsin were analyzed from the 500 mL
bottle.
8.2.2 Analysis and application of benchmarks
Microcystins were measured using an enzyme-linked immunosorbent assay (ELISA) procedure
with an Abraxis Microcystins-ADDA Test Kit. For freshwater samples, the procedure's reporting
range is 0.15 n g/L to 5.0 n g/L and the minimum detection level (MDL) is 0.10 |-ig/L.
Microcystins concentrations were evaluated against the EPA recommended criterion and
swimming advisory level of 8 |ag/L (USEPA 2019).
The cylindrospermopsin sample was measured using an enzyme-linked immunosorbent assay
(ELISA) procedure with an Abraxis Cylindrospermopsin Test Kit. For freshwater samples, the
procedure'sreporting range is 0.02 n g/L to 2.0 n g/L and the MDL is 0.04 |-ig/L.
Cylindrospermopsin concentrations were evaluated against the EPA recommended criterion
and swimming advisory level of 15 |ag/L (USEPA 2019).
The NLA also reports on the percentage of lakes with detections of cyanotoxins and changes in
detection over time. Detection is defined as a value greater than the MDL. When the MDL
changed between surveys, the greatest MDL for all surveys is used to determine detect/not
detected.
Within-year sampling variability for microcystin condition was assessed by comparing NLA
2012, 2017 and 2022 visit 1 and 2 condition categories and is presented in Table 8-3Table 8-1.
For microcystin detection, results showed agreement in 221 (75%) of the 293 revisit sites
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sampled over three survey years. For microcystin risk condition, 289 (98%) of the 293 revisit
sites over three survey years were in agreement.
Table 8-2. Microcystin detection (a) and risk condition (b) contingency tables; N = 293.
a)
Microcystin Detection
Visit 1
Detected
Not-detected
Not Assessed
Visit 2
Detected
72
39
Not detected
33
148
Not Assessed
1
b)
Microcystin Risk Condition
Visit 1
At or Below
Benchmark
Above
Benchmark
Not
Assessed
At or Below
Benchmark
287
2
Visit 2
Above Benchmark
2
1
Not Assessed
1
Within-year sampling variability for cylindrospermopsin was assessed by comparing NLA 2017
and 2022 visit 1 and 2 condition categories and is presented in Table 8-3Table 8-1. For
detection, results showed agreement in 177 of the 193 revisit sites. For risk condition, 100% of
the risk categores were the same.
Table 8-3. Cylindrospermopsin detection (a) and risk condition (b) contingency tables; N = 193.
a)
Cylindrospermopsin Detection
Visit 1
Detected
Not-detected
Not Assessed
Visit 2
Detected
12
11
Not detected
4
165
Not Assessed
1
b)
Cylindrospermopsin Risk Condition
Visit 1
At or Below
Benchmark
Above
Benchmark
Not
Assessed
Visit 2
At or Below
Benchmark
192
Above Benchmark
Not Assessed
1
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8.3 Literature cited
Abraxis, Cylindrospermopsin ELISA Microtiter Plate Enzyme-Linked Immunosorbent Assay for
the Determination of Cylindrospermopsin in Water," Product 520011, UG 21-059 (REV
01), Undated. Retrieved December 2022 from
https://www.goldstandarddiagnostics.us/media/15642/ug-21-059-rev-01-abraxis-
cylindrospermopsin-elisa 522011.pdf.
Abraxis, "Cylindrospermopsin ELISA Plate 522011," Flowchart. 03FEB2022. Retrieved December
2022 from https://www.goldstandarddiagnostics.us/media/15789/fc-22-043-rev-01-
abraxis-cylindrospermopsin 522011.pdf.
Abraxis, "Microcystins-ADDA ELISA Microtiter Plate Enzyme-Linked Immunosorbent Assay for
the Congener-Independent* Determination of Microcystins and Nodularins in Water
Samples," Product 520011, UG 21-052 (REV 01), Undated. Retrieved December 2022
from https://www.goldstandarddiagnostics.us/media/15635/ug-21-052-rev-01-abraxis-
microcystins-adda-elisa 520011.pdf.
Abraxis, "Microcystin-ADDA ELISA Plate 520011," Flowchart. 03FEB2022. Retrieved December
2022 from https://www.goldstandarddiagnostics.us/media/15783/fc-22-037-rev-01-
abraxis-microcystin-adda 520011.pdf.
Applied Biosystems (1997) User Bulletin #2. ABI PRISM 7700 Sequence Detection System.
Applied Biosystems Corporation, Foster City, CA.
Haugland, R.A., S.D. Siefring, M. Varma, A.P. Dufour, K.P. Brenner, T.J. Wade, E. Sams, S.
Cochran, S. Braun, and M. Sivaganesan. 2015. Standardization of enterococci density
estimates by EPA qPCR methods and comparison of beach action value exceedances in
river waters with culture methods. J. Microbiol. Methods. 105, 59-66.
James, R., et al., "Environmental Technology Verification Report: Abraxis Microcystin Test Kits:
ADDA ELISA Test Kit; DM ELISA Test Kit; Strip Test Kit," in Environmental Technology
Verification System Center 2010. Retrieved March 2013 from
http://nepis.epa.gov/Adobe/PDF/P100EL6B.pdf
USEPA. 2012. Recreational Water Quality Criteria. EPA 820-F-12-058. Washington, D.C.
USEPA. 2015. Method 1609.1: Enterococci in water by TaqMan® quantitative polymerase chain
reaction (qPCR) assay with internal amplification control (IAC) assay. EPA-820-R-15-009.
US Environmental Protection Agency, Office of Water, Washington, D.C. Available on-
line at https://www.epa.gov/sites/default/files/2015-08/documents/method 1609-1-
enterococcus-iac 2015 3.pdf
USEPA. 2019. Recommended Human Health Recreational Ambient Water Quality Criteria or
Swimming Advisories for Microcystins and Cylindrospermopsin. EPA 822-R-19-001. U.S.
Environmental Protection Agency, Office of Water, Washington, DC.
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Chapter 9: Human Health Fish Tissue Indicators
Fish are time-integrating indicators of persistent pollutants, and the bioaccumulation of contaminants in
fish tissue has important human health implications. Contaminants in fish pose various health risks to
human consumers (e.g., cancer risks, and noncancer risks such as reproductive effects or impacts to
neurological development). The NLA 2022 human health fish tissue indicator consists of the collection of
whole fish samples for homogenized fillet analyses. These samples provide information on the national
distribution of selected persistent, bioaccumulative, and toxic (PBT) chemical residues (specifically,
mercury, polychlorinated biphenyls, or PCBs, and per- and polyfluoroalkyl substances, or PFAS) in fish
species that people might catch and eat. Results of analyses of mercury, PCB, and PFAS fillet tissue
concentrations are presented for this indicator.
9.1 Field fish collection
The human health fish tissue indicator field and analysis procedures described below were based on the
EPA's National Study of Chemical Residues in Lake Fish Tissue (USEPA 2009) and the EPA's Guidance for
Assessing Chemical Contaminant Data for Use in Fish Advisories, Volumes 1-2 (third edition) (USEPA
2000a).
The NLA crews attempted to collect whole fish samples for the fillet tissue indicator from a subsample of
approximately two-thirds of the lakes included in the survey design. In total, 413 whole fish composite
samples were collected from the 636 designated lakes in the lower 48 states. Each lake had a surface
area >1 hectare and contained at least 1,000 square meters of open, unvegetated water and a
permanent population of predator fish species. The fish samples collected for fillet tissue analysis
consisted of a composite of predator fish specimens1 from each site. Additional criteria for each fish
composite sample included fish that were:
• All of the same fish species that are commonly caught and consumed by humans,
• Harvestable size per legal requirements or of consumable size if there were no harvest limits,
• At least 190 mm in length and of similar size so that the smallest individual in the composite was
no less than 75% of the total length of the largest individual in the composite, and
• Sufficiently abundant within the lake.
Crews were provided with a recommended list of primary and secondary fish species (Table 9-1), but
they could choose an appropriate substitute (based on the criteria listed above) if none of the
recommended fish species were available. Fish collection data were screened to exclude individual fish
specimens with lengths less than 190 mm or composite samples where field crews collected non-target
or unacceptable substitute species.
To prepare fillet composite samples for chemical analysis, fish composite samples from each site were
scaled and filleted in the laboratory. In filleting individual fish, muscle tissue was removed from both
sides of each fish leaving the skin on and the belly flap attached to the fillet. Fillets from the individual
specimens that comprised a composite sample were homogenized together before being analyzed for
contaminants.
1 Use of composite sampling for screening studies is a cost-effective way to estimate average contaminant
concentrations while also ensuring that there is sufficient fish tissue to analyze for all contaminants of concern and
to archive surplus tissue, when possible.
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Table 9-1. 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
9.2 Mercury analysis and fish tissue screening levels to protect human health
All fish tissue samples were analyzed for total mercury. The samples were prepared using EPA Method
1631B, Appendix A (USEPA 2001a) and analyzed using EPA Method 1631E (USEPA 2002), which utilizes
approximately 1 g of fillet tissue for analysis. In screening-level studies of fish contamination, the EPA
guidance recommends monitoring for total mercury rather than methylmercury (an organic form of
mercury) because most mercury in adult fish is in the toxic form of methylmercury which will be
captured during an analysis for total mercury. Applying the assumption that all mercury is present in fish
tissue as methylmercury is a conservative approach protective of human health.
The fish tissue criterion used to interpret mercury concentrations in fillet tissue for human health
protection is 0.3 milligrams (mg) of methylmercury per kilogram (kg) of tissue (wet weight), or 300 parts
per billion (ppb), which is EPA's fish tissue-based CWA Section 304(a) water quality criterion
recommendation for methylmercury (EPA 2001b).2 For more information on the screening levels for
human health protection, see Section 9.5.
2 Because the EPA relies on the recommended CWA Section 304(a) national water quality criterion for
methylmercury to interpret the mercury results, the EPA is only reporting mercury results for general population
and is not including an additional analysis and interpretation of mercury results for high-frequency fish consumers
or reduced-frequency fish consumers.
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Application of this criterion to the mercury fillet tissue composite data from this study identifies the
proportion of lakes in the sampled population containing fish with mercury fillet tissue concentrations
that are above the criterion. Mercury concentration data from analysis of homogenized fish fillet
composite samples are available to download from the NLA Fish Tissue Study webpage. Summary
statistics, including the number of detections, are reported in Table 9-2, and the proportion of lakes with
sample exceedances above the mercury criterion is reported in Table 9-3.
9.3 PCB analysis and fish tissue screening levels to protect human health
All fish tissue samples were analyzed for PCBs. EPA Method 1668C (USEPA 2010) was used to analyze
approximately 10 g of homogenized fillet tissue from each fish composite sample to provide results for
the full suite of 209 PCB congeners. The total PCB concentration for each sample was determined by
summing the results for any of the 209 congeners that were detected, using zero for any congeners that
were not detected in the sample.
In the main report, National Lakes Assessment: The Fourth Collaborative Survey of Lakes in the United
States, the EPA included total PCB results for general fish consumers (those who may eat one 8-ounce
meal of locally caught fish per week), for high-frequency fish consumers (those who may eat four or five
8-ounce meals of locally caught fish per week), and for reduced-frequency fish consumers (those who
may eat one 8-ounce meal of locally caught fish per month). The EPA used fish tissue screening levels,
expressed as wet-weight concentrations of total PCBs, to protect human health by characterizing cancer
human health risks for these three levels of fish consumers. For more information on the fish tissue
screening levels for human health protection, see Section 9.5.
Application of these screening levels to the PCB fillet tissue data identifies the proportion of lakes in the
sampled population containing fish with total PCB fillet concentrations that are above each total PCB
fish tissue screening level. PCB concentration data from the analysis of homogenized fish fillet
composite samples are available to download from the NLA Fish Tissue Study webpage. Summary
statistics, including the number of detections, are reported in Table 9-2, and the proportion of lakes with
sample exceedances above each screening level for three levels offish consumers is reported in Table 9-
3.
9.4 PFAS analysis and results
All fish tissue samples were analyzed for 40 per- and polyfluoroalkyl substances (PFAS), using EPA
Method 1633 (USEPA 2024a). This method, which utilizes approximately 2 g of fillet tissue for analysis,
uses high performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) and
applies isotope dilution to determine the concentration of each of the 40 PFAS.
In the main report, National Lakes Assessment: The Fourth Collaborative Survey of Lakes in the United
States, EPA reports results on frequencies of detection of the most commonly detected PFAS (i.e., those
PFAS that were detected in at least 20 percent of the fillet tissue samples). In addition, EPA reports the
estimates of the number of lakes in the sampled population containing fish with detectable levels of
PFAS. PFAS concentration data from fish fillet tissue composite samples are available to download from
the NLA Fish Tissue Study webpage. Summary statistics for PFAS, including the number of detections for
each of the 40 tested PFAS are provided in Table 9-2.
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Table 9-2. NLA 2022 fish tissue fillet composite sample summary data
Chemical
Number of
Detection
MDL (ppb)a
Measured
Weighted
Measured
Detections
Frequency
Minimum
Median
Maximum
(%)
Concentration
(ppb)b
Concentration
(ppb)
Concentration
(ppb)
Mercury
413
100
0.8
4.5
308.956
1660
Total PCBs
413
100
0.000134-
0.000797
0.013
0.958
131.482
Perfluoroalky
carboxylic acids
PFBA
3
0.73
0.372
0.472
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Chemical
Number of
Detection
MDL (ppb)a
Measured
Weighted
Measured
Detections
Frequency
Minimum
Median
Maximum
(%)
Concentration
(ppb)b
Concentration
(ppb)
Concentration
(ppb)
ADONA
0
0
0.384
0
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Each screening level is expressed as a fish fillet tissue contaminant level (ng/g or ppb) that, if exceeded,
may adversely impact human health among people who eat a specified amount offish. See Table 9-3 for
the chemical- and fish consumption-specific screening levels and the estimated portion of lakes that
contained fish with fillet tissue that exceeded screening levels corresponding to each contaminant and
fish consumption rate.
Because PCBs can have both cancer and noncancer health effects, the EPA calculated fish tissue
screening levels for each type of health effect and used the lower of the two screening levels. For each
rate of fish consumption (general fish consumers, high-frequency fish consumers and reduced-
frequency fish consumers), the EPA developed two fish tissue screening levels for PCB human health
impacts for the purpose of directly comparing to fish fillet tissue results - one based on noncancer
effects, and one based on cancer effects. The screening levels represent the concentration of total PCBs
in fish tissue that should not be exceeded based on three levels of fish consumption rates ranging from
0.142 kg of fish/day (for high-frequency fish consumers who may consume four or five 8-ounce serving
of locally caught fish per week) to 0.0324 kg of fish/day (for general fish consumers who may consume
one 8- ounce serving of locally caught fish per week)to 0.00745 kg of fish/day (for reduced-frequency
fish consumers those who may eat one 8-ounce serving of freshwater fish per month). The PCB
screening levels were also based on a human adult body weight default value of 80 kg3 and a RfD of
0.00002 mg/kg day or a cancer slope factor of 2 (mg/kg/d)1 (USEPA 1994) and a cancer risk level of 10"5.
For the screening level for general fish consumers, EPA used a fish consumption rate of 32 grams per day
(or one 8-ounce meal of locally caught river fish per week), consistent with the U.S. Department of
Agriculture and Department of Health and Human Services' Dietary Guidelines for Americans, 2020-
2025 (USDA and HHS 2020). For the screening level for high-frequency fish consumers (such as
subsistence or recreational fishers or individuals from underserved populations), EPA used a fish
consumption rate of 142 grams per day (or four to five 8-ounce meals of locally caught river fish per
week) which is described in the EPA 2000 Human Health Methodology (USEPA 2000b). Because the total
PCBs screening levels associated with cancer effects were lower than the screening levels associated
with noncancer effects, the EPA applied only the screening levels associated with cancer effects. This
conservative approach is also likely to be protective against noncancer effects, which may occur at
higher levels of total PCB contamination.
Table 9-3. NLA 2022 fish fillet tissue sampled population exceedances for mercury and total polychlorinated biphenyls (PCBs)
Chemical
Percent
Detection, %
Percent of Lakes in the Sampled Population with Fish that Exceeded the Mercury Criterion Level, %
(ppb. Note: 1 ppb = 1 ng/g)
Mercury
100%
51%
(300 ppb)
Chemical
Percent
Detection, %
Percent of Lakes in the Sampled Population with Fish that Exceeded the Total PCBs Screening Levels
for Different Levels of Fish Consumers, %
(Calculated Screening Level, ppb. Note: 1 ppb = 1 ng/g)
High-Frequency Consumer
General Consumer
Reduced-Frequency Consumer
Four to five
8-oz meals/week
One 8-oz meal/week
One 8-oz meal/month
Total PCBs
100%
23%
(2.8 ppb)
6%
(12 ppb)
2%
(54 ppb)
3 The EPA's toxicity assessment for PCBs summarizes health effects to the general population of adults over a
lifetime of exposure, so a national default estimated body weight for adults was used to derive screening levels for
PCBs (see Exposure Factors Handbook, Chapter 8, Table 8-1).
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9.6 Literature cited
USEPA. 1994. Integrated Risk Information System (IRIS) Chemical Assessment Summary for
Polychlorinated biphenyls (PCBs); CASRN 1336-36-3. U.S. Environmental Protection Agency,
National Center for Environmental Assessment, Washington, DC
USEPA. 2000a. Guidance for Assessing Chemical Contaminant Data for Use in Fish Advisories, Volumes 1-
2 (Third Edition). EPA 823-B-00-007. U.S. Environmental Protection Agency, Office of Water,
Washington, DC.
USEPA. 2000b. Methodology for Deriving Ambient Water Quality Criteria for the Protection of Human
Health (2000). EPA 822-B-00-004. U.S. Environmental Protection Agency, Office of Water,
Washington, DC.
USEPA. 2001a. Appendix to Method 1631, Total Mercury in Tissue, Sludge, Sediment, and Soil by Acid
Digestion and BrCI Oxidation. EPA-821-R-01-013. January 2001. U.S. Environmental Protection
Agency, Office of Water, Washington, DC.
USEPA. 2001b. Water Quality Criterion for the Protection of Human Health: Methylmercury. EPA-823- R-
01-001. U.S. Environmental Protection Agency, Office of Water, Washington, DC.
USEPA. 2002. Method 1631, Revision E: Mercury in Water by Oxidation, Purge and Trap, and Cold Vapor
Atomic Fluorescence Spectrometry. EPA-821-R-02-019. August 2002. U.S. Environmental
Protection Agency, Office of Water, Washington, DC.
USEPA. 2009. The National Study of Chemical Residues in Lake Fish Tissue. EPA-823-R-09-006. U.S.
Environmental Protection Agency, Office of Water, Washington, DC.
USEPA. 2010. Method 1668C, Chlorinated Biphenyl Congeners in Water, Soil, Sediment, Biosolids, and
Tissue by HRGC/HRMS, April 2010. U.S. Environmental Protection Agency, Office of Water,
Washington, DC.
U.S. Department of Agriculture (USDA) and U.S. Department of Health and Human Services (HHS).
Dietary Guidelines for Americans, 2020-2025. 9th Edition. December 2020. Available at
DietaryGuidelines.gov
USEPA 2024. Method 1633, Analysis of Per- and Polyfluoroalkyl Substances (PFAS) in Aqueous, Solid,
Biosolids, and Tissue Samples by LC-MS/MS. EPA-821-R-02-019. August 2002. U.S.
Environmental Protection Agency, Office of Water, Washington, DC.
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Chapter 10: From Analysis to Results
10.1 Background information
In the NLA 2022 public report, lake condition estimates based on chemical, physical and
biological information are expressed as percentage of lakes or number of lakes; therefore, site
weights from the probability design must be used to generate population estimates along with
the data from the probability sites sampled (981). Extent estimates for biological indicators and
other measures are used to calculate relative and attributable risk.
10.2 Population estimates
The survey design for the NLA, discussed in Chapter 2 of this document, produces a spatially
balanced sample using the NHDPIus HR for 1-5 ha lakes as the sampling frame. Each lake has a
known probability of being sampled (Stevens and Olsen 1999, Stevens and Olsen 2000, Stevens
and Olsen 2004). A sample weight is assigned to each individual site as the inverse of the
probability of that lake being sampled. Sample weights can be adjusted for different survey
populations (e.g., sampled population or target population; see Chapter 2 and Appendix B) and
are expressed as number of lakes. In 2017, EPA determined it was appropriate to adjust the site
weights used to calculate the population estimates to represent the percentage of lakes
relative to the target population and continued to present the results this way in 2022. Results
presented in NLA 2007 and 2012 were relative to the sampled population.
The probability of a site being sampled was related to lake size class and was stratified by state.
Site weights for the survey were adjusted to account for additional lakes (i.e., oversample lakes)
that were evaluated when the primary lakes were not sampled (e.g., due to denial of access,
being non-target). These weights are explicitly used in the calculation of lake condition and
extent estimates, so results can be expressed as estimates of lakes (i.e., numbers of lakes or
percentage of the entire resource) in a particular condition class for the entire contiguous U.S.
For examples of how this has been done for other National Aquatic Resource Survey (NARS)
assessments, see USEPA (2006), Olsen and Peck (2008), and USEPA (2009). It is important to
note that the NLA was not designed to report on individual lakes or states, but to report at
national and regional scales. The NLA 2022 national results are the focus in the public report.
Regional results are also presented for some indicators. All regional scale and subpopulation
results are presented in the interactive dashboard.
10.2.1 Subpopulations
10.2.1.1 Ecoregions
The EPA has defined ecoregions at various scale, ranging from the coarse ecoregions at the
continental scale (Level I) to finer ecoregions that divide the land into smaller units (Level II or
IV). The nine ecoregions used in NLA are aggregations of the Level III ecoregions delineated by
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EPA for the continental U.S. These nine ecoregions include the Northern Appalachians (NAP),
Southern Appalachians (SAP), Coastal Plains (CPL), Upper Midwest (UMW), Temperate Plains
(TPL), Southern Plains (SPL), Northern Plains (NPL), Western Mountains (WMT), and Xeric (XER).
Additional information on the NLA ecoregions is available on the NARS website.
10.2.1.2 Lake origin: natural vs. human-made
The NLA condition estimates can also be explored and analyzed by lake origin. Unfortunately,
there is not a clear dichotomy between natural and human-made lakes. Many naturally existing
lakes are altered hydrologically to widely varying degrees by flow control structures, lake level
augmentation, and other human activities. For NLA analyses, we defined human-made lakes as
only those that are totally artificial, either impounded streams/rivers (reservoirs) or excavated
basins, an adaption of the definition developed by Whittier et al. (2002) during the EMAP lake
surveys. Excavated lakes are formed by flooding of quarries, borrow pits or any other type of
human dug hole and usually lack flowing outlets. Impoundments were originally lotic
waterbodies now turned into lentic waterbodies intentionally by humans. In our definition for
NLA purposes, human-made lakes are those where no lake existed prior to European
settlement. These include millponds, created residential, agricultural, or recreational ponds and
lakes, as well as reservoirs created for flood control, water supply, or hydroelectric production.
Every other type of lake is considered natural, even if the flow or shape is highly altered by
humans.
It was not always easy to assign lake origin to NLA sample lakes. The following information was
used after sampling to determine the classification for each lake:
• Lake name (reservoir in name);
• Google Earth views (or ArcGIS Explorer Desktop);
• Online topographic maps (ArcGIS Explorer Desktop or DeLorme Topo USA);
• Field collected data (i.e., assessment form including determination of
Seepage/Drainage/Reservoir and dams; verification form with general comments;
maximum lake depth);
• Initial site evaluation/reconnaissance determination;
• GNIS waterbody type;
• Internet lake history searches; and
• Ecoregion location.
The process used to determine lake origin in NLA has evolved based in part on lessons learned
and in part due to advances in technology (e.g., availability of online images and maps and free
apps such as Google Earth and ArcGIS, Desktop Explorer). As a first step, we look for agreement
of the initial reconnaissance with the field crew classification, followed by a quick map or
Google Earth review. When there are discrepancies in this information, a more in depth analysis
was conducted. No one source of information on lake origin by itself, was definitive. Sources
sometimes give conflicting answers; therefore, we used a weight of evidence approach to make
the classification in difficult cases. General guidelines included the following.
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1. Ecoregion location review. The Southern Appalachians have almost no natural lakes, any
lake there classified as natural should be checked. Natural lakes are common in
glaciated ecoregions and less common elsewhere. In NLA in the past, the ratio of
human-made to natural lakes is about 1:1.
2. Google Earth review. Google Earth views are good to get the lay of the land and look for
obvious dams and human activities/roads around the lake. A lake with no roads or
human activity around them are unlikely to be human-made. We examined digital
topographic maps for dams, or other evidence of impoundment such as a substantial
elevation drop from lake surface to the outlet stream, a straight shoreline, or a road
crossing at the outlet.
3. Comparison of the mapped elevation change at the outlet to the maximum lake depth.
If maximum lake depth is greater than any possible dam/elevation change, it's not a
human-made impoundment by our definition.
4. Historical information search. Most named lakes have a surprising amount of
information about them on the internet (note this doesn't work well for very small
lakes, or lakes with no name).
Some common types of lakes were especially problematic when assigning lake origin.
Oxbow/riverine flood plain lakes. Classic oxbow lakes are inherently natural. However, many
old oxbow lakes or lakes in riverine floodplain are highly altered by human activities (e.g.,
road/railroad berms, bridges, dikes) and look very artificial. Unless we could tell that these lakes
were actually created (dug out) by humans, we classified them all as natural.
Wetland complex lakes. A number of lakes are part of wetland complexes. Many of them are in
areas heavily managed by humans (e.g., state and federal wildlife/wetland management areas).
These lakes are often very shallow and augmented to hold more water, and the flows are highly
regulated for purposes of wetland management. Whether these lakes met the NLA definition of
a lake (< 1 m deep and 10,000 m2 of open water) in the past or not is almost impossible to
determine. It's likely that in the past, some of these were what we now call wetlands and were
not NLA target lakes. We have, however, classified all of these types of lakes as natural in that
there was very likely some type of a wetland/waterbody there in the past, pre-human
development.
Augmented natural lakes. Many natural lakes are flow altered by human activity either by
outlet flow control or raising the height of the lake with some kind of dam. The dams on these
lakes are often very apparent when looking through the various sources of lake information,
but we consider these to be natural lakes if a lake basin existed their pre-human settlement. It
can be difficult to determine if a lake basin existed in the past to separate them from what we
define as human-made impoundments. Dam height (or elevation contours) versus lake depth
was one approach to differentiate the two as well as doing internet history searches.
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Irrigation/water district management. Water is often stored and moved around for irrigation or
drinking water, especially in the Xeric West. A number of lakes are a part of water management
districts where water is pumped into and out of them depending on water needs. If the lake
existed in the past, even though the flow now is extremely altered by humans, we called them
natural.
Quarry lakes, borrow pits, and reclaimed strip mine lakes. There are a large number of quarry
or borrow pit lakes and ponds that are created by humans when they dug holes and then the
holes filled with water. Since they are small and often unnamed it can be very hard to
distinguish these from small unnamed natural ponds. Looking at the general landscape, lake
shape, depths and crew notes are the only way to make an educated guess. For lakes within
reclaimed strip mines, topographic maps may provide more information than imagery). If a
major road (especially an interstate highway) is adjacent, road fill was often dug out from
adjacent areas creating the borrow pit. Larger borrow pits and big quarries are sometimes
turned into parks and have historical information.
10.3 Lake extent estimates
The condition of each NLA probability site (i.e., good, fair, poor; above or below benchmark;
detected or not-detected) is determined by the appropriate indicator values and benchmarks
established for that indicator and ecoregion. Next, the site weights from the probability design
are summed across all sites in each condition class to estimate the percentage of lakes
nationally or in other sub populations (e.g., ecoregions, natural vs. manmade lakes, etc) in each
condition class for the target population. The survey design allows calculation of confidence
intervals around these condition estimates and allows for estimates of the whole resource not
just those lakes sampled. Note that only Visit 1 (i.e., the index visit) data and only probability
sites are used in the calculation of extent. Hand-selected sites have a weight of zero. Using this
method, the lakes in a particular condition class is estimated and reported in percentage of
lakes or number of lakes.
10.4 Stressor extent, relative risk, and attributable risk
A major goal of the National Aquatic Resource Surveys is to assess the relative importance of
stressors that impact aquatic biota on a national basis. The EPA assesses the influence of
stressors in three ways: stressor extent, relative risk, and population attributable risk. In NLA,
each targeted and sampled lake was classified as being in either Good, Fair, or Poor condition,
separately for each stressor variable and for each biological response variable. From this data,
we estimated the stressor extent (prevalence) of lakes in Poor condition for a specified stressor
variable. We also estimated the relative risk of each stressor for a biological response. Relative
risk is the ratio of the probability of a poor biological condition when the stressor is poor to the
probability of a poor biological condition when the stressor is not poor (Van Sickle et al. (2006)).
Finally, we estimated the population attributable risk (AR) of each stressor for a biological
response. AR combines RR and stressor extent into a single measure of the overall impact of a
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stressor on a biological response, over the entire population of lakes (Van Sickle and Paulsen
(2008)).
10.4.1 Stressor extent
For each particular stressor, the stressor extent (SE) may be reported as the number of lakes,
the proportion of lakes, or the percent of lakes in Good, Fair, Poor, or Not Assessed condition. If
the SE is reported as the proportion of lakes, then it can be interpreted as the probability that a
lake chosen at random from the population will be in Poor condition for the stressor.
Stressor extent in Poor condition is estimated as
(1) SEP, the sum of the sampling weights for sites that are assessed in Poor condition
r
SEp — Wpj
i=l
/
(2) SEPp, as the ratio of the sums of the sampling weights for the probability selected sites
that are assessed in Poor condition divided by the sum of the sampling weights of all the
selected sites regardless of condition, i.e.,
SEP -
p~ iu*t
, or
(3) SERp, the percent of stressor extent in Poor condition (i.e., stressor relative extent)
SER„ == 100 * SEP„ = 100 *
ynp
^i=1
W.
PI
p p Z?=1W,
where wpi is the weight for the /'th selected site in the Poor condition category, Wj is the weight
for the /'th selected site regardless of condition category, np is the number of selected sites that
are in Poor condition, and n is the total number of sites regardless of their condition category. A
stressor condition category may use other terminology to identify if a site is in poor condition
but generically, we use the term Poor. Note that the extent for a response variable is defined
similarly.
10.4.2 Relative risk and attributable risk
To estimate relative risk and attributable risk, we restrict the sites to those that both the
stressor and response variable assessed as Good, Fair, or Poor (or their equivalents). That is, if a
site is Not Assessed for either the stressor or response variable, it is dropped. Next, for these
sites the condition classes are combined to be either Poor or Not Poor for the stressor and
response variables. For example, Not Poor combines the Good and Fair condition classes. Thus,
each sampled lake was designated as being in either Poor (P) or Not Poor (NP) condition for
each stressor and response variable separately.
To estimate the relative risk and attributable risk for one stressor (S) and one response (B)
variable, we compiled a 2x2 table (Table 10-1), based on data from all lakes that were included
in the probability sample and that had both the stressor and response variable measured. A
separate table must be compiled for each pair of stressor and response variables.
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Table 10-1. Extent estimates for response and stressor categories
# Response(B)
# Stressor (S)
# Not Poor (NP)
# Poor (P)
# Not Poor (NP)
# a = wnni
# b = ^wnpi
# Poor(P)
# c = J^wvni
# d=^wppi
Table entries (a, b, c, d) are the sums of the sampling weights of all sampled lakes that were
found to have each combination of Poor or Not Poor condition for stressor and response. For
example, d = wppi where npp is the number of sites with both the stressor and response
in poor condition and wppi is the weight for the /'th site. Note that the estimates in Table 10-1
may differ from the stressor extent estimates since both the stressor and response variables
must be measured at each site.
10.4.3 Relative risk
Relative risk (RR) is the ratio of the probability of a Poor biological condition when the stressor
is Poor to the probability of a Poor biological condition when the stressor is Not Poor. That is,
Pr(B = P\S = P)
RR ~ Pr(B = P\S = NP)
Using the simplified notation in Table 10-1, relative risk (RR) is estimated as:
d/(b + d)
est c/(a + c)
A RR = 1.0 indicates there is no association between the stressor and response. That is, a Poor
response condition in a lake is equally likely to occur whether or not the stressor condition is
Poor. A RR > 1.0 indicates that a Poor response condition is more likely to occur when the
stressor is Poor. For example, when the RR is 2.0, the chance that a lake is in Poor biological
(response) condition is twice as likely when the stressor is Poor than when the stressor is Not
Poor. Further details of RR and its interpretation, including estimation of a confidence interval
for RRest, can be found in Van Sickle et al. (2006).
10.4.4 Attributable risk
Population attributable risk (AR) measures what percent of the extent in Poor condition for a
biological response variable can be attributed causally to the Poor condition of a specific
stressor. AR is based on a scenario in which the stressor in Poor would be entirely eliminated
from the population of lakes, e.g., by means of restoration activities. That is, all lakes in Poor
condition for the stressor are restored to the Not Poor condition. AR is defined as the
proportional decrease in the extent of Poor biological response condition that would occur if
the stressor were eliminated from the population of lakes. Mathematically, AR is defined as
(Van Sickle and Paulsen (2008))
Pr(B = P) - Pr(B = P\S = NP)
AR~ Pr(B = P)
We estimated AR as
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BEPV — c/(a + c)
ARest = ' -
est BEPp
where
(c + d)
BEPp = - ^
(a + b + c + d)
and is the estimated proportion of the biological response that is in Poor condition. We
calculated a confidence interval for ARest following Van Sickle and Paulsen (2008).
An AR can take a value between 0 and 1. A value of 0 indicates either "No association" between
stressor and response, or else a stressor has a zero extent, i.e., is not present in the population.
A strict interpretation of AR in terms of stressor elimination, as described above, requires one
to assume that the stressor-response relation is strongly causal and that stressor effects are
reversible. Van Sickle and Paulsen (2008) discuss the reality of these assumptions, along with
other issues such as interpreting them when multiple, correlated stressors are present, and
using them to express the joint effects of multiple stressors.
However, AR can also be interpreted more informally, as a measure that combines RR and SE
into a single index of the overall, population-level impact of a stressor on a response. Van Sickle
and Paulsen (2008) show that the population attributable risk can be written as
SEPJRR - 1)
AD = 1
1 +SEPp(RR - 1)
This shows that the numerator of AR is the product of the SE of Poor stressor condition and the
"excess" RR, i.e., RR-1, of that stressor. The denominator standardizes this product to yield AR
values between 0 and 1. Thus, a high AR for a stressor indicates that the stressor is widely
prevalent (has a high SE of Poor condition), and the stressor also has a large effect (high RR) in
those lakes where it does have Poor condition.
10.4.5 Considerations when calculating and interpreting relative risk and attributable
risk
It is important to understand that contingency tables are created using a categorical, two-by-
two matrix; therefore, only two condition classes / stress levels can be used. There are three
ways in which condition classes / stress levels can be used for contingency tables:
• Good vs. Poor
• Good vs. Not-Good
• Not-Poor vs. Poor
where, "Not Good" combines fair and poor condition classes, and "Not Poor" combines good
and fair condition classes. In the first bulleted method, "Good vs. Poor" data associated with
the fair condition class is excluded from the analysis. Therefore, the results of the associated
calculation of relative risk are affected by which one of the above combinations is used to make
the contingency tables, and it is crucial that the objectives of the analysis are carefully
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considered to help guide this decision. For the NLA, for non-biological condition indicators (e.g.,
nutrients, physical habitat, etc.), a condition / stressor-level contingency table was created,
comparing the Not Poor condition class (i.e., a combination of good condition and fair
condition) to Poor condition class. This decision was made to indicate which stressors policy
makers and managers may want to prioritize for management efforts to improve poor
condition. After creating contingency tables, relative risk for each indicator was calculated.
A second consideration is that relative risk does not model joint effects of correlated stressors.
In other words, each stressor is modeled individually, when in reality, stressors may interact
with one another potentially increasing or decreasing impact on condition. This is an important
consideration when interpreting the results associated with relative risk.
To appropriately interpret attributable risk, it is important to understand that attributable risk
is associated with the following three major assumptions:
• Causality, or that the stressor causes an increased probability of poor condition;
• Reversibility, or that if the stressor is eliminated, causal effects will also be eliminated; and,
• Independence, or that stressors are independent of each other, so that individual stressor
effects can be estimated in isolation from other stressors.
These assumptions should be kept in mind when applying these results to management
decisions.
Attributable risk provides much needed insight into how to prioritize management for the
improvement of our aquatic ecosystems - lakes, in the case of the NLA. While the results of
attributable risk estimates are presented as percent area in poor condition that could be
reduced if the effects of a particular stressor were eliminated, these estimates are meant to
serve as general guidance as to what stressors are affecting condition and to what degree
(relative to the other stressors evaluated).
10.5 Change analysis
10.5.1 Background information
One of the objectives of the National Lakes Assessment (NLA) is to track changes over time. The
NLA conducted in 2022 was the fourth statistically valid survey of the nation's lakes and
reservoirs. In NLA 2007, lakes 4 hectares and larger were sampled. As discussed earlier in this
document, the NLA 2012 expanded the target population to include lakes within a smaller size
class category (1-4 hectares) and this remained the same for all subsequent surveys. Because of
this change in design, the change analysis was conducted on both the larger lakes (> 4 hectares)
and all lakes (>1 hectare) study populations.
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10.5.2 Data preparation
Analyses focused on the change in condition from the prior survey (2017) and the longest
duration for each study population. For the larger lakes study population, this included change
between 2007-2022 and 2017-2022. The larger lakes analyses included all sites from NLA 2007
(1130 sites), 801 NLA 2017 sites, and 775 NLA 2022 sites (NLA 2017 and 2022 excluded 1-4
hectare lakes). For the all lakes study population, change analyses included 2012-2022 and
2017-2022 and used data collected from all lakes sampled in 2012, 2017 and 2022.
10.5.3 Methods
Change analysis was conducted using the spsurvey package in R (Dumelle et al. 2023). Within
the GRTS (Generalized Random Tessellation Stratified) survey design, change analysis can be
conducted on continuous or categorical response variables (e.g., good, fair, and poor). The
analysis measures the difference between response variables of two survey time periods. For
NLA 2022, the categorical response variables were used to compare changes between NLA
2007 and 2012, 2012 and 2017, and 2017 and 2022. When using categorical response variables,
change is estimated by the difference in category estimates from the two surveys. Category
estimates are defined as the estimated proportion of values in each category, for example
good, fair, and poor.
Change between the two years is identified as statistically significant in the interactive data
dashboard and web-report when the resulting error bars around the change estimate do not
cross zero. Statistical significance is provided as a way to highlight results that may warrant
additional exploration and analyses.
For some indicators and subpopulations, the change in the percentage of lakes that is "not
assessed" can be relatively large and may change from survey to survey. Large changes in not
assessed may reflect changes in sampling or assessment success rather than actual changes in
condition associated with other categories such as good, fair and poor. Therefore, when the
percent of not assessed increases or decreases by more than 5 percentage points between
survey cycles, EPA will not present these results in the interactive dashboard to limit potentially
erroneous interpretations of condition change.
Change estimates could not be made for some indicators and some survey cycles due to
differences in methodologies (e.g., zooplankton), condition categories (i.e., lake drawdown),
and the timing of when indicators were added to the survey (e.g., atrazine added 2012,
cylindrospermopsin added 2017 and enterococci added 2022).
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10.6 Literature cited
Dumelle, M. T. Kincaid, A. R. Olsen, and M. Weber. 2023. Spsurvey: Spatial sampling design and
analysis in R. Journal of Statistical Software 105 (3): doi: 10.18637/jss.vl05.i03.
Olsen, A. R., and D. V. Peck. 2008. Survey design and extent estimates for the Wadeable
Streams Assessment. Journal of the North American Benthological Society 27:822-836.
Stevens, D. L., Jr, and S. F. Jensen. 2007. Sampling design, implementation, and analysis for
wetland assessment. Wetlands 27:515-523.
Stevens, D. L., Jr, and A. R. Olsen. 1999. Spatially restricted surveys over time for aquatic
resources. Journal of Agricultural, Biological, and Environmental Statistics 4:415-428.
Stevens, D. L., Jr, and A. R. Olsen. 2000. Spatially restricted random sampling designs for design-
based and model based estimation. Pages 609-616 in Accuracy 2000: Proceedings of the
4th International Symposium on Spatial Accuracy Assessment in Natural Resources and
Environmental Sciences. Delft University Press, The Netherlands.
Stevens, D. L., Jr, and A. R. Olsen. 2004. Spatially-balanced sampling of natural resources.
Journal of American Statistical Association 99:262-278.
Van Sickle, J., J. L. Stoddard, S. G. Paulsen, and A. R. Olsen. 2006. Using relative risk to compare
the effects of aquatic stressors at a regional scale. Environmental Management 38:1020-
1030.
Van Sickle, J., and S. G. Paulsen. 2008. Assessing the attributable risks, relative risks, and
regional extents of aquatic stressors. Journal of the North American Benthological
Society 27:920-931.
USEPA. 2006. Wadeable Streams Assessment: A Collaborative Survey of the Nation's Streams.
US Environmental Protection Agency, Office of Water and Office of Research and
Development, Washington, DC.
USEPA. 2009. National Lakes Assessment: A Collaborative Survey of the Nation's Lakes. US
Environmental Protection Agency, Office of Water and Office of Research and
Development, Washington, DC.
Whittier, T.R., Larsen, D.P., Peterson, S.A., and Kinkaid, T.M. 2002. A comparison of
impoundments and natural drainage lakes in the Northeast USA. Hydrobiologia 470:
157-171, doi.org/10.1023/A: 1015688407915
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Chapter 11: Quality Assurance Summary
The NLA has been designed as a statistically valid report on the condition of the Nation's lakes
at multiple scales, e.g., ecoregion (Level III and the aggregated nine NARS ecoregions). and
national, employing a randomized site selection process. The NLA is an extension of the EMAP
methods for assessing lakes, similar to the 1997 Northeastern Lakes Assessment; therefore, it
uses similar EMAP-documented and tested field methods for site assessment and sample
collection as the Northeast Lakes Assessment.
Key elements of the NLA Quality Assurance (QA) program include:
Quality Assurance Project Plan - A Quality Assurance Project Plan (QAPP) was developed and
approved by a QA team consisting of staff from the EPA's Office and Wetlands, Oceans and
Watersheds (OWOW) and Office of Environmental Information (OEI) and a Project QA Officer.
All survey participants signed an agreement to follow the QAPP standards. Compliance with the
QAPP was assessed through standardized field crew training and field crew assistance visits.
The QAPP addresses all aspects of the survey, including: project planning and management;
data quality objectives; sampling design and site selection; indicators; field crew assistance
visits; standardized/centralized data management; and data analysis. Detailed information on
site selection, field protocols and the laboratory sample processing are found in the following
documents:
• NLA 2022 SEG (EPA 841-B-21-008) - outlines the process to determine if a lake
meets the criteria for inclusion in the target population and is accessible for
sampling, and the appropriate replacement process if a lake is not sampleable;
• NLA 2022 FOM (EPA 841-B-21-011) - describes all field activites and protocols; and
• NLA 2022 LOM (EPA 841-B-21-010) - documentation of all laboratory methods.
Field Training and Sample Collection - EPA staff and contractors provided hybrid training that
included the review of online videos and quizzes and in person training sessions throughout the
study area. All field crew leads were required to complete all components of the NLA training
and field crew members were encouraged complete as much training as possible. All field crews
received an onsite assistance visit from a trained EPA staff member or contractor within the
first few weeks of fieldwork. Adjustments and corrections were made on the spot for any
problems identified during the assistance visit. To assure consistency, EPA supplied standard
sample/data collection equipment, sample bottles, filtration supplies, and shipping supplies for
all sampling events.
Revisits of Selected Field Sites - To evaluate within-year sampling variability, the NLA design
called for crews to revisit 10 percent of the sites selected in the design. These sites were
sampled twice in the NLA index period during a single year (visit 1 and visit 2). Useful metrics
and indicators tend to have high repeatability, that is among site variability will be greater than
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sampling variability based on repeat sampling at a subset of sites. To quantify repeatability,
NARS uses one of two metrics 1) Signal:Noise (S:N), or the ratio of variance associated with
sampling site (signal) to the variance associated with repeated visits to the same site (noise)
(Kaufmann et al., 1999) or 2) condition category contingency tables. When calculating S:N, all
sites are included in the signal, whereas only revisit sites contribute to the noise component.
Metrics with high S:N are more likely to show consistent responses to human caused
disturbance, and S:N values < 1 indicate that sampling a site twice yields as much or more
metric variability as sampling two different sites (Stoddard et al., 2008). The S:N values were
used by analysts in the process of selecting metrics and evaluating indicators.
Contingency tables are also used to visualize agreement between condition categories for the
first and second visits. These are presented for the NLA risk indicators that track detection and
risk (atrazine, cyanotoxins and enterococci).
Chemical Analyses - NLA 2022 used two labs for the water chemistry samples, the Wisconsin
State Lab of Hygiene (WSLOH; Wisconsin probability and state intensification sites) and the
Willamette Research Station (WRS; all remaining NLA and NES sites). For quality assurance of
chemical analyses, laboratories used QC samples which are similar in composition to samples
being measured. They provide estimates of precision and bias that are applicable to sample
measurements. To ensure the ongoing quality of data during analyses, every batch of water
samples was required to include QA samples to verify the precision and accuracy of the
equipment, reagent quality, and other quality measures. These checks were completed by
analyzing blanks or samples spiked with known or unknown quantities of reference materials,
duplicate analyses of the same samples, blank analyses, or other appropriate evaluations. The
laboratories reported QA results along with each batch of sample results. In addition,
laboratories reported holding times. Holding time requirements for analyses ensure analytical
results are representative of conditions at the time of sampling. To identify samples for
additional investigation, EPA reviewed all laboratory QA flags, data completeness, sample ionic
strength balance, completed several cross variably validity checks and noted any quality
failures.
For the atrazine samples, the NLA 2022 used two labs: WSLOH (Wisconsin sites) and
EnviroScience Inc. (ESCI), a sub-contracted laboratory with Great Lakes Environmental Cetner,
Inc. (all remaining sites). For the cyanotoxins samples, NLA 2022 used three labs including
WSLOH (Wisconsin sites), the EPA Region 4 Lab (R4 handpicked NES sites) and GreenWater
Laboratories (GWL), a sub-contracted laboratory with Avanti Corporation (all remaining NLA
sites). Proficiency test (PT) samples (5 concentrations per set and parameter) were sent to all
labs that analysed samples for atrazine and cyanotoxins (WSLOH - 1 set; EPA R4 1 set; GWL - 2
sets). The results from these tests were used to identify samples of acceptable quality for use in
the NLA assessment.
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Zooplankton Laboratory QA - EPA contracted with one lab for zooplankton sample processing.
This lab demonstrated that it could meet the QA/QC requirements identified in the NLA 2022
QAPP and LOM. These requirements included internal quality control (QC) checks on
zooplankton identification, the use of the Integrated Taxonomic Information System for
correctly naming species collected, and use of a standardized data management system.
Independent taxonomists were contracted to perform QC analysis of the primary lab's samples.
The external QC targeted the reidentification of 10% of the samples. The samples were
randomly selected. The reidentifications were made on new aliquots taken from the original
sample. Scheduling issues limited the processing of all samples selected, therefore only 7% (151
samples) were processed. Samples were assessed for within sample similarities using the Bray-
Curtis Dissimilarity Index (B - C) for each taxon. The zooplankton B-C data quality objective was
0.25 and the median B-C across all samples was 0.28. Although the median is slightly greater
than the DQO, all zooplankton samples were determined acceptable for further analysis since
the measure accounts for sample processing differences (each lab identified unique aliquots).
Benthic Macroinvertebrate Laboratory QA - NLA 2022 used one lab for benthic
macroinvertebrate sample processing. This lab demonstrated that it could meet the QA/QC
requirements identified in the NLA 2022 QAPP and LOM. These requirements included internal
quality control (QC) checks on sorting and identification of benthic macroinvertebrates and the
use of the Integrated Taxonomic Information System for correctly naming species collected, as
well as the use of a standardized data management system. Independent taxonomists were
contracted to perform QC analysis of 10% of the national lab samples. The QC samples were
randomly selected. Reidentifications were made with the same specimens (vials and slides of
individual were shipped to the QC lab) for 109 (10.1%) benthic samples. The mean percent
taxonomic disagreement (PTD) for the overall NLA 2022 benthic dataset was 9.3%, which is
better than the programmatic measurement quality objective of 15%.
Entry of Field Data and Quality Checks- NLA used a standardized data management structure,
1.e., use of the same standard field forms for data collected and centralized data management.
Most field data were collected electronically using an iPad with the NLA field data mobile
application. Following a review for accuracy and completeness, field crews submitted the
electronic forms directly from the NLA App to NARS IM, which automated upload to the NLA
2022 SQL database. No paper field forms were submitted in the 2022 survey.
Quality of field data were reviewed on a weekly, monthly and end of season basis using
numerous automated data quality checks. EPA staff and contractors then compiled a summary
of data quality issues which were sent to respective field crews to correct or provide additional
comments. If field data could not be corrected, crews were instructed to provide a comment as
to why field data could not be collected or measured. Corrected data and new comments were
resubmitted from the NLA App and updated in the NARS IM NLA 2022 SQL database.
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Data Management - Information management (IM) is integral to all aspects of the program
from initial selection of sampling sites through dissemination and reporting of final, validated
data. Quality measures implemented for the IM system are aimed at preventing corruption of
data at the time of their initial incorporation into the system and maintaining the integrity of
data and information after incorporation into the system.
Reconnaissance, field observation and laboratory analysis data were transferred from NLA
survey participants and collected and managed by the NARS IM center. Data and information
were managed using a tiered approach. First, all data transferred from a field team or
laboratory were physically organized (e.g., system folders) and stored in their original state.
Next, NARS IM created a synthesized and standardized version of the data to populate a
database that represented the primary source for all subsequent data requests, uses and
needs. All samples were tracked from collection to the laboratory.
The IM staff applied an iterative process in reviewing the database for completeness,
transcription errors, formatting compatibility, consistency issues and other quality control-
related topics. This first-line data review was performed primarily by NARS IM in consultation
with the NLA QA team. A second-phase data quality review consisted of evaluating the quality
of data based on MQOs as described in the QAPP. This QA review was performed by the NLA
QA team using a variety of qualitative and quantitative analytical and visualization approaches.
Records Management - EPA organizes and maintains all records associated with the survey.
Examples of the records include: all planning documents, such as the survey design, NLA QAPP,
SEG, FOM, LOM and other laboratory SOPs; QA implementation documents (e.g., QAPP
signature pages, crew training, assistance visit forms, lab verification information); data and
assessment files, draft reports and comments received. All data will eventually be archived in
the water quality portal.
Main Report - The main report provides a summary of the findings of each of the data analyses
and EPA's interpretation of them. The main report was extensively reviewed in-house by the
NLA team, its partners, and other EPA experts. Because previous reports using the same
analytical procedures were reviewed through an Independent External Review process, it was
determined that a letter review was not required for the main report. EPA used the comments
from the states and EPA's Office of Research and Development to refine the main report and
improve the clarity of documentation in this document.
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Appendix A: Lake Physical Habitat Expected Condition Models
Table 3 from TSD Chapter. Summary of regression models used in estimating lake-specific
expected values of Lake Physical Habitat variables RVegQx, LitCvrQx and LitRipCvrQx under least
disturbed conditions. Variable definitions and model details on following pages.
REGION v = RVeaQ
v = LitCvrQ
v = LitRipCvrQ
NAP Ly* =f(Lat, Lon, LkOrig, RDisIX,) Ly =f(L_LkArea, RDisIX)
(R2=23%, RMSE=0.162L*
(R2= 12% RMSE=0.281L)
Ly =f(Lat, Lon, LkOrig, RDisIX)
(R2=24%, RMSE=0.168L)
SAP Ly =f(Lon)
(R2=16%, RMSE=0.119L)
Ly =f(ElevXLon, RDisIX)
(R2=19%, RMSE=0.267L)
Ly =f(Lon, ElevXLon, Elev)
(R2=31%, RMSE= 0.148L)
CPL y =f(ElevXLat, RDisIX)
(R2=39%, RMSE=0 .0896)
y =f(L_Elev, RDisIX)
(R2=25%, RMSE= 0.174)
y =f( L_Elev, RDisIX)
(R2=44%, RMSE=0.093)
UMW Ly = (mean LRVegQ)
(R2=0%, RMSE=0.153L)
Ly = (mean LitCvrQ)
(R2=0%, RMSE=0.199L)
Ly = (mean LitRipCvrQ)
(R2=0%, RMSE=0 .115L)
CENPL Ly =f(hiiAg)
(R2=15%, RMSE=0.318L)
Ly =f(LkOrig, hiiAg)
(R2=9%, RMSE=0.276L)
Ly =f(hiiAg)
(R2=15%, RMSE=0.233L)
WMT Ly =f{Lat, Elev, L_LkArea, LkOrigin) Ly =f(Lat, Elev, L_LkArea,
(R2=28%, RMSE=0.167L) (R2=16%, RMSE=0.244L)
LkOrigin) Ly =f(Lat, Elev, L_LkArea, LkOrigin)
(R2=29%, RMSE=0.145L)
XER Ly =f(Lat, Elev)
(R2=24%, RMSE=0.284L)
Ly =f(Lat, Elev)
(R2=16%, RMSE=0.290L)
Ly =f( Lat, Elev)
(R2=21%, RMSE=0.265L)
*Ly refers to Logio-transformed lake habitat metric values.
**L refers to RMSE's that are in Logio units (e.g., 0.162L)
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VARIABLE DEFINITIONS
On following pages variables are defined as follows:
REF_NLA12 = Variable for disturbance level at site based on screening criteria from 2012, valid
values of L (least disturbed), I (intermediate disturbance), M (most disturbed), and ? (unknown
due to missing information)
REF_NLA17 = Variable for disturbance level at site based on screening criteria update for NLA
2017, valid values of L (least disturbed), I (intermediate disturbance), M (most disturbed), and ?
(unknown due to missing information)
Observed Habitat Indicator values are: (in the TSD text, these are abbreviated as RVegQ,
LitCvrQ. and LitRipCvrQ)
RVegQcl5, LitCvrQcl5, LitRipCvrQclS
L_RVegQcl5 = Log10(RVegQcl5 +0.01)
L_LitCvrQcl5 = Logio (LitCvrQcl5 +0.01)
L_LitRipCvrQcl5 = Logio (LitRipCvrQclS +0.01)
Expected Condition Regression Models have the form (in the TSD text. Expected condition
variables are abbreviated as RVegQX, LitCvrQX, and LitRipCvrQX):
L_RVegQc3xl5 = f(predictors) or RVegQc3xl5 = f(predictors)
L_LitCvrQc3xl5 = f(predictors) or LitCvrQc3xl5 = f(predictors)
L_LitRipCvrQc3xl5 = f(predictors) or LitRipCvrQc3xl5 = f(predictors)
Observed/Expected Condition Variables are defined as follows (in the TSD text. O/E variables
are abbreviated as RVegQ OE, LitCvrQ OE, and LitRipCvrQ OE)\
RVegQc30E15= (RVegQcl5/RVegQc3xl5) and Ll_RVegQc30E15 = Log10(RVegQc3OE15 +0.1)
LitCvrQc30E15= (LitCvrQcl5/LitCvrQc3xl5) and Ll_LitCvrQc30E15 = Logi0(LitCvrQc3OE15
+0.1)
LitRipCvrQc30E15= (LitRipCvrQcl5/LitRipCvrQc3xl5) and Ll_LitRipCvrQc30E15 =
Logio (LitRipCvrQc30E15 +0.1)
Predictors defined from variables in prk datafile NLA12 pc.nla lakeinfo all 20150415 are as
follows:
LATdd_use = LAT_DD_N83 = latitude in decimal degrees
LONdd_use = LON_DD_N83 = longitude in decimal degrees
ELEV_use = ELEVATION = lake surface elevation (meters above mean sea level)
L_ELEV_use = Log10(ELEV_use)
LkArea_km2 = LAKEAREA = lake surface area (km2)
L_LkAreakm2 = Logio(LkArea_km2)
Lake_Origin_use = LAKE_ORIGIN (with values: 'NATURAL' or 'MAN-MADE')
Reservoir = an indicator variable of Lake Origin, where
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If Lake_Origin_use = 'MAN-MADE' then Reservoir= 1;
If Lake_Origin_use = 'NATURAL' then Reservoir= 0;
Field human disturbance variables;
RDis_IX— index of near-shore human disturbance intensity and extent (see TSD text equation
5)
hiiAg proximity-weighted mean tally of up to 3 near-shore agricultural disturbances
(mean among stations
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NAP Expected PHab Reference Condition Models:
L_RVegQc3xl5 = 2.34593-(0.03705* LATdd_use)+(0.01723*LONdd_use)-(0.07954* Reservoir)
-(0.31865 *RDisJX);
Note: Reservoir = 0 for natural lakes, 1 for man-made reservoirs.
Rsq=0.2331 RMSE=0.16177 p<.0001 n=166/170;
Sites: All non-overlapping 2007-2012 NAP REF_NLA12 = L or I;
Set RDis_IX to zero (14% of 2007-&12 NAP sample sites have RDis_IX=0);
RVegQc3xl5=10**(L_RVegQc3xl5)-0.01;
Applied simple dirty models for LitCvr and LitRipCvr (see powerpoint file of regressions
6/13/14) that better define the influence of lake area — but then MUST include RDisJX,
because it is the strongest predictor of any of the 3 PHab indices if RT_NLA12_2015 S or T sites
are included with reference (R) sites;
Adjustment for reference distribution of O/E values:
L_RVegQc30E15= +0.04276 - (0.29150 RDisJX);
Rsq= 0.2026 RMSE=0.14469 p<0.0001 n=166/170;
Sites: All non-overlapping 2007-2012 NAP REF_NLA12 = L or I;
Ref O/E distribution based on Y-intercept of adjustment regression, but SD of ref sites only (not
S sites)
L_LitCvrQc3xl5= -0.8598 -(0.08109*L_LkAreakm2) - (0.28562*RDisJX);
Rsq=0.1228 RMSE=0.2808 p<0.0001 n=166/170;
Set RDis_IX to zero (14% of 2007-2012 NAP sample sites have RDis_IX=0);
Sites: All non-overlapping 2007-2012 NAP REF_NLA17 = L or I;
LitCvrQc3xl5= 10 * *(L_LitCvrQc3xl5)-0.01;
Adjustment for reference distribution of O/E values:
L_LitCvrQc3OE15= +0.04665 - (0.28240 RDisJX);
Rsq= 0.0592 RMSE=0.26819 p=0.0009 n=166/170;
Sites: All non-overlapping 2007-2012 NAP REF_NLA12 = L or I;
Ref O/E distribution based on Y-intercept of adjustment regression, but SD of ref sites only (not
S sites)
L_ Lit Rip CvrQc3xl 5= 2A1606-(0.03964*LATdd_use)+(0.01798*LONdd_use) -(0.08301*
Reservoir)
-{0.34039* RDisJX);
Note: Reservoir = 0 for natural lakes, 1 for man-made reservoirs.
Rsq=0.2407 RMSE=0.16783 p<0.0001 n=166/170;
Set RDisJX to zero (14% of 2007-2012 NAP sample sites have RDis_IX=0);
Sites: All non-overlapping 2007-2012 NAP REF_NLA17 = L or I;
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LitRip CvrQc3xl 5=10**(L_ LitRipCvrQc3xl5)-0.01;
Adjustment for reference distribution of O/E values:
L_LitRipCvrQc30E15= +0.04230 - (0.31323 RDisJX);
Rsq= 0.2075 RMSE=0.15095 p<0.0001 n=166/170;
Sites: All non-overlapping 2007-2012 NAP REF_NLA12 = Lor I;
Ref O/E distribution based on Y-intercept of adjustment regression, but SD of ref sites only (not
S sites).
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SAP -- Expected PHab Condition Models:
L_RVegQc3xl5= 0.24710 +(0.01012*LONdd_use);
Rsq=0.1637 RMSE=0.11878 p=0.0240 n=31/31;
Sites: AN non-ovelapping 2007-2012 SAP REF_NLA17 = L;
RVegQc3xl5= 10 * * (L_RVegQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_LitCvrQc3xl5= -0.66613 -(0.00000410*ElevXLon_use) -(0.51350*RDis_IX),
Rsq=0,1942 RMSE=0.26697 p=0.0487 n=31/31;
Set RDis_IX to zero (2% of 2007-2012 SAP sample sites have RDis_IX=0);
Sites: All non-overlapping 2007-2012 SAP REF_NLA17 = L;
LitCvrQc3xl5= 10 * * (L_LitCvrQc3xl5)-0.01;
Adjustment for reference distribution of O/E values:
L_LitCvrQcSOE15= +0.04287 - (0.46211 RDisJX);
Rsq= 0.0790 RMSE=0.24397 p=0.1255 n=31/31;
Sites: All non-overlapping 2007-2012 SAP REF_NLA12 = L;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
L_LitRipCvrQc3xl5=1.92708 -(0.000115130*ElevXLon_use) + (0.03141* LONdd_use) -
(0.00923 *ELEV_use);
Rsq=0.3083 RMSE=0.14817 p=0.0175 n=31/31;
Sites: All non-overlapping 2007-2012 SAP REF_NLA17 = L;
LitRip CvrQcSxl 5=10* *(L_ LitRipCvrQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
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CPL Expected PHab Condition Models:
RVegQc3xl5=0.35438 -0.00003019(ElevXLat_use) - 0.15193(RDisJX);
Rsq= 0.3868 RMSE=0.08963 p<0.0001 n=28/28;
Sites: All non-overlapping 2007-2012 CPL REF_NLA17 = L;
Set RDis_IX to lowest value in the region (4.4% have RDis_IX=0 in CPL);
Adjustment for reference distribution of O/E values:
L_RVegQc30E15= -0.0006653 - (0.22746 RDisJX);
Rsq= 0.0235 RMSE=0.21279 p=0.4362 n=28/28;
Sites: All non-overlapping 2007-2012 CPL REF_NLA12 = L;
Note: Regression keeping one low outlier with very little leverage;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
LitCvrQc3xl5= 0.71804 - (0.19300*L_Elev_use) - (0.12565*RDisJX);
Rsq= 0.2526 RMSE=0.17393 p<0.0001 n=28/28;
Sites: All non-overlapping 2007-2012 CPL REF_NLA17 = L;
Set RDisJX to lowest value in the region (0 in CPL);
Adjustment for reference distribution of O/E values:
LJitCvrQc30E15= -0.00743 - (0.09579 RDisJX);
Rsq= 0.0051 RMSE=0.1940 p=0.7178 n=28/28;
Sites: All non-overlapping 2007-2012 CPL REF_NLA12 = L;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
Lit Rip CvrQc3xl5= 0.59561 - (0.15322 *L_Elev_use) - (0.14358* RDisJX);
Rsq= 0.4423 RMSE=0.09293 p<0.0001 n=28/28;
Sites: All norepeat 2007-2012 CPL REF_NLA17 = L;
Set RDisJX to lowest value in the region (0 in CPL);
Adjustment for reference distribution of O/E values:
LJitRipCvrQc30E15= 0.01615 - (0.15265 RDisJX);
Rsq= 0.0312 RMSE=0.1234 p=0.3685 n=28/28;
Sites: All non-overlapping 2007-2012 CPL REF_NLA12 = L;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
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UMW Expected PHab Condition Models:
L_RVegQc3xl5= -0.61298;
****Dropped LON and LkArea - USED geometric (Log mean) NULL MODEL;
Rsq=0 RMSE=0.15333 n=49/50 ;
Sites: All non-overlapping 2007-2012 UMW REF_NLA17 = L;
RVegQc3xl5= 10 * * (L_RVegQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_LitCvrQc3xl5= -0.87559;
****Dropped survey year -- USED geometric (Log mean) NULL MODEL;
Rsq=0 RMSE=0.19944 p=N/A n=49/50;
Sites: All non-overlapping 2007-2012 UMW REF_NLA17 = L;
LitCvrQc3xl5= 10 * * {L_LitCvrQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_ LitRip CvrQc3xl5=-0.70830;
***** Dropped Lake Area -- USED geometric (Log mean) NULL MODEL;
Rsq=0 RMSE=0.11487 p=N/A n=49/50;
Sites: All non-overlapping 2007-2012 UMW REF_NLA17 = L;
LitRipCvrQcSxl5= 10 * * (L_LitRipCvrQc3xl5) -0.01;
LitCvrQc3xl5= 10 * * (L_LitCvrQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
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CENPL fNPL + SPL + TPL) Expected PHab Condition Models:
L_RVegQc3xl5=-0.75460- (0.0.86385*hiiAg);
Rsq=0.1532 RMSE=0.3178 p<0.0009 n=69/71;
Sites: All non-overlapping 2007-2012 CENPL_2015 REF_NLA17 = L, Excluding KS-R02 SD-101 (Oahi
Res) which has inadequate no of transects, but Includes Mound City res KS-R02 with corrected Elevation;
Set hiiAg to lowest value in the region (0)
Note: 2007-2012 NLA sites in CENPL with hiiAg=0 in NPL(>25%) SPL(>50%) TPL(75%)
RVegQc3xl5=10**(L_RVegQc3xl5)-0.01;
Adjustment for reference distribution of O/E values:
L_RVegQc30E15= 0.04688 - (0.80799 hiiAg);
Rsq= 0.1571 RMSE=0.29278 p=0.0007 n=69/71;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
L_LitCvrQc3xl5= -1.03378 + 0.10822*Reservoir -(0.38197*hiiAg);
Note: Reservoir = 0 for natural lakes, 1 for man-made reservoirs.
Rsq=0.0855 RMSE= 0.27579 p<0.0572 n=69/71;
Sites: All non-overlapping 2007-2012 CENPL_2015 REF_NLA17 = L
Set hiiAg to lowest value in the region (0)
Note: 2007-2012 NLA sites in CENPL with hiiAg=0 in NPL(>25%) SPL(>50%) TPL(75%)
LitCvrQc3xl5= 10 * *(L_LitCvrQc3xl5)-0.01;
Adjustment for reference distribution of O/E values:
L_LitCvrQc3OE15= 0.02752 - (0.35038 hiiAg);
Rsq= 0.0359 RMSE=0.28386 p=0.1255 n=69/71;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
L_LitRipCvrQc3xl5=-0.82455-(0.61960* hiiAg);
Rsq=0.1471 RMSE=0.23336 p=0.0011 n=69/71;
Sites: All non-overlapping 2007-2012 CENPL_2015 REF_NLA17 = L
Set hiiAg to lowest value in the region (0)
Note: 2007-2012 NLA sites in CENPL with hiiAg=0 in NPL(>25%) SPL(>50%) TPL(75%)
Lit Rip CvrQc3xl 5=10* *(L_ LitRipCvrQc3xl5) -0.01;
Adjustment for reference distribution of O/E values:
L_LitRipCvrQc30E15= 0.04303 - (0.59485 hiiAg);
Rsq= 0.1465 RMSE=0.22462 p=0.0012 n=69/71;
Ref O/E distribution based on Y-intercept and RMSE of adjustment regression.
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**** Note: If remove sites East of approximately -95 degrees LON that removes all hiiAg so
association with LON is largely assoc with hiiAg - adopted conservative model without LON.
See dirty models for all three indices with hiiAg alone (prk 3/13/15 SAS EnterpriseGuide
projects) for all three of the above, they all have higher Rsq, similar RMSE, similar intercepts,
similar slopes pcO.OOGl n= 669/694 to 673/694.
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WMT Expected PHab Condition Models:
L_RVegQc3xl5= 0.53572-(0.00008953*£I£V_use)-
(0.25957* Reservoir)+(0.07296* L_LkAreakm2)
-(0.01939* LATdd_use);
Note: Reservoir = 0 for natural lakes, 1 for man-made reservoirs,
Rsq=0.2825 RMSE=0.16743 p=0.0001 n=74/75;
Sites: All non-overlapping 2007-2012 WMT REF_MLA17 = L;
RVegQc3xl5=10**(L_RVegQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_LitCvrQc3xl5=-1.10550-(0.00004299* ELEV_use)-
(0.05083* L_LkAreakm2)+(0.00407*LATdd_use)
-(0.18384* Reservoir);
Note: Reservoir = 0 for natural lakes, 1 for man-made reservoirs.
Rsq=0.1555 RMSE=0.24373 p=.0187 n=74/75;
Sites: All non-overlapping 2007-2012 WMT REF_NLA17 = L;
LitCvrQc3xl5= 10 * *{L_LitCvrQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_LitRipCvrQc3xl5= -0.08802-(0.00006666*ELEV_use)+(0.04200*L_LkAreakm2)-
(0.01015* LATdd_use)-(0.22650* Reservoir);
Note: Reservoir = 0 for natural iakes, 1 for man-made reservoirs.
Rsq=0.2922 RMSE=0.14513 p<.0001 n=74/75;
Sites: All no-repeat 2007-2012 WMT REF_NLA17 = L;
LitRip CvrQcSxl 5=10* *(L_ LitRipCvrQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
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XER Expected PHab Condition Models:
L_RVegQc3xl5= 0.44708 -(0.02612 *LATdd_use) -(0.00013249*ELEV_use);
Rsq=0.2365 RMSE=0.28355 p=0.1009 n=20/21;
Sites: All no-repeat 2007-2012 XER REF_NLA17 = L;
RVegQc3xl5=10**(L_RVegQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_LitCvrQc3xl5=0.08706-(0.02849* LATdd_use)-(0.00003932* ELEV_use);
Rsq=0.1578 RMSE=0.29004 p=0.2322 n=20/21;
Sites: All no-repeat 2007-2012 XER REF_NLA17 = L;
*** Note this was 8th best in All Subsets Regression models with <=2 predictors ranked by Cp;
*** Note this was 6th best in All Subsets ranked by Rsq;
*** Consistent model across all the indicators and across full set of sites;
LitCvrQc3xl5= 10 * *(L_LitCvrQc3xl5)-0.01;
Ref O/E distribution based on mean and SD of ref sites.
L_ Lit Rip CvrQc3xl5=0.24931 - (0.02529* LATdd_use)-(0.00010090* ELEV_use);
Rsq=0.2115 RMSE= 0.26455 p=0.1327 n=20/21;
Sites: All no-repeat 2007-2012 XER REF_NLA17 = L;
Lit Rip CvrQc3xl 5=10* *(L_ LitRipCvrQc3xl5) -0.01;
Ref O/E distribution based on mean and SD of ref sites.
NOTE 3/13/15 prk: Reexamined models. The p-values (and of course also r2 and RMSE) not
improved by using
single predictors (ELEV_use LATdd_use and ELEVxLatdd_use). The mechanisms and univariate
plots of these single predictors all convincing and support the 3 models above;
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NLA 07,12,17 — P.R.Kaufmann April 27, 2020
Log 10fObserved/Expected] Lake Habitat Cover & Structural Complexity
Versus Anthropogenic Disturbance Stress (LIM-2020) and Year
For 9 Ecoregions (Sample stats, not weighted -%iles: 5/25/50/75/95
w/outliers shown as"+"
Following figures present the O/E values vs LIM for
the three PHab indicators for the three surveys for
each of the 9 Ecoregions.
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Riparian Vegetation:
NAP
L I M L I M L I M
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Riparian Vegetation:
SAP
L I M L I M L I M
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Riparian Vegetation:
CPL
M
M
2017
$
—j—
M
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Riparian Vegetation:
UMW
L I M L I M L I M
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Riparian Vegetation:
TPL
o
+
LU
o
CO
o
O
CD
>
DC
O)
o
M
M
~r
L
M
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Riparian Vegetation:
NPL
1.0
0.5
o
+
LU
o
CO
o
O
CO
:>
CD
o
0.0
-0.5
-1.0
T
L
M
M
2017
$
-r-
L
M
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Riparian Vegetation:
SPL
L I M L I M L I M
175
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Riparian Vegetation:
WMT
L I M L I M L I M
176
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Riparian Vegetation:
XER
i 1 1— —i 1 1— —i 1 1-
L I M L I M L I M
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Littoral Habitat:
NAP
L I M L I M L I M
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Littoral Habitat:
SAP
L I M L I M L I M
179
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Littoral Habitat:
CPL
L 1 M L I M L I M
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Littoral Habitat:
UMW
L I M L I M L I M
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Littoral Habitat:
TP I
L I M L I M L I M
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Littoral Habitat:
NPL
2007
2012
2017
1.0
0.5'
O
+
LU
o
CO
o
O
>
O
0
0,0
CD
o
-0.5
-1.0'
"I-
L
M
M
M
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Littoral Habitat
SPL
L 1 M L I M L I M
184
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Littoral Habitat:
WMT
L I M L I M L I M
185
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Littoral Habitat:
XER
M
-| r
L I M
M
186
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Littoral-Riparian Complexity:
NAP
1.0-
0.5
0.0-
-0.5
-1.0-
2007
0n
M
M
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Littoral-Riparian Complexity:
SAP
LLI
O
CO
o
O
k_
>
O
Q.
hi
CD
o
M
M
M
188
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Littoral-Riparian Complexity:
CPL
2017
$
M
I T M L I M
189
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Littoral-Riparian Complexity:
UMW
o
+
LU
O
CO
o
O
>
O
Q.
Cd
CD
O
M
M
M
190
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Littoral-Riparian Complexity:
TPL
o
+
LU
O
CO
o
O
L_
>
o
Q.
ir
CD
O
M
M
M
191
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Littoral-Riparian Complexity:
NPL
o
+
LU
O
CO
y
O
L_
>
o
Q.
o>
o
M
M
M
192
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Littoral-Riparian Complexity:
SPL
~T~
L
M
M
T
L
M
193
-------
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Littoral-Riparian Complexity:
WMT
M
M
M
194
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Littoral-Riparian Complexity:
XER
o
+
LLI
o
CO
O
o
>
o
Q.
cc
CD
O
M
M
M
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Appendix B: Survey Design and Estimated Extent Summary for NLA 2007, 2012, 2017 and 2022
Category
Characteristic
Description
2022
2017
2012
2007
sampling frame
sampling frame
source
NHDPIus HR
NHDPIus and
NHDPIus HR for 1-
5 ha lakes
NHDPIus, version
2
NHD
sampling frame
sampling frame
total number of
6,512,454
586,678
378,858
389,005
lake objects in
(waterbody
(lake objects)
(lake objects)
(lake objects)
source (NHD)
polygons)
sampling frame
sampling frame
lake objects
included in the
sampling frame
497,840
465,901
277,886
123,369
sampling frame
sampling frame
exclusions
lake objects
excluded because
they are not
expected to meet
the target
population
definition
6,014,614
120,777
100,972
265,636 (of which
233,627 were 1-
4ha)
survey design
survey design
GRTS
GRTS stratified by
GRTS with
GRTS with
stratified by
state and unequal
stratification and
stratification and
state and
probability of
unequal
unequal
unequal
selection by lake
probability of
probability of
probability
size within state
selection by lake
selection by lake
by lake size
size within state
size within state
within state
survey design
restriction
minimum lakes
per state
8
7
7
7
survey design
restriction
maximum lakes
per state
50
50
43
none
survey design
stratification
stratification
by state
by state
by state and
NLA12_CLS
None
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Category
Characteristic
Description
2022
2017
2012
2007
survey design
lake area
categories
description (ha)
(1-4 ], (4-10],
(10-50], >50
(1-4 ], (4-10], (10-
20], (20-50], >50
(1-4], (4-10], (10-
20], (20-50], >50
(4-10], (10-20],
(20-50], (50-100],
>100
survey design
lake area
categories
minimum (ha)
1 ha
1 ha
1 ha
4 ha
survey design
expected unique
lakes
total lakes
904
904
904
909
survey design
expected sample
size
total visits
1,000
1,000
1,000
1,000
survey design
expected split
new/previously
sampled
50/50
50/50
62/38
NA
survey design
revisits
number of lakes
96
96
96
91
survey
implementation
survey design
lakes sampled
total lakes
samples (used in
population
estimates)
981
1,005
1,038
1130
survey
implementation
survey design
lakes sampled
Size class: 1-4 ha
216
204
87
0
survey
implementation
survey design
lakes sampled
Size class: 4-10 ha
195
179
142
73
survey
implementation
survey design
lakes sampled
Size class: 10-50
ha
293
NA
NA
NA
survey
implementation
survey design
lakes sampled
Size class: 10-20
ha
NA
192
173
162
survey
implementation
survey design
lakes sampled
Size class: 20-50
ha
NA
164
225
211
survey
implementation
survey design
lakes sampled
Size class: >50 ha
277
266
411
684
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Category
Characteristic
Description
2022
2017
2012
2007
estimated extent
estimated lake
population
target
population:
All lakes > lha
(LCB95Pct-
UCB95Pct)
268,018
(256,329-
279,706)
224,916
(194,076-
255,755)
126,113*
NA
estimated extent
estimated lake
population
target
population:
Large lakes > 4 ha
98,519
76,177
68,777
65,259*
estimated extent
estimated lake
population
target unknown
NA
3,290
3,538
—
estimated extent
estimated lake
population
non-target lakes
229,822
237,695
114,695
55,146
estimated extent
estimated lake
population
sampled
population
124,309
109,701
111,818
49,546
estimated extent
estimated lake
population
NLA report result
representation
target
population
target population
sampled
population
sampled
population
*Upper and lower confidence intervals (CI) are not provided since reporting changed from the sampled population to the target population
in 2017. Estimated target population values for 2012 and 2007 were updated in 2017. Weights for all survey years can be found in the "Data
for Population Estimate" files on the NARS Data page.
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Appendix C: NLA 2022 Indicator Benchmark Summary
Physical habitat benchmarks are not included since regionally relevant lake-specific benchmarks are modeled.
Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Coastal
Plains
Good
>51.8
Sample collected from the lake
bottom at 10 shoreline locations
and composited for each lake.
Organisms were usually identified
to genus and an index was
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Eastern
Highlands
Good
>44.5
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Plains
Good
>39.5
—
developed based on life history
characteristics and tolerance to
environmental conditions.
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Upper
Midwest
Good
>51.4
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Western
Mountains
Good
>47.6
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Coastal
Plains
Poor
<44.1
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Eastern
Highlands
Poor
<31.4
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Plains
Poor
<26.6
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Upper
Midwest
Poor
<37.2
—
Biological
Benthic
Macroinvertebrate
NLA-derived regionally specific
benchmark
Western
Mountains
Poor
<32.6
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Coastal
Plains
Good
>59.42
Sample collected from the water
column at the open-water site.
Organisms were usually identified
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Eastern
Highlands
Good
>73.595
—
to genus and an index was
developed based on life history
characteristics and tolerance to
environmental conditions.
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Plains
Good
>36.72
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Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Upper
Midwest
Good
>63.68
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Western
Mountains
Good
>60.78
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Coastal
Plains
Poor
<53.77
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Eastern
Highlands
Poor
<60.03
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Plains
Poor
<28.17
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Upper
Midwest
Poor
<52.03
—
Biological
Zooplankton
NLA-derived regionally specific
benchmark
Western
Mountains
Poor
<51.32
—
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
CPL
Good
<12.7
ug/L
Sample collected from a vertically
integrated water column at the
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
NAP
Good
<4.52
ug/L
open-water site. Measured
concentrations were compared to
benchmarks.
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
NPL
Good
<10.9
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
SAP
Good
<5.54
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
SPL-
manmade
Good
<8.97
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
SPL-natural
Good
<118
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
TPL
Good
<13.9
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
UMW
Good
<6.7
ug/L
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Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
WMT
Good
<1.83
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
XER
Good
<5.92
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
CPL
Poor
>28
ug/L
Sample collected from a vertically
integrated water column at the
open-water site. Measured
concentrations were compared to
benchmarks.
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
NAP
Poor
>8.43
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
NPL
Poor
>19.3
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
SAP
Poor
>13.1
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
SPL-
manmade
Poor
>12.6
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
SPL-natural
Poor
>219
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
Temperate
Plains
Poor
>19.8
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
UMW
Poor
>14.6
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
WMT
Poor
>3.86
ug/L
Biological
Chlorophyll a
NLA-derived regionally specific
benchmark
XER
Poor
>9
ug/L
Chemical
Acidity
Nationally consistent, literature
benchmark described in Herlihy
etal. (1991)
National
Good
ANC > 50
ueq/L
ueq/L
ANC (corrected for DOC) measured
from a vertically integrated water
column at the open-water site.
Chemical
Acidity
Nationally consistent, literature
benchmark described in Herlihy
etal. (1991)
National
Poor
ANC < 0
Ueq/L and
DOC values
< 6 mg/L
ueq/L
Measured concentrations were
compared to benchmarks.
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Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Chemical
Atrazine
EPA aquatic plant concentration
equivalent level of concern (CE-
LOC); click here
National
Above
Benchmark =
Poor
> 3.4 ppb
ppb
Sample collected from a vertically
integrated water column sample at
the open-water site. Measured
concentrations were compared to
benchmark.
Chemical
Atrazine
Atrazine minimum detection
level (MDL)
National
Detected
> or = 0.046
ppb
Chemical
**Cylindrospermopsin
EPA recreational water qualtiy
criteria and swimming advisory
recommendation. USEPA 2019.
EPA 822-R-19-001.
National
Above
Benchmark =
Poor
>15
ppb
Sample collected from a vertically
integrated water column sample at
the open-water site. Measured
concentrations were compared to
benchmark.
Chemical
**Cylindrospermopsin
Cylindrospermopsin minimum
detection level (MDL)
National
Detected
> or = 0.05
ppb
Chemical
Microcystins
EPA recreational water qualtiy
criteria and swimming advisory
recommendation. USEPA 2019.
EPA 822-R-19-001.
National
Above
Benchmark =
Poor
>8
ppb
Sample collected from a vertically
integrated water column sample at
the open-water site. Measured
concentrations were compared to
benchmark.
Chemical
Microcystins
Microcystin minimum detection
level (MDL)
National
Detected
> or = 0.1
ppb
Chemical
**Enterococci
EPA Statistical Threshold Value
USEPA 2012. EPA 820-F-12-058
National
Above
Benchmark =
Poor
>1,280
CCE/
100
mL
Sample collected from last littoral
station or the launch site in an area
that was approximately 1 m deep
at about 0.3 m (12 inches) below
the water.
Chemical
Oxygen (Dissolved)
Nationally consistent, literature
benchmark; warmwater daily
minimum for "other life stages";
US EPA 1986. Quality Criteria for
Water ("Gold Book")
National
Poor
<= 3 ppm
ppm
Measures were collected from the
in-situ oxygen measure from the
top 2m of the profile at the index
site. The mean of all measurements
between 0 and 2 meters was
compared to the benchmark.
Chemical
Oxygen (Dissolved)
Nationally consistent, literature
benchmark; warmwater daily
minimum for "early life stages";
US EPA 1986. Quality Criteria for
Water ("Gold Book")
National
Good
>= 5 ppm
ppm
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
CPL
Good
<659
ug/L
Sample collected from a vertically
integrated water column at the
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Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
NAP
Good
<428
ug/L
open-water site. Measured
concentrations were compared to
benchmarks.
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
NPL
Good
<849
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
SAP
Good
<266
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
SPL-
manmade
Good
<650
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
SPL-natural
Good
<7840
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
TPL
Good
<865
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
UMW
Good
<766
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
WMT
Good
<253
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
XER
Good
<605
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
CPL
Poor
>923
ug/L
Sample collected from a vertically
integrated water column at the
open-water site. Measured
concentrations were compared to
benchmarks.
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
NAP
Poor
>655
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
NPL
Poor
>1620
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
SAP
Poor
>409
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
SPL-
manmade
Poor
>830
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
SPL-natural
Poor
>11100
ug/L
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Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
TPL
Poor
>1350
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
UMW
Poor
>926
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
WMT
Poor
>429
ug/L
Chemical
Total Nitrogen
NLA-derived regionally specific
benchmark
XER
Poor
>954
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
CPL
Good
<43
ug/L
Sample collected from a vertically
integrated water column at the
open-water site. Measured
concentrations were compared to
benchmarks.
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
NAP
Good
<16
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
NPL
Good
<63
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
SAP
Good
<18
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
SPL-
manmade
Good
<30
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
SPL-natural
Good
<486
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
TPL
Good
<38.4
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
UMW
Good
<24.8
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
WMT
Good
<23.4
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
XER
Good
<44
ug/L
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Metric
Category
Indicator
Benchmark Description
National/
Ecoregion
Condition class
Value
Units
General Assessment Notes
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
CPL
Poor
>59.5
ug/L
Sample collected from a vertically
integrated water column at the
open-water site. Measured
concentrations were compared to
benchmarks.
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
NAP
Poor
>27.9
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
NPL
Poor
>82
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
SAP
Poor
>33
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
SPL-
manmade
Poor
>43
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
SPL-natural
Poor
>839
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
TPL
Poor
>57.5
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
UMW
Poor
>40
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
WMT
Poor
>43
ug/L
Chemical
Total Phosphorus
NLA-derived regionally specific
benchmark
XER
Poor
>84.8
ug/L
Chemical
Trophic State
Nationally consistent, NLA-
derived benchmark
National
Oligotrophic
<2
ug/L
Sample collected from a vertically
integrated water column at the
open-water site.
Trophic state was based on
measured chlorophyll a
concentrations.
Chemical
Trophic State
Nationally consistent, NLA-
derived benchmark
National
Mesotrophic
>2 and <7
ug/L
Chemical
Trophic State
Nationally consistent, NLA-
derived benchmark
National
Eutrophic
>7 and <30
ug/L
Chemical
Trophic State
Nationally consistent, NLA-
derived benchmark
National
Hypereutrophic
>30
ug/L
** identifies new or updated benchmarks for NLA 2022
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Appendix D: Zooplankton
11.1 List of candidate metrics for zooplankton
This section provides additional details for the candidate metrics we considered when developing the
MMIs for each bio-region. Tables D.l through D.5 list each metric by its variable name, which of the six
metric categories it was assigned to (see Section 7.4.3), and a description of the metric for the Coastal
Plains, Eastern Highlands, Plains, Upper Midwest, and Western Mountains bio-regions, respectively. In
addition, the responsiveness to disturbance and repeatability of each metric is provided (t-value for
responsiveness, ad S:N value for repeatability).
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Table D. 1. List of candidate metrics used to develop the zooplankton MMI for the Coastal Plains bioregion.
t value
(Least disturbed
Mean Value for
Mean Value for
vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Abundance/
Biomass of individuals of smaller-sized taxa
Biomass/
(NET_SIZECLS_NEW=FINE; coarse and fine
Density
FINE BIO
net samples combined)
14.7
50.2
-1,67
1.2
Abundance/
Biomass/
Biomass represented by individuals
Density
ZOFN BIO
collected in fine mesh net (50-um)
20.5
67.2
-1.79
1.2
Percent of total individuals that are within
the cladoceran family Sididae (coarse and
Cladoceran
SIDID PIND
fine net samples combined)
2.10
8.18
-1.80
0.4
Total density of individuals within the
copepod order Calanoida (coarse and fine
Copepod
CALAN DEN
net samples combined)
5.6
22.9
--1.30
1.9
Number of families represented by distinct
native taxa (coarse and fine net samples
Richness/Diversity
FAM NAT NTAX
combined)
11.9
9.3
2.66
1.9
Number of families represented by distinct
taxa (coarse and fine net samples
Richness/Diversity
FAM NTAX
combined)
11.9
9.4
2.55
2.0
Number of genera represented by distinct
taxa (coarse and fine net samples
Richness/Diversity
GEN NTAX
combined)
15.4
12.1
2.21
1.5
Number of genera represented by distinct
native taxa (coarse and fine net samples
Richness/Diversity
GEN NAT NTAX
combined)
15.3
11.9
2.29
1.3
Number of families represented by distinct
Richness/Diversity
ZOFN FAM NAT NTAX
native taxa in the fine mesh net (50-um)
7.4
5.4
2.32
1.4
Total density of individuals within the
rotifer order Collothecaceae (coarse and
Rotifer
COLLO BIO
fine net samples combined)
0.22
0.02
1.79
3.3
Percent of total individuals within the
rotifer order Collothecaceae (coarse and
Rotifer
COLLO PIND
fine net samples combined)
2.27
0.32
1.87
2.0
Percent of total biomass within the rotifer
order Collothecaceae (coarse and fine net
Rotifer
COLLO PBIO
samples combined)
1.0
0.15
1.8
7.2
Number of distinct predator taxa (coarse
Trophic
PRED NTAX
and fine net samples combined)
2.5
1.3
2.56
4.6
Percent of distinct taxa that are predators
Trophic
PRED_PTAX
(coarse and fine net samples combined)
12.01
6.59
2.71
2.2
207
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NLA 2022 Technical Support Document - August 2024
t value
(Least disturbed
Mean Value for
Mean Value for
vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Number of distinct herbivore taxa (coarse
Trophic
HERB NTAX
and fine net samples combined)
11.9
8.8
2.22
2.1
Percent of distinct taxa that are
omnivorous (coarse and fine net samples
Trophic
OMNI PTAX
combined)
22.03
34.10
-3.35
4.3
Percent of total density represented by
omnivorous individuals (coarse and fine
Trophic
OMNI PDEN
net samples combined)
18.12
39.82
-2.37
1.7
Number of distinct rotifer taxa that are
predators (coarse and fine net samples
Trophic
ROT PRED NTAX
combined)
2.2
1.1
2.50
4.5
Percent of distinct rotifer taxa that are
Trophic
ROT PRED PTAX
predators
10.78
5.64
2.70
1.9
Number of distinct rotifer taxa that are
herbivores (coarse and fine net samples
Trophic
ROT HERB NTAX
combined)
6.8
4.6
2.00
1.8
Biomass represented by rotifer individuals
Trophic
ROT OMNI BIO
that are omnivores
4.8
35.0
-1.76
1.4
Percent of rotifer individuals represented
Trophic
ROT OMNI PIND
by omnivores
13.41
26.55
-1.88
2.0
Percent of distinct rotifer taxa that are
Trophic
ROT OMNI PTAX
omnivorous
17.26
27.95
-3.34
2.6
Percent of rotifer density represented by
Trophic
ROT OMNI PDEN
omnivores
17.82
39.27
-2.36
1.7
Metrics Derived from 300-count Subsamples of Coarse and Fine Net Sa
mples
Abundance/
Biomass
Total biomass in 300-count subsample of
Density
ZOFN300 BIO
fine-mesh net sample (50-^m)
11.6
34.6
-1.73
1.0
Percent of distinct taxa in the 300-count
subsamples that are in the family
Bosminidae (coarse and fine net samples
Cladoceran
BOSM300 PTAX
combined)
7.98
4.07
2.94
0.3
Percent of individuals within the
cladoceran family Sididae in 300-count
subsamples (coarse and fine net samples
Cladoceran
SIDID300 PIND
combined)
2.95
9.10
-1.68
0.7
Percent of biomass in dominant copepod
taxon in the 300 count subsamples (coarse
Copepod
DOM1 300 COPE PBIO
and fine net samples combined)
85.21
79.61
0.86
1.9
Number of genera represented by distinct
taxa (coarse and fine net samples
Richness/Diversity
GEN300 NTAX
combined)
14.1
11.1
2.16
1.8
208
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t value
(Least disturbed
Mean Value for
Mean Value for
vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Number of genera represented by distinct
native taxa (coarse and fine net samples
Richness/Diversity
GEN300 NAT NTAX
combined)
14.1
11.0
2.24
1.5
Number of families represented in 300
count subsamples (coarse and fine net
Richness/Diversity
FAM300 NTAX
samples combined)
10.9
8.6
2.61
2.2
Number of native families represented in
300 count subsamples (coarse and fine net
Richness/Diversity
FAM300 NAT NTAX
samples combined)
10.9
8.6
2.72
2.1
Number of distinct native families in 300-
count subsample of fine-mesh net sample
Richness/Diversity
ZOFN300 FAM NAT NTAX
(50-nm)
6.7
4.8
2.49
1.4
Biomass represented by individuals of the
rotifer order Collothecaceae in the 300-
count subsamples (coarse and fine net
Rotifer
COLLO300 BIO
samples combined)
0.08
0.01
1.81
7.0
Percent of biomass within the rotifer order
Collothecaceae in the 300-count
subsamples (coarse and fine net samples
Rotifer
COLLO300 PBIO
combined)
0.96
0.16
1.75
5.9
Number of distinct taxa that are predators
in 300 count subsamples (coarse and fine
Trophic
PRED300 NTAX
net samples combined)
1.7
1.0
1.94
2.7
Biomass of predator individuals in 300
count subsamples (coarse and fine net
Trophic
PRED300 BIO
samples combined)
0.46
0.14
2.45
1.5
Number of distinct taxa that are herbivores
in 300 count subsamples (coarse and fine
Trophic
HERB300 NTAX
net samples combined)
10.9
7.8
2.58
1.8
Percent of omnivorous individuals in 300
count subsamples (coarse and fine net
Trophic
OMNI300 PIND
samples combined)
15.54
28.43
-1.85
1.4
Percent of distinct taxa that are omnivores
in 300 count subsamples (coarse and fine
Trophic
OMNI300 PTAX
net samples combined)
23.75
37.16
-2.91
4.1
Percent of biomass represented by
omnivorous individuals in 300 count
subsamples (coarse and fine net samples
Trophic
OMNI300 PBIO
combined)
27.14
33.99
-0.79
1.2
Number of distinct rotifer taxa that are
predators in 300 count subsamples (coarse
Trophic
ROT_PRED300_NTAX
and fine net samples combined)
1.7
1.0
1.940
2.7
209
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NLA 2022 Technical Support Document - August 2024
t value
(Least disturbed
Mean Value for
Mean Value for
vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Biomass represented by rotifer individuals
that are predators in 300 count
subsamples (coarse and fine net samples
Trophic
ROT
PRED300 BIO
combined)
0.46
0.14
2.45
1.5
Number of distinct rotifer taxa that are
herbivores in 300 count subsamples
Trophic
ROT
HERB300 NTAX
(coarse and fine net samples combined)
6.0
3.7
2.45
1.4
Percent of rotifer individuals that are
omnivorous in 300 count subsamples
Trophic
ROT
OMNI300 PIND
(coarse and fine net samples combined)
12.24
25.10
-2.00
1.9
Percent of distinct rotifer taxa that are
omnivorous in 300 count subsamples
Trophic
ROT
OMNI300 PTAX
(coarse and fine net samples combined)
18.35
30.18
-3.06
3.6
210
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NLA 2022 Technical Support Document - August 2024
Table D. 2. List of candidate metrics used to develop the zooplankton MMI for the Eastern Highlands bio-region
t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Abundance/
Density represented by individuals collected in
Biomass/
coarse mesh net (150-um for 2012 samples, 243 um
Density
ZOCN DEN
for 2007 resamples)
12.56848
34.33432549
-1.89
7.1
Abundance/
Density represented by native individuals collected
Biomass/
in coarse mesh net (150-um for 2012 samples, 243
Density
ZOCN NAT DEN
um for 2007 resamples)
12.56848
34.33106863
-1.89
2.1
Abundance/
Density represented by individuals of taxa collected
Biomass/
in coarse mesh net (150-um; coarse and fine net
Density
COARSE DEN
samples combined)
21.26666667
53.84573922
-2.13
2.4
Abundance/
Biomass represented by individuals of taxa
Biomass/
collected in coarse mesh net (150-um; coarse and
Density
COARSE PBIO
fine net samples combined)
68.49155556
56.48058824
1.86
1.7
Abundance/
Density represented by individuals of native larger-
Biomass/
sized taxa (NET_SIZECLS_NEW=COARSE; coarse and
Density
COARSE NAT DEN
fine net samples combined)
21.266666667
53.80877451
-2-12
1.5
Abundance/
Biomass represented by individuals of native larger-
Biomass/
sized taxa (NET_SIZECLS_NEW=COARSE; coarse and
Density
COARSE NAT PBIO
fine net samples combined)
68.491555556
56.44254902
1.86
1.5
Abundance/
Biomass represented by individuals of smaller-sized
Biomass/
taxa (NET_SIZECLS_NEW=FINE; coarse and fine net
Density
FINE PBIO
samples combined)
31.508444444
43.519411765
-1.86
1.7
Density of native individuals within the suborder
Cladoceran
CLAD DEN
Cladocera (coarse and fine net samples combined)
6.813766667
27.71694902
-1.94
1.9
Density of native individuals within the suborder
Cladoceran
CLAD NAT DEN
Cladocera (coarse and fine net samples combined)
6.813766667
27.71382549
-1.94
1.8
Biomass represented by large cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE; coarse and fine net
Cladoceran
LGCLAD BIO
samples combined)
25.780533111
10.663794725
2.16
1.3
Biomass represented by native large cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE; coarse and fine net
Cladoceran
LGCLAD NAT BIO
samples combined)
25.780533111
10.656975706
2.16
1.3
Biomass represented by small cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD BIO
samples combined)
2.985147667
31.80179637
-2.37
2.6
Density represented by small cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCERAN_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD_DEN
samples combined)
2.476364444
22.86743922
-1.99
2.4
211
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NLA 2022 Technical Support Document - August 2024
t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of small cladoceran individuals
(SUBORDER=CLADOCERA and CLAD-SIZE=SMALL;
Cladoceran
SMCLAD PIND
coarse and fine net samples combined)
9.58
17.42
-2.73
1.6
Percent of total density represented by small
cladoceran individuals (SUBORDER=CLADOCERA
and CLADOCERAN_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD PDEN
samples combined)
1.03
3.34
-1.91
19.1
Biomass represented by native small cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCERAN_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD NAT BIO
samples combined)
2.985147667
31.79812541
-2.37
2.5
Density represented by native small cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCERA SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD NAT DEN
samples combined)
2.476364444
22.86662549
-1.99
2.2
Percent of total density represented by native small
cladoceran individuals (SUBORDER=CLADOCERA
and CLADOCERAN SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD NAT PDEN
samples combined)
1.03
3.33
-1.91
19.1
Density of individuals within the family Daphniidae
Cladoceran
DAPHNIID DEN
(coarse and fine net samples combined)
3.223097778
16.27482549
-2.09
2.5
Density of native individuals within the family
Cladoceran
DAPHNIID NAT DEN
Daphniidae (coarse and fine net samples combined)
3.223097778
16.27251961
-2.09
2.5
Density represented by individuals within the
subclass Copepoda (coarse and fine net samples
Copepod
COPE DEN
combined)
81.931315556
139.66798235
-1.74
1.5
Density represented by native individuals within the
subclass Copepoda (coarse and fine net samples
Copepod
COPE NAT DEN
combined)
81.931315556
139.66784314
-1.74
1.5
Number of distinct taxa within the copepod order
Copepod
CALAN NTAX
Calanoida (coarse and fine net samples combined)
1.3
1.1
2.10
2.4
Percent of total density represented by taxa of the
copepod order Calanoida (coarse and fine net
Copepod
CALAN PDEN
samples combined)
3.82
1.64
1.80
35.0
Number of distinct native taxa within the copepod
order Calanoida (coarse and fine net samples
Copepod
CALAN NAT NTAX
combined)
1.3
1.0
2.22
1.3
Percent of total density represented by individuals
of native taxa within the copepod order Calanoida
Copepod
CALAN NAT PDEN
(coarse and fine net samples combined)
3.81
1.64
1,80
35.0
Percent of distinct larger-sized native taxa
(NET_SIZECLS_NEW=COARSE; coarse and fine net
Richness/Diversity
COARSE NAT PTAX
samples combined)
40.65
37.17
1.64
0.3
212
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent total biomass from rotifers (coarse and fine
Rotifer
ROT PBIO
net samples combined)
23.72
34.91
-1.88
1.3
Percent of distinct taxa that are omnivorous (coarse
Trophic
OMNI PTAX
and fine net samples combined)
23.38
27.56
-2.36
1.6
Density of herbivorous cladocerans
(suborder=CLADOCERA; coarse and fine net
CLAD HERB DEN
samples combined)
6.8127244444
27.71694902
-1.94
1.9
Percent density represented by herbivorous
copepods (order=COPEPODA; coarse and fine net
COPE HERB PDEN
samples combined)
4.22
1.92
1.86
20.0
Metrics Derived from 300-count Subsamples of Coarse and Fine Net Samples
Percent of biomass represented by individuals of
taxa collected in coarse mesh net (150-um;
Abundance/
NET_SIZECLS_NEW=COARSE) in 300 count
Biomass/
subsamples (coarse and fine net samples
Density
COARSE300 PBIO
combined)
70.74
58.61
1.96
1.7
Percent of biomass represented by individuals of
native taxa collected in coarse mesh net (150-um;
Abundance/
NET_SIZECLS_NEW=COARSE) in 300 count
Biomass/
subsamples (coarse and fine net samples
Density
COARSE300 NAT PBIO
combined)
70.738666667
58.570196078
1.96
1.5
Percent biomass represented by individuals of
Abundance/
smaller-sized taxa (NET_SIZECLS_NEW=FINE) in
Biomass/
300-count subsamples (coarse and fine net samples
Density
FINE300 PBIO
combined)
29.26
41.39
-1.96
1.7
Biomass represented by large cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300 BIO
(coarse and fine net samples combined)
15.692285844
7.0078742941
2.02
1.4
Biomass represented by native large cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300 NAT BIO
(coarse and fine net samples combined)
15.692285844
7.0031208824
2.02
1.4
Biomass represented by small cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL) in 300-count subsamples
Cladoceran
SMCLAD300 BIO
(coarse and fine net samples combined)
1.8545441111
21.410646353
-2.40
2.6
Percent of small cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL) in 300-count subsamples
Cladoceran
SMCLAD300_PIND
(coarse and fine net samples combined)
10.90
19.03
-2.72
1.7
213
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of biomass represented by small
cladoceran individuals (SUBORDER=CLADOCERA
and CLADOCEAN_SIZE=SMALL) in 300-count
subsamples (coarse and fine net samples
Cladoceran
SMCLAD300 PBIO
combined)
5.50
16.12
-2.82
1.6
Biomass represented by native small cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL) in 300-count subsamples
Cladoceran
SMCLAD300 NAT BIO
(coarse and fine net samples combined)
1.8545441111
21.410646353
-2.40
2.5
Percent of native small cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL) in 300-count subsamples
Cladoceran
SMCLAD300 NAT PIND
(coarse and fine net samples combined)
10.90
19.03
-2.72
1.4
Number of distinct taxa within the copepod order
Calanoida in 300-count subsamples (coarse and fine
Copepod
CALAN300 NTAX
net samples combined)
1.3
1.0
1.94
2.8
Number of distinct native taxa within the copepod
order Calanoida in 300-count subsamples (coarse
Copepod
CALAN300 NAT NTAX
and fine net samples combined)
1.3
1.0
2.08
1.4
Percent distinct native taxa in 300-count subsample
Richness/Diversity
ZOCN300 NAT PTAX
of coarse net sample (150-um)
100
98.55
1.88
0.1
Number of distinct native taxa in coarse net
Richness/Diversity
ZOCN300 FAM NTAX
samples (150-um) based on 300-count subsample
5.1
4.7
1.47
0.8
Percent biomass from rotifers in 300-count
subsamples (coarse and fine net samples
Rotifer
ROT300 PBIO
combined)
22.26
34.91
-1.89
1.3
Percent of distinct taxa that are omnivorous in 300-
count subsamples (coarse and fine net samples
Trophic
OMNI300_PTAX
combined)
23.31
28.29
-2.60
1.5
214
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Table D. 3. List of candidate metrics used to develop the zooplankton MMI for the Plains bio-region
t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Abundance/
Percent of total biomass represented by individuals
Biomass/
collected in coarse mesh net (150-um for 2012
Density
COARSE PBIO
samples, 243 um for 2007 resamples)
57.38
70.00
-1.75
6.3
Abundance/
Percent of total biomass represented by native
Biomass/
individuals collected in coarse mesh net (150-um
Density
COARSE NAT PBIO
for 2012 samples, 243 um for 2007 resamples)
57.38
69.94
-1.74
6.3
Abundance/
Percent of biomass represented by individuals of
Biomass/
smaller-sized taxa (NET_SIZECLS_NEW=FINE; coarse
Density
FINE PBIO
and fine net samples combined)
42.62
30.00
1.75
6.3
Percent of biomass represented by native
Abundance/
individuals of smaller-sized taxa
Biomass/
(NET_SIZECLS_NEW=FINE; coarse and fine net
Density
FINE NAT PBIO
samples combined)
42.62
29.99
1.75
6.2
Percent of total individuals within the suborder
Cladocera that are "small"
(CLADOCERA_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD PIND
samples combined)
19.26
9.03
3.09
1.8
Percent of native individuals within the suborder
Cladocera that are "small"
(CLADOCERA_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD NAT PIND
samples combined)
19.26
8.94
3.11
1.8
Percent of total biomass represented by native
small cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL; coarse and fine net
Cladoceran
SMCLAD NAT PBIO
samples combined)
13.35
7.02
1.74
1.4
Copepod
Percent of total individuals within the subclass
COPE PIND
Copepoda (coarse and fine net samples combined)
29.45
41.97
-2.46
1.4
Copepod
Percent of native individuals within the subclass
COPE NAT PIND
Copepoda (coarse and fine net samples combined)
29.45
41.97
-2.46
1.4
Percent of distinct taxa that are within the copepod
order Calanoida (coarse and fine net samples
Copepod
CALAN PTAX
combined)
6.38
10.16
-2.32
2.0
Percent of total density represented by individuals
within the copepod order Calanoida (coarse and
Copepod
CALAN PDEN
fine net samples combined)
1.20
6.52
-2.06
14.1
Percent of total density represented by native
individuals within the copepod order Calanoida
Copepod
CALAN_NAT_PDEN
(coarse and fine net samples combined)
1.20
6.52
-2.06
14.1
215
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Ratio of Calanoid to (Cladoccera+Cyclopoids) based
on number of individuals (coarse and fine net
samples combined). Adapted from Kane et al.
(2009) Lake Erie plankton IBI. Calculated as
Copepod
COPE RATIO NIND
CALANOID_NIND/(CLAD_NIND+CYCLOPOID_NIND)
17.435
0.812
1.84
38.9
Ratio of Calanoid to (Cladoccera+Cyclopoids) based
on biomass (coarse and fine net samples
combined). Adapted from Kane et al. (2009) Lake
Erie plankton IBI. Calculated as
Copepod
COPE RATIO BIO
CALANOID BIO/(CLAD BIO+CYCLOPOID BIO)
7.325729723
1.327404241
2.31
4.6
Total distinct taxa richness (coarse and fine net
Richness/Diversity
TOTL NTAX
samples combined)
17.3
14..6
2.27
2.2
Total distinct native taxa richness (coarse and fine
Richness/Diversity
TOTL NAT NTAX
net samples combined)
17.3
14.5
2.34
2.2
Number of genera represented by distinct taxa
Richness/Diversity
GEN NTAX
(coarse and fine net samples combined)
13.8
11.6
2.45
2.2
Number of genera represented by distinct native
Richness/Diversity
GEN NAT NTAX
taxa (coarse and fine net samples combined)
13.8
11.5
2.56
2.2
Number of families represented by distinct taxa
Richness/Diversity
FAM NTAX
(coarse and fine net samples combined)
10.7
9.1
2.32
1.9
Number of families represented by distinct native
Richness/Diversity
FAM NAT NTAX
taxa (coarse and fine net samples combined)
10.7
9.1
2.41
2.2
Number of distinct taxa in fine net sample (ZOFN;
Richness/Diversity
ZOFN NTAX
80-um mesh)
12.4
9.8
2.69
1.7
Number of distinct native taxa in fine net sample
Richness/Diversity
ZOFN NAT NTAX
(ZOFN; 80-um mesh)
12. 4
9.8
2.73
1.7
Number of genera represented by distinct taxa in
Richness/Diversity
ZOFN GEN NTAX
fine net sample (ZOFN; 80-um mesh)
8.1
5.8
3.36
3.8
Number of genera represented by distinct native
Richness/Diversity
ZOFN GEN NAT NTAX
taxa in fine net sample (ZOFN; 80-um mesh)
8.1
5.8
3.42
3.8
Number of families represented by distinct taxa in
Richness/Diversity
ZOFN FAM NTAX
fine net sample (ZOFN; 80-um mesh)
6.6
4.7
3.48
3.0
Number of families represented by distinct native
Richness/Diversity
ZOFN FAM NAT NTAX
taxa in fine net sample (ZOFN; 80-um mesh)
6.6
4.7
3.56
3.0
Number of distinct taxa collected only in the fine-
Richness/Diversity
FINE NTAX
mish net (80-um; NET SIZECLS NEW=FINE)
10.5
8.0
2.61
1.8
Number of distinct native taxa collected only in the
Richness/Diversity
FINE NAT NTAX
fine-mish net (80-um; NET_SIZECLS_NEW=FINE)
10.5
8.0
2.63
1.7
Percent of total biomass represented in top 5 taxa
Richness/Diversity
DOM5 PBIO
(coarse and fine net samples combined)
91.31
94.16
-1.77
2.5
Number of distinct rotifer taxa (coarse and fine net
Rotifer
ROT_NTAX
samples combined)
10.5
8.0
2.63
1.7
216
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of total density represented by herbivorous
Trophic
COPE HERB PDEN
copepods (coarse and fine net samples combined)
1.23
6.58
-2.13
13.0
Metrics Derived from 300-count Subsamples of Coarse and Fine Net Samples
Percent of biomass represented by individuals of
Abundance/
taxa collected in coarse mesh net (150-um) in 300
Biomass/
count subsamples (coarse and fine net samples
Density
COARSE300 PBIO
combined)
59.0316
71.48616279
-1.77
5.2
Percent of biomass represented by native
Abundance/
individuals of taxa collected in coarse mesh net
Biomass/
(150-um) in 300 count subsamples (coarse and fine
Density
COARSE300 NAT PBIO
net samples combined)
59.0316
71.42267442
-1.76
5.1
Percent of biomass represented in individuals of
Abundance/
smaller-sized taxa (NET_SIZECLS_NEW=FINE) in the
Biomass/
300-count subsample (coarse and fine mesh
Density
FINE300 PBIO
samples combined)
42.15
28.64
1.89
6.0
Percent of biomass represented in native
individuals of smaller-sized taxa
Abundance/
(NET_SIZECLS_NEW=FINE) in the 300-count
Biomass/
subsample (coarse and fine mesh samples
Density
FINE300 NAT PBIO
combined)
42.15
28.63
1.90
5.8
Percent of small cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL) in 300-count subsamples
Cladoceran
SMCLAD300 PIND
(coarse and fine net samples combined)
19.788
9.848139535
2.97
2.0
Percent of biomass represented by small
cladoceran individuals (SUBORDER=CLADOCERA
and CLADOCEAN_SIZE=SMALL) in 300-count
subsamples (coarse and fine net samples
Cladoceran
SMCLAD300 PBIO
combined)
14.17
7.52
1.74
1.4
Percent of native small cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=SMALL) in 300-count subsamples
Cladoceran
SMCLAD300 NAT PIND
(coarse and fine net samples combined)
19.788
9.760930233
2.99
2.0
Percent of biomass represented by native small
cladoceran individuals (SUBORDER=CLADOCERA
and CLADOCEAN_SIZE=SMALL) in 300-count
subsamples (coarse and fine net samples
Cladoceran
SMCLAD300 NAT PBIO
combined)
14.17
7.47
1.76
1.4
Percent of individuals within the subclass Copepoda
in 300-count subsamples (coarse and fine net
Copepod
COPE300_PIND
samples combined)
30.94
43.16
2.42
1.3
217
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of native individuals within the subclass
Copepoda in 300-count subsamples (coarse and
Copepod
COPE300 NAT PIND
fine net samples combined)
30.94
43.16
30.93
1.3
Percent of distinct taxa within the copepod order
Calanoida in 300-count subsamples (coarse and fine
Copepod
CALAN300 PTAX
net samples combined)
7.51
11.20
-2.07
4.6
Ratio of Calanoid to (Cladoccera+Cyclopoids) based
on number of individuals in 300-count subsamples
(coarse and fine net samples combined). Adapted
from Kane et al. (2009) Lake Erie plankton IBI.
Calculated as
Copepod
COPE RATIO 300 NIND
CALANOID_NIND/(CLAD_NIND+CYCLOPOID_NIND)
12.675
0.800
1.83
19.6
Ratio of Calanoid to (Cladoccera+Cyclopoids) based
on biomass in 300-count subsamples (coarse and
fine net samples combined). Adapted from Kane et
al. (2009) Lake Erie plankton IBI. Calculated as
Copepod
COPE RATIO 300 BIO
CALANOID BIO/(CLAD BIO+CYCLOPOID BIO)
5.712
1.003
2.41
3.0
Total distinct native taxa richness in 300-count
subsamples (coarse and fine net samples
Richness/Diversity
TOTL300 NAT NTAX
combined)
14.8
12.9
1.76
1.4
Total distinct generic richness in 300-count
subsamples (coarse and fine net samples
Richness/Diversity
GEN300 NTAX
combined)
12.3
10.6
2.03
2.7
Total distinct native generic richness in 300-count
subsamples (coarse and fine net samples
Richness/Diversity
GEN300 NAT NTAX
combined)
12.3
10.5
2.13
2.9
Total distinct family richness in 300-count
subsamples (coarse and fine net samples
Richness/Diversity
FAM300 NTAX
combined)
9.8
8.4
2.11
2.3
Total distinct native family richness in 300-count
subsamples (coarse and fine net samples
Richness/Diversity
FAM300 NAT NTAX
combined)
9.8
8.4
2.22
2.6
Number of distinct genera in 300-count subsample
Richness/Diversity
ZOFN300 GEN NTAX
of fine-mesh net sample (50-^m)
6.8
5.3
2.45
2.7
Number of distinct native genera in 300-count
Richness/Diversity
ZOFN300 GEN NAT NTAX
subsample of fine-mesh net sample (50-^m)
6.8
5.2
2.48
2.9
Number of distinct families in 300-count subsample
Richness/Diversity
ZOFN300 FAM NTAX
of fine-mesh net sample (50-^m)
5.6
4.3
2.74
3.1
Number of distinct native families in 300-count
Richness/Diversity
ZOFN300 FAM NAT NTAX
subsample of fine-mesh net sample (50-^m)
5.6
4.3
2.79
3.1
Percent of biomass represented in top 5 taxa in
300-count subsamples (coarse and fine net samples
Richness/Diversity
DOM5 300 PBIO
combined)
91.38
94.27
-1.78
1.9
218
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Table D. 4. List of candidate metrics used to develop the zooplankton MMI for the Upper Midwest bio-region
t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Abundance/
Biomass/
Percent of native individuals (coarse and fine net
Density
TOTL NAT PIND
samples combined)
100
98.02
1.47
2348
Abundance/
Biomass/
Percent of density represented by native individuals
Density
ZOCN NAT PDEN
in coarse net sample (150-um)
100
95.90
1.52
Noise=0
Number of distinct taxa within the cladoceran
family Daphniidae (coarse and fine net samples
Cladoceran
DAPHNIID NTAX
combined)
1.4
1.8
-1.91
3.1
Density of individuals within the cladoceran family
Bosminidae (coarse and fine net samples
Cladoceran
BOSM DEN
combined)
28.20401905
6.857369231
1.85
2.8
Percent of individuals within the cladoceran family
Bosminidae (coarse and fine net samples
Cladoceran
BOSM PIND
combined)
15.31
8.35
1.85
19.5
Biomass of native individuals within the cladoceran
family Bosminidae (coarse and fine net samples
Cladoceran
BOSM NAT BIO
combined)
16.33606357
3.165346051
1.89
1.8
Density of native individuals within the cladoceran
family Bosminidae (coarse and fine net samples
Cladoceran
BOSM NAT DEN
combined)
28.204019048
5.0981051282
2.01
4.9
Percent of native individuals within the cladoceran
family Bosminidae (coarse and fine net samples
Cladoceran
BOSM NAT PIND
combined)
15.31
6.71
2.29
9.6
Percent of distinct native taxa within the
cladoceran family Bosminidae (coarse and fine net
Cladoceran
BOSM NAT PTAX
samples combined)
5.59
3.96
2.16
1.6
Percent of biomass represented by native
individuals within the cladoceran family Bosminidae
Cladoceran
BOSM NAT PBIO
(coarse and fine net samples combined)
10.01
2.57
2.07
4.9
Shannon Diversity based on the number of
cladoceran individuals (coarse and fine net samples
combined). Calculated as SUM{p(i)*Log[p(i)]},
where p(i) is proportion of individuals of taxon i,
Cladoceran
HPRIME CLAD
and Log= natural logarithm.
0.579
0.772
-1.91
1.3
Biomass of individuals within the copepod order
Copepod
CALAN BIO
Calanoida (coarse and fine net samples combined)
12.010544048
27.035772872
-1.73
12.7
Biomass of native individuals within the copepod
order Calanoida (coarse and fine net samples
Copepod
CALAN_NAT_BIO
combined)
12.010544048
27.025444897
-1.73
12.8
219
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of distinct native taxa (coarse and fine net
Richness/Diversity
TOTL NAT PTAX
samples combined)
100
98.05
2.65
21.7
Richness/Diversity
Percent of distinct taxa represented by native
ZOCN NAT PTAX
individuals in coarse net sample (150-um)
100
95.84
2.59
8.9
Richness/Diversity
Percent of distinct larger-sized taxa
(NET_SIZECLS_NEW=COARSE; coarse and fine net
COARSE PTAX
samples combined)
39.74
45.09
-1.89
1.4
Richness/Diversity
Percent of distinct smaller-sized taxa
(NET_SIZECLS_NEW=FINE; coarse and fine net
FINE PTAX
samples combined)
60.26
54.91
-1.89
1.4
Percent of distinct taxa within the phylum Rotifera
Rotifer
ROT PTAX
(coarse and fine net samples combined)
60.26
54.91
1.87
1.4
Density of individuals within the rotifer order
Flosculariaceae (coarse and fine net samples
Rotifer
FLOS DEN
combined)
290.0439619
115.22284872
1.82
7.6
Shannon Diversity based on the number of rotifer
individuals (coarse and fine net samples combined).
Calculated as SUM{p(i)*Log[p(i)]}, where p(i) is
proportion of individuals of taxon i, and Log=
Rotifer
HPRIME ROT
natural logarithm.
1.524
1.264
2.12
1.4
Simpson Diversity based on the number of rotifer
individuals (coarse and fine net samples combined).
Calculated as SUM{p(i)*p(i)} where p(i) is the
Rotifer
SIMPSON ROT
proportion of taxon 1 in the sample.
0.325
0.414
-1.79
2.4
Hurlbert's Probability of Interspecific Encounter
(PIE) based on the number of rotifer individuals
(coarse and fine net samples combined).
Calculated as SUM{p(i)*[N-n(i)/N-l]} where p(i) is
the proportion of taxon 1 in the sample, N is the
total number of rotifer individuals in the sample,
and n(i) is the number of rotifer individuals of taxon
Rotifer
PIE ROT
i in the sample.
0.678
0.590
1.76
2.5
Percent of rotifer individuals in top 3 Rotifer taxa
Rotifer
DOM3 ROT PIND
(coarse and fine net samples combined)
78.89
86.34
-2.35
1.6
Percent of rotifer individuals in top 5 Rotifer taxa
Rotifer
DOM5 ROT PIND
(coarse and fine net samples combined)
91.39
94.46
-1.81
2.6
Percent of rotifer biomass in dominant rotifer taxon
Rotifer
DOM1 ROT PBIO
(coarse and fine net samples combined)
45.30
59.27
-2.46
3.5
Percent of rotifer density in top 3 Rotifer taxa
Rotifer
DOM3 ROT PDEN
(coarse and fine net samples combined)
78.89
86.34
-2.35
1.6
Percent of density in top 5 rotifer taxa (coarse and
Rotifer
DOM5 ROT PDEN
fine net samples combined)
91.39
94.46
-1.81
2.6
220
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Metrics Derived from 300-count Subsamples of Coarse and Fine Net Samples
Number of distinct taxa within the cladoceran
family Daphniidae in 300-count subsamples (coarse
Cladoceran
DAPHNIID300 NTAX
and fine net samples combined)
1.2
1.7
-2.3
3.1
Number of distinct native taxa within the
cladoceran family Daphniidae in 300-count
subsamples (coarse and fine net samples
Cladoceran
DAPHNIID300 NAT NTAX
combined)
1.4
1.7
-2.3
3.1
Biomass of native individuals within the cladoceran
family Bosminidae in 300-count subsamples (coarse
Cladoceran
BOSM300 PIND
and fine net samples combined)
16.74
9.15
1.87
15.4
Density of native individuals within the cladoceran
family Bosminidae in 300-count subsamples (coarse
Cladoceran
BOSM300 NAT BIO
and fine net samples combined)
9.9940477143
2.211484641
1.84
2.1
Percent of native individuals within the cladoceran
family Bosminidae in 300-count subsamples (coarse
Cladoceran
BOSM300 NAT PIND
and fine net samples combined)
16.74
7.12
2.42
15.3
Percent of distinct native taxa that are within the
cladoceran family Bosminidae in 300-count
subsamples (coarse and fine net samples
Cladoceran
BOSM300 NAT PTAX
combined)
6.48
4.08
2.73
1.4
Biomass of biomass represented by native
individuals within the cladoceran family Bosminidae
in 300-count subsamples (coarse and fine net
Cladoceran
BOSM300 NAT PBIO
samples combined)
10.56
2.78
211
4.7
Biomass of individuals within the copepod order
Calanoida in 300-count subsamples (coarse and fine
Copepod
CALAN300 BIO
net samples combined)
6.3444415238
17.540568538
-2.17
9.2
Percent of distinct native taxa in 300-count
subsamples (coarse and fine net samples
Richness/Diversity
TOTL300 NAT PTAX
combined)
100
97.87
2.66
8.2
Percent of distinct native taxa in the coarse net
sample (150-um) based on the 300-individual
Richness/Diversity
ZOCN300 NAT PTAX
subsamples
100
95.92
2.76
Noise=0
Percent of distinct taxa represented by the rotifer
order Ploima in 300-count subsamples (coarse and
Rotifer
PLOIMA300 PTAX
fine net samples combined)
48.72
42.16
2.05
9.8
Shannon Diversity based on the number of rotifer
individuals in 300-count subsamples (coarse and
fine net samples combined). Calculated as
SUM{p(i)*Log[p(i)]}, where p(i) is proportion of
Rotifer
HPRIME ROT300
individuals of taxon i, and Log= natural logarithm.
1.515
1.254
2.12
1.4
221
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Simpson Diversity based on the number of rotifer
individuals in 300-count subsamples (coarse and
fine net samples combined). Calculated as
SUM{p(i)*p(i)} where p(i) is the proportion of taxon
Rotifer
SIMPSON ROT300
1 in the sample.
0.324
0.416
-1.86
2.1
Hurlbert's Probability of Interspecific Encounter
(PIE) based on the number of rotifer individuals in
300-count subsamples (coarse and fine net samples
combined). Calculated as SUM{p(i)*[N-n(i)/N-l]}
where p(i) is the proportion of rotifer taxon 1 in the
sample, N is the total number of rotifer individuals
in the sample, and n(i) is the number of individuals
Rotifer
PIE ROT300
of taxon i in the sample.
0.680
0.590
1,78
2.2
Percent of rotifer individuals in dominant rotifer
taxon in 300-count subsamples (coarse and fine net
Rotifer
D0M1 300 ROT PIND
samples combined)
45.70
54.61
-1.74
2.1
Percent of rotifer individuals in top 3 Rotifer taxa in
300-count subsamples (coarse and fine net samples
Rotifer
DOM3 300 ROT PIND
combined)
78.91
86.25
-2.26
1.4
Percent of rotifer individuals in top 5 Rotifer taxa in
300-count subsamples (coarse and fine net samples
Rotifer
DOM5 300 ROT PIND
combined)
91.50
94.71
-1.91
3.7
Percent of rotifer biomass in dominant Rotifer
taxon in 300-count subsamples (coarse and fine net
Rotifer
DOM1 300 ROT PBIO
samples combined)
47.97
58.94
-1.95
2.0
Percent of biomass represented by predator
individuals in 300-count subsamples (coarse and
Trophic
PRED300 PBIO
fine net samples combined)
2.06
0.93
1.86
95.5
Percent of biomass represented by predaceous
rotifer individuals in 300-count subsamples (coarse
Trophic
ROT PRED300 PBIO
and fine net samples combined)
2.06
0.93
1.86
95.5
Percent of biomass represented by herbivorous
Trophic
COPE_HERB_PBIO
copepods (coarse and fine net samples combined)
16.04
24.53
-1.96
5.0
222
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Table D. 5. List of candidate metrics used to develop the zooplankton MMI for the Western Mountains bio-region
t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of distinct native taxa within the
cladoceran family Bosminidae (coarse and fine net
Cladoceran
BOSM NAT PTAX
samples combined)
5.59
3.96
2.16
1.3
Number of distinct taxa within the subclass
Copepod
COPE NTAX
Copepoda (coarse and fine net samples combined)
2.6
3.3
-2.15
1.7
Percent of distinct taxa within the subclass
Copepod
COPE PTAX
Copepoda (coarse and fine net samples combined)
14.33
18.08
-2.29
1.9
Number of distinct native taxa within the subclass
Copepod
COPE NAT NTAX
Copepoda (coarse and fine net samples combined)
2.6
3.3
-2.07
1.7
Percent of distinct native taxa within the subclass
Copepod
COPE NAT PTAX
Copepoda (coarse and fine net samples combined)
14.33
18.00
-2.21
1.9
Total density of individuals within the subclass
Copepod
COPE DEN
Copepoda (coarse and fine net samples combined)
177.8479619
156.08843077
0.3
1.6
Total biomass of individuals within the copepod
order Calanoida (coarse and fine net samples
Copepod
CALAN BIO
combined)
12.010544048
27.035772872
-1.73
4.4
Total biomass of native individuals within the
copepod order Calanoida (coarse and fine net
Copepod
CALAN NAT BIO
samples combined)
12.010544048
27.025444897
-1.73
4.4
Percent of distinct larger-sized taxa
(NET_SIZECLS_NEW=COARSE; coarse and fine net
Richness/Diversity
COARSE PTAX
samples combined)
39.75
45.09
-1.87
2.3
Percent of distinct taxa collected only in the fine-
mesh net (50-um; NET_SIZECLS_NEW=FINE; coarse
Richness/Diversity
FINE PTAX
and fine net samples combined)
60.25
54.91
1.87
2.3
Simpson Diversity based on the total density
individuals (coarse and fine net samples combined).
Calculated as SUM{p(i)*p(i)} where p(i) is the
Richness/Diversity
SIMPSON DEN
proportion of density of taxon i in the sample.
0.288
0.353
-1.46
1.25
Percent distinct rotifer taxa (coarse and fine net
Rotifer
ROT PTAX
samples combined)
60.26
54.91
1.87
2.5
Percent distinct taxa that are within the rotifer
order Ploima (coarse and fine net samples
Rotifer
PLOIMA PTAX
combined)
48.72
42.00
2.28
4.3
Simpson Diversity based on the number of rotifer
individuals (coarse and fine net samples combined).
Calculated as SUM{p(i)*p(i)} where p(i) is the
Rotifer
SIMPSON ROT
proportion of taxon 1 in the sample.
0.325
0.414
-1.79
1.4
Percent of distinct taxa that are omnivorous
Trophic
COPE OMNI PTAX
copepods (coarse and fine net samples combined)
5.44
8.65
-2.526
1.5
223
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Metrics Derived from 300-count Subsamples of Coarse and Fine Net Samples
Abundance/
Total biomass of individuals in 300-count
Biomass/
subsamples (coarse and fine net samples
Density
TOTL300 BIO
combined)
90.072878905
270.55043706
-3.09
1.4
Abundance/
Total biomass of native individuals in 300-count
Biomass/
subsamples (coarse and fine net samples
Density
TOTL300 NAT BIO
combined)
90.072878905
269.19077886
-3.07
1.4
Abundance/
Biomass/
Biomass of individuals in 300-count subsample of
Density
ZOCN300 BIO
coarse net sample (150 um)
81.538501524
226.56640233
-2.68
2.2
Abundance/
Biomass/
Biomass of native individuals in 300-count
Density
ZOCN300 NAT BIO
subsample of coarse net sample (150 um)
81.538501524
225.20674414
-2.65
2.2
Biomass represented by individuals of large-sized
Abundance/
taxa in 300-count subsamples
Biomass/
(NET_SIZE_CLS=COARSE; coarse and fine net
Density
COARSE300 BIO
samples combined)
83.550340952
235.93896061
-2.77
3.0
Biomass represented by native individuals of large-
Abundance/
sized taxa in 300-count subsamples
Biomass/
(NET_SIZE_CLS=COARSE; coarse and fine net
Density
COARSE300 NAT BIO
samples combined)
62.150708119
234.5793024
-2.74
3.1
Percent biomass of native individuals of large-sized
Abundance/
taxa in 300-count subsamples
Biomass/
(NET_SIZE_CLS=COARSE; coarse and fine net
Density
COARSE300 NAT PBIO
samples combined)
85.15
75.20
1.88
5.7
Biomass of individuals within the suborder
Cladocera in 300-count subsamples (coarse and fine
Cladoceran
CLAD300 BIO
net samples combined)
62.150708119
173.03849657
-2.301
2.2
Biomass of native individuals within the suborder
Cladocera in 300-count subsamples (coarse and fine
Cladoceran
CLAD300 NAT BIO
net samples combined)
61.59444164
171.73934691
-2.28
2.2
Biomass represented by large cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300 BIO
(coarse and fine net samples combined)
54.826014262
142.47459983
-1.92
2.2
Percent of large cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300 PIND
(coarse and fine net samples combined)
20.42
14.14
2.22
1.8
Biomass represented by native large cladoceran
individuals (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300_NAT_BIO
(coarse and fine net samples combined)
54.826014262
142.37664379
-1.91
2.2
224
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t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignakNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of native large cladoceran individuals
(SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300 NAT PIND
(coarse and fine net samples combined)
20.41
13.47
2.49
1.8
Percent of distinct native taxa that are large
cladocerans (SUBORDER=CLADOCERA and
CLADOCEAN_SIZE=LARGE) in 300-count subsamples
Cladoceran
LGCLAD300 NAT PTAX
(coarse and fine net samples combined)
16.37
12.90
2.12
2.3
Biomass of individuals within the family Daphniidae
in 300-count subsamples (coarse and fine net
Cladoceran
DAPHNIID300 BIO
samples combined)
54.749187071
150.72825063
-2.08
3.0
Biomass of native individuals within the family
Daphniidae in 300-count subsamples (coarse and
Cladoceran
DAPHNIID300 NAT BIO
fine net samples combined)
54.749187071
150.63029459
-2.08
3.0
Total biomass of individuals within the subclass
Copepoda in 300-count subsamples (coarse and
Copepod
COPE300 BIO
fine net samples combined)
22.109055071
66.786813029
-2.76
2.0
Total biomass of native individuals within the
subclass Copepoda in 300-count subsamples
Copepod
COPE300 NAT BIO
(coarse and fine net samples combined)
22.109055071
66.726304529
-2.75
2.0
Total biomass of individuals within the copepod
order Calanoida in 300-count subsamples (coarse
Copepod
CALAN300 BIO
and fine net samples combined)
14.414470595
36.214300186
-2.00
3.2
Total biomass of native individuals within the
copepod order Calanoida in 300-count subsamples
Copepod
CALAN300 NAT BIO
(coarse and fine net samples combined)
14.414470595
36.153791686
-1.99
3.2
Number of distinct taxa in the 300-count subsample
Richness/Diversity
ZOFN300 NTAX
from the fine net sample (50-um)
7.3
8.4
-1.69
1.9
Simpson diversity based on number of individuals
Richness/Diversity
SIMPSON300 NIND
(coarse and fine net samples combined)
0.307
0.306
0.08
0
Percent of distinct taxa that are within the rotifer
family Asplanchnidae in 300-count subsamples
Rotifer
ASPLAN300 PTAX
(coarse and fine net samples combined)
0.88
2.25
-2.04
1.3
Biomass of herbivorous individuals in 300-count
subsamples (coarse and fine net samples
Trophic
HERB300 BIO
combined)
75.625607619
201.15711961
-2.56
3.1
Percent biomass of herbivorous individuals in 300-
count subsamples (coarse and fine net samples
Trophic
HERB300 PBIO
combined)
76.31
65.36
2.06
3.6
Number of distinct taxa that are omnivorous in 300-
count subsamples (coarse and fine net samples
Trophic
OMNI300 NTAX
combined)
3.0
3.6
-1.94
1.8
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NLA 2022 Technical Support Document - August 2024
t value
Mean Value for
Mean Value for
(Least disturbed vs.
Metric
Least disturbed
Most disturbed
Most disturbed
SignahNoise
Category
Metric Name
Description
Sites
Sites
Sites)
Value
Percent of distinct taxa that are predaceous
cladocerans in 300-count subsamples (coarse and
Trophic
CLAD PRED300 PTAX
fine net samples combined)
0.87
0
2.67
Noise=0
Percent biomass of herbivorous cladoceran
individuals in 300-count subsamples (coarse and
Trophic
CLAD HERB300 BIO
fine net samples combined)
62.140336143
173.03849657
-2.30
2.2
Biomass of omnivorous copepod individuals in 300-
count subsamples (coarse and fine net samples
Trophic
COPE OMNI300 BIO
combined)
4.7491737381
24.176607243
-2.38
2.0
Percent of distinct taxa represented by omnivorous
copepod individuals in 300-count subsamples
Trophic
COPE_OMNI300_PTAX
(coarse and fine net samples combined)
8.16
11.5
-2.15
2.1
226
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NLA 2022 Technical Support Document - August 2024
11.2 Non-target taxa in zooplankton samples that are excluded from enumeration
TAXA
ID
TAXON
NAME
PHYLUM
CLASS
SUBCLASS
ORDER
SUBORDER
FAMILY
GENUS
SPECIES
1026
AMPHIPODA
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
AMPHIPODA
1030
APPENDICULARIA
CHORDATA
APPENDICULARIA
1051
BIVALVIA
MOLLUSCA
BIVALVIA
1217
HEMIGRAPSUS
SANGUINEUS
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
DECAPODA
VARUNIDAE
HEMIGRAPSUS
SANGUINEUS
1359
MYSIDAE
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
MYSIDA
MYSIDAE
1389
PECTINARIIDAE
ANNELIDA
POLYCHAETA
PALPATA
CAN ALI PALPATA
TEREBELLIDA
PECTINARIIDAE
1390
PHYLLODOCIDAE
ANNELIDA
POLYCHAETA
PALPATA
ACICULATA
PHYLLODOCIDAE
1410
POLYCHAETA
ANNELIDA
POLYCHAETA
1461
TREMATODA
PLATYHELMINTHES
TREMATODA
1495
UCA
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
DECAPODA
PLEOCYEMATA
OCYPODOIDAE
UCA
5033
MYSIS RELICTA
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
MYSIDA
MYSIDAE
MYSIS
RELICTA
5049
GAMMARIDAE
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
AMPHIPODA
GAMMERIDEA
GAMMARIDAE
5491
CORBICULA
MOLLUSCA
BIVALVIA
HETERODONTA
VENEROIDA
CORBICULIDAE
CORBIUCLA
5497
DECAPODA
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
DECAPODA
5501
GAMMARUS
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
AMPHIPODA
GAMMERIDEA
GAMMARIDAE
GAMMARUS
5503
HYALELLA
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
AMPHIPODA
GAMMERIDEA
HYALELLIDAE
HYALELLA
HYALELLAGAMMARUS
5504
HYALELLA AZTECA
CMPLX
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
AMPHIPODA
GAMMERIDEA
HYALELLIDAE
HYALELLA
AZTECA CMPLX
5505
HYDRA
CNIDARIA
HYDROZOA
ANTHOATHECATAE
HYDRIDAE
HYDRA
5521
MONOCOROPHIUM
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
AMPHIPODA
GAMMERIDEA
COROPHIIDAE
MONOCOROPHIUM
5522
NEOMYSIS
MERCEDIS
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
MYSIDA
MYSIDAE
NEOMYSIS
MERCEDIS
5525
PALAEMONETES
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
DECAPODA
PLEOCYEMATA
PALAEMONIDAE
PALAEMONETES
5526
PALAEMONIDAE
ARTHROPODA
MALACOSTRACA
EUMALACOSTRACA
DECAPODA
PLEOCYEMATA
PALAEMONIDAE
5543
ANISOPTERA
ARTHROPODA
INSECTA
ODONATA
ANISOPTERA
227
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TAX A
ID
TAXON
NAME
PHYLUM
CLASS
SUBCLASS
ORDER
SUBORDER
FAMILY
GENUS
SPECIES
5544
CHAETONOTUS
GASTROTRICHA
CHAETONOTIDA
CHAETONOTIDAE
CHAETONOTUS
5545
COLLEMBOLA
ATRHROPODA
INSECTA
COLLEMBOLA
5546
CORIXIDAE
ARTHROPODA
INSECTA
PTERYGOTA
HEMIPTERA
HETEROPTERA
CORIXIDAE
5548
DIPTERA
ARTHROODA
INSECTA
DIPTERA
5549
DYTISCIDAE
ARTHROPODA
INSECTA
PTERYGOTA
COLEOPTERA
ADEPHAGA
DYTISICIDAE
5550
EPHEMEROPTERA
ARTHROPODA
INSECTA
EPHEMEROPTERA
5551
GASTROPODA
MOLLUSCA
GASTROPODA
5552
GASTROTRICHA
GASTROTRICHA
5554
INSECTA
ARTHROPODA
INSECTA
5555
NEMATODA
NEMATODA
5556
NEOGOSSEA
GASTROTRICHA
CHAETONOTIDA
NEOGOSSEIDAE
NEOGOSSEA
5557
NOTONECTIDAE
ARTHROPODA
INSECTA
PTERYGOTA
HEMIPTERA
HETEROPTERA
NOTONECTIDAE
5558
ODONATA
ARTHROPODA
INSECTA
ODONATA
5559
OLIGOCHAETA
ANNELIDA
CLITELLATA
OLIGOCHAETA
5562
PLECOPTERA
ARTHROPODA
INSECTA
PTERYGOTA
PLECOPTERA
5563
TARDIGRADA
TARDIGRADA
5564
TRICHOPTERA
ARTHROPODA
INSECTA
PTERYGOTA
TRICHOPTERA
5565
UNIONOIDA
MOLLUSCA
BIVALVIA
PALAEOHETERODONTA
UNIONOIDA
228
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