U.S. Department of the Interior

U.S. Geological Survey

Scientific Investigations Report 2008–5038

Guidelines for Design and Sampling for Cyanobacterial Toxin 

and Taste-and-Odor Studies in Lakes and Reservoirs
Photo 1

Photo 3

Photo 2

Front cover. Photograph 1: Beach sign warning of the presence of a cyanobacterial bloom, June 29, 2006 (photograph taken by 

Jennifer L. Graham, U.S. Geological Survey). Photograph 2: Sampling a near-shore accumulation of Microcystis, August 8, 2006 

(photograph taken by Jennifer L. Graham, U.S. Geological Survey). Photograph 3: Mixed bloom of Anabaena, Aphanizomenon, and 

Microcystis, August 10, 2006 (photograph taken by Jennifer L. Graham, U.S. Geological Survey). Background photograph: Near-shore 

accumulation of Microcystis, August 8, 2006 (photograph taken by Jennifer L. Graham, U.S. Geological Survey).
Guidelines for Design and Sampling for 

Cyanobacterial Toxin and Taste-and-Odor 

Studies in Lakes and Reservoirs

By Jennifer L. Graham, Keith A. Loftin, Andrew C. Ziegler, and Michael T. Meyer

Scientific Investigations Report 2008–5038

U.S. Department of the Interior

U.S. Geological Survey
U.S. Department of the Interior

DIRK KEMPTHORNE, Secretary

U.S. Geological Survey

Mark D. Myers, Director

U.S. Geological Survey, Reston, Virginia: 2008

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Suggested citation:

Graham, J.L., Loftin, K.A., Ziegler, A.C., and Meyer, M.T., 2008, Guidelines for design and sampling for cyanobacterial toxin and taste-and-odor studies in lakes and reservoirs: U.S. Geological Survey Scientific Investigations 

Report 2008–5038, 39 p.
iii

Contents

Abstract...........................................................................................................................................................1

Introduction.....................................................................................................................................................1

Purpose and Scope ..............................................................................................................................1

Background Information......................................................................................................................2

Light and Stratification in Lakes and Reservoirs....................................................................2

The Cyanobacteria ......................................................................................................................2

Cyanobacterial Toxins and Taste-and-Odor Compounds......................................................5

Taste-and-Odor Production by Actinomycetes Bacteria......................................................7

Detection of Cyanobacterial Toxins and Taste-and-Odor Compounds...............................8

Spatial and Temporal Variability of Cyanobacteria and Associated Toxins and Taste-and-Odor 

Compounds......................................................................................................................................10

Temporal Variability............................................................................................................................11

Spatial Variability in Lakes and Reservoirs ....................................................................................11

Spatial Variability in the Water Column...........................................................................................11

Common Study Types, Objectives, Designs, and Approaches for Studies of Cyanobacteria and 

Associated Toxins and Taste-and-Odor Compounds ...............................................................13

Reconnaissance Studies...................................................................................................................16

Monitoring Studies for Public Health and Drinking Water Treatment.......................................16

Interpretive Studies............................................................................................................................16

General Guidelines for Sampling ..............................................................................................................17

Surface Samples.................................................................................................................................17

Discrete-Depth Samples ...................................................................................................................18

Depth-Integrated Samples................................................................................................................18

Quality Control.....................................................................................................................................18

Sample Handling, Preparation, Processing, and Shipping for Analyses ..................................18

 Sample Handling...................................................................................................................18

 Processing Toxin and Taste-and-Odor Samples .............................................................18

 Processing Cyanobacterial (Phytoplankton) Samples ...................................................21

 Sample Shipping ...................................................................................................................21

Commonly Collected Ancillary Data .........................................................................................................22

Safety Considerations.................................................................................................................................22

U.S. Geological Survey National Water Information System Parameter Codes...............................22

Summary........................................................................................................................................................22

Acknowledgements.....................................................................................................................................24

Selected References...................................................................................................................................24

Glossary.........................................................................................................................................................29

Appendixes ...................................................................................................................................................31

1. Example Design and Approach for a Regional Reconnaissance Study............................32

2. Example Design and Approach for a Recreational Monitoring Study...............................35

3. Example Design and Approach for an Interpretive Study to Develop a Real-Time 

Model to Estimate Geosmin and 2-methylisoborneol (MIB) Concentrations ...................37
iv

Figures

1. Schematic showing stratification and light, temperature, and oxygen gradients that 

may develop in lakes and reservoirs.........................................................................................3

2. Photographs showing examples of cyanobacteria ................................................................4

3. Toxicity (lethal dose based on weight for 50 percent of the test population) of several 

cyanobacterial toxins...................................................................................................................7

4. Schematic showing the theoretical temporal distribution of total, particulate, and 

dissolved cyanobacterial toxin concentrations with respect to cyanobacterial 

population density ......................................................................................................................10

5. Photograph showing an example of spatial variability of cyanobacteria in a 

reservoir.......................................................................................................................................12

6. Schematic showing potential water column distributions of cyanobacteria...................13

7. Photographs showing the appearance of potential water column distributions of 

cyanobacteria .............................................................................................................................14

8. Schematic showing temperature and oxygen gradients that will develop in the water 

column when a metalimnetic cyanobacterial bloom is present.........................................16

9. Schematic showing the partitioning of cyanobacterial toxins and taste-and-odor 

compounds among the total, particulate, and dissolved phases in different types of 

samples ........................................................................................................................................22

Tables

1. Common genera of planktonic cyanobacteria that contain toxin and taste-and-odor 

producing strains..........................................................................................................................6

2. Common cyanobacterial toxins, toxicity (based on intraperitoneal mouse assays), and 

common effects of exposure ......................................................................................................8

3. Relative advantages and disadvantages of common analytical techniques utilized for 

analysis of cyanobacterial toxins and taste-and-odor compounds.....................................9

4. General study objectives and guidelines describing when, where, and how samples 

typically are collected for reconnaissance studies..............................................................19

5. General study objectives and guidelines describing when, where, and how samples 

typically are collected for monitoring studies .......................................................................20

6. General study objectives and guidelines describing when, where, and how samples 

typically are collected for interpretive studies assessing the physical, chemical, and 

biological factors affecting the occurrence and concentration of cyanobacteria and 

associated toxins and taste-and-odor compounds ..............................................................20

7. Samplers commonly used to collect surface, discrete depth, and depth integrated 

samples in lakes and reservoirs...............................................................................................21

8. Commonly collected ancillary data for studies of cyanobacterial toxins and taste-andodor compounds and references for additional information ..............................................23
v

SI to Inch/Pound

Multiply By To obtain

centimeter (cm) 0.3937 inch (in.)

cubic micrometer (μm3) 6.102 x 10-14 cubic inch (in3)

cubic micrometers per milliliter (μm3/mL) 1.805 x 10-12 cubic inch per ounce

liter (L) 33.82 ounce, fluid (fl. oz)

liter (L) 2.113 pint (pt)

liter (L) 1.057 quart (qt)

liter (L) 0.2642 gallon (gal)

meter (m) 3.281 foot (ft) 

meter (m) 1.094 yard (yd) 

microgram per gram (μg/g) 1.600 x 10-6 ounce per pound (oz/lb)

microgram per kilogram (μg/kg) 1.600 x 10-8 ounce per pound (oz/lb)

microgram per liter (μg/L) 1.0 part per billion (ppb)

micrometer (μm) 3.937 x 10-6 inch (in.)

milliliter (mL) 0.0338 ounce, fluid (oz)

nanogram per liter (ng/L) 1.0 part per trillion (ppt)

Temperature in degrees Fahrenheit (°F) may be converted to degrees Celsius (°C) as follows:

°C=(°F-32)/1.8

Temperature in degrees Celsius (°C) may be converted to degrees Fahrenheit (°F) as follows:

°F=(1.8×°C)+32.

Abstract

Cyanobacteria and associated toxins and taste-and-odor 

compounds are of increasing environmental concern. However, consistent guidelines for the development of studies 

assessing cyanobacterial toxins and taste-and-odor compounds 

presently are not available. This report provides guidance for 

the development of scientific studies of cyanobacteria and 

associated by-products in lakes and reservoirs. Topics include: 

background information on cyanobacteria, toxins, and tasteand-odor compounds; spatial and temporal considerations 

that are unique to the cyanobacteria in lakes and reservoirs; 

common study types, objectives, and approaches for studies 

of cyanobacteria and associated toxins and taste-and-odor 

compounds; general guidelines for collecting samples; and 

information on sample handling, preparation, processing, 

and shipping.

Introduction

Cyanobacteria (also referred to as blue-green algae) cause 

a multitude of water-quality concerns, including the potential 

to produce toxins and taste-and-odor compounds. Toxins and 

taste-and-odor compounds may cause significant economic 

and public health concerns, and are of particular interest in 

lakes, reservoirs, and rivers that are used for drinking-water 

supply, recreation, or aquaculture. Cyanobacterial toxins 

(cyanotoxins) have been implicated in human and animal illness and death in more than 50 countries worldwide, including at least 27 states in the United States (Yoo and others, 

1995; Chorus and Bartram, 1999; Huisman and others, 2005). 

Human toxicoses associated with cyanobacterial toxins most 

commonly have occurred after exposure through drinking 

water or recreational activities (Yoo and others, 1995; Chorus and Bartram, 1999; Falconer, 2005; Huisman and others, 2005; Graham, 2006). Taste-and-odor compounds cause 

malodorous or unpalatable drinking water and fish flesh, 

resulting in increased treatment costs and loss of recreational 

or aquacultural revenue. Federal and state agencies, resource 

managers, drinking-water treatment-facility operators, lake 

associations, and local officials increasingly are faced with 

decisions about managing cyanobacterial blooms that affect 

local economies and public awareness, exposure, and health; 

therefore, representative scientific data are needed to guide 

management and public health decisions about cyanobacterial 

toxins and taste-and-odor compounds.

The study of cyanobacterial toxins and taste-and-odor 

compounds is an active area of developing research. Many 

different approaches have been used in the design, sample collection, and analysis of studies addressing these cyanobacterial 

by-products. Consistent guidelines for the development of 

such studies presently (2008) are not available, often making 

cross-comparisons difficult (Graham and Jones, 2007).

As part of its mission, the U.S. Geological Survey 

(USGS) collects the data needed to assess the quality of our 

Nation’s water resources. A key aspect of this mission is 

consistent use of field methods for the purpose of producing nationally comparable data. There is a need for citable 

documentation for USGS water-quality data-collection 

protocols for scientific studies of cyanobacteria and associated 

by-products in lakes and reservoirs.

Purpose and Scope

The purpose of this report is to provide guidelines for 

design and sampling for cyanobacterial toxin and taste-andodor compounds in lakes and reservoirs. This report: provides 

general background information on cyanobacteria and associated toxins and taste-and-odor compounds; discusses special 

considerations for study design and sampling approaches that 

are unique to planktonic cyanobacteria and associated toxin 

and taste-and-odor compounds; provides guidance on appropriate sampling approaches for the system being studied; and 

provides guidance on sample collection, handling, preparation, 

processing, and shipping for laboratory analysis of toxins and 

taste-and-odor compounds, and algal identification, enumeration, and biovolume.

The fundamentals of study design are beyond the scope 

of this report, but there are special characteristics of planktonic 

cyanobacteria that need to be considered when developing 

cyanobacterial toxin and taste-and-odor studies. The information provided in this report is intended to be used in conjunction with other reports that specifically address the design of 

Guidelines for Design and Sampling for Cyanobacterial 

Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

By Jennifer L. Graham, Keith A. Loftin, Andrew C. Ziegler, and Michael T. Meyer 
2 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

surface-water-quality studies (Friedman and Erdmann, 1982; 

Shampine and others, 1992; Averett and Schroder, 1994).

In addition to planktonic cyanobacteria, benthic 

cyanobacteria in lakes, reservoirs, streams, and rivers also may 

produce toxins and taste-and-odor compounds (Chorus and 

Bartram, 1999; Falconer, 2005; Huisman and others, 2005; 

Taylor and others, 2005). Although consistent guidelines 

presently (2008) are not available for collecting samples and 

designing studies of benthic cyanobacteria, sampling this component in aquatic ecosystems, especially streams, is beyond 

the scope of this report. Other algal groups besides cyanobacteria also may cause taste-and-odor problems in drinkingwater supplies; however, the most frequent and severe events 

are associated with cyanobacteria (Wnorowski, 1992; Rashash 

and others, 1996; Taylor and others, 2005). One other freshwater/brackishwater alga (the haptophyte Prymnesium parvum) 

is known to produce icthyotoxins, but these toxins have been 

associated with fish kills only, and not human illness (Sager 

and others, 2007). All approaches presented in this report are 

discussed with an emphasis on the cyanobacteria; but these 

procedures also may be applicable for studies of other planktonic algal groups, depending on study objectives.

Background Information

Light and Stratification in Lakes and Reservoirs

Lakes and reservoirs are characterized by vertical gradients caused by light and thermal stratification. An understanding of these gradients is required when designing studies and 

sampling strategies for lakes and reservoirs. Key terms and 

concepts are described herein. For more detail on light and 

thermal stratification in lakes and reservoirs see: Horne and 

Goldman (1994), Lampert and Sommer (1997), Wetzel and 

Likens (2000), and Wetzel (2001).

Light intensity decreases exponentially with depth, and a 

light gradient is present in all lakes (fig. 1) (Horne and Goldman, 1994; Lampert and Sommer, 1997; Wetzel and Likens, 

2000; Wetzel, 2001). Common terms used to describe light 

gradients in lakes include:

Photic (Euphotic) Zone—Region where there is enough 

light to support photosynthesis; extends from the surface to 

the depth where light is approximately 1 percent of that at the 

surface.

Aphotic Zone—Region where there is not enough light to 

support photosynthesis; extends from below the photic zone to 

the lake bottom.

Thermal stratification creates isolated layers that result in 

thermal and chemical (such as nutrient) gradients; stratification is the result of differences in density associated with temperature (fig. 1). Stratification in deep lakes tends to be stable, 

whereas daily or continual mixing may occur in shallow lakes. 

Thermal stratification is a summer phenomenon in temperate 

lakes, but may occur year round in sub-tropical and tropical lakes. Spatial variability of stratification within lakes and 

reservoirs is common because of changes in depth and other 

morphological features (Horne and Goldman, 1994; Lampert 

and Sommer, 1997; Wetzel and Likens, 2000; Wetzel, 2001). 

Common terms used to describe stratification include:

Epilimnion—The warm, buoyant upper layer of a stratified lake.

Metalimnion—The middle layer of a stratified lake; the 

metalimnion is characterized by substantial decreases in temperature with depth.

Thermocline—The region where temperature change is 

greater than or equal to 1°C per meter; the terms thermocline 

and metalimnion often are used synonymously.

Hypolimnion—The cold, dense bottom layer of a stratified lake; the hypolimnion often becomes anoxic (little or no 

dissolved oxygen) in productive systems.

Turnover—Complete isothermal mixing of a previously 

stratified lake. 

Mixed Depth—The depth of turbulent mixing; may 

include all or only a part of the water column depending on 

stratification, solar irradiance, and wind.

During calm periods temporary mixed layers (lasting 

hours-weeks) may form in shallow lakes, deeper lakes that do 

not typically stratify, or the epilimnion of stratified lakes, creating greater complexity in vertical structure. Mixing in shallow and stratified lakes will vary daily and seasonally depending on solar irradiance and wind. Thermal stratification is most 

common, but gradients in water chemistry (often caused by 

submerged springs), such as salinity, also may cause stratification (Horne and Goldman, 1994; Lampert and Sommer, 1997; 

Wetzel and Likens, 2000; Wetzel, 2001). 

The Cyanobacteria

Cyanobacteria (fig. 2) are true bacteria with a prokaryotic 

cell structure; however, cyanobacteria also have chlorophyll-a, 

a photopigment characteristic of eukaryotic algae and higher 

plants. Structurally the cyanobacteria are bacteria-like, but 

functionally the cyanobacteria are algae-like. Because of 

this photosynthetic functionality, cyanobacteria typically are 

sampled and analyzed as part of phytoplankton (algal) assemblages rather than bacterial assemblages in aquatic ecosystems 

(Wetzel, 2001).

Cyanobacteria are a natural part of phytoplankton assemblages in lakes and reservoirs and commonly are present in 

at least low abundances (Reynolds, 1984; Jones and Korth, 

1995). Cyanobacterial abundance and community composition 

vary seasonally as a result of changes in water temperature, 

solar irradiance, meteorological conditions, hydrology, and 

nutrient supply. In temperate climates, cyanobacteria typically 

dominate the phytoplankton during mid-summer to early fall, 

but may become dominant any time throughout the year, even 

under ice during winter. In subtropical and tropical climates 

cyanobacteria may dominate at any time, and dominance may 

persist year-round (Chorus and Bartram, 1999; Wetzel, 2001; 

Falconer, 2005; Huisman and others, 2005).
Introduction 3

Eutrophication results in conditions that are favorable for 

cyanobacterial growth (elevated nutrients, reduced light penetration) and cyanobacterial blooms frequently occur in eutrophic (nutrient rich, high productivity) lakes and reservoirs. 

Cyanobacterial blooms also may occur in oligotrophic (nutrient poor, low productivity) systems, although not as frequently 

as in eutrophic systems. Blooms in oligotrophic systems often 

are associated with benthic mats of cyanobacteria or favorable nutrient and light conditions at depth in a stably stratified 

water column (Reynolds, 1984; Chorus and Bartram, 1999; 

Wetzel, 2001; Falconer, 2005; Huisman and others, 2005).

Many, but not all, cyanobacteria have gas vacuoles that 

allow them to maintain a favorable position in the water column by regulating buoyancy. Light (photic depth), nutrients, 

carbon availability, stratification, and mixed depth (depth of 

turbulent mixing in the water column) all affect cyanobacterial 

position in the water column. Optimal water-column position 

is species specific. There are three distinct patterns in distribution that may develop (Reynolds and Walsby, 1975; Reynolds, 

1987; Humphries and Lyne, 1998):

1. Cyanobacteria may maintain a position in the photic 

zone, regardless of mixed depth. Populations typically 

are uniformly distributed throughout the photic zone. 

This distribution may develop in well-mixed lakes or the 

epilimnion of stably stratified lakes. Many cyanobacterial genera, including Anabaena, Aphanizomenon, 

Cylindrospermopsis, and Microcystis display this type of 

buoyancy regulation. 

2. Cyanobacteria may migrate to different locations in the 

photic zone throughout the day. Populations generally 

move towards the surface at night or early morning and 

downwards later in the day. Diel movement of cyanobacterial populations typically is linked with the development 

of diel mixed layers in shallow lakes or the epilimnion of 

stably stratified lakes. Regardless of movement or location, cyanobacteria typically maintain a position in the 

photic zone. In shallow lakes, where light penetrates to 

the bottom, cyanobacteria may spend part of the day on 

the sediment surface. Diel movement most commonly is 

associated with species of Microcystis. 

Increasing depth

EPILIMNION

HYPOLIMNION

Not to scale

METALIMNION (THERMOCLINE)

Light

Surface

Bottom

PHOTIC DEPTH

PHOTIC

ZONE

APHOTIC

ZONE

Temperature

Oxygen

Increasing values

Increasing values

Figure 1. Stratification and light, temperature, and oxygen gradients that may develop in lakes and reservoirs.
4 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Unicellular Cyanocatena

(Photo courtesy of A. St. Amand, Phycotech, Inc.)

Colonial Microcystis

(Photo courtesy of A. St. Amand, Phycotech, Inc.)

Filamentous Anabaena and Aphanizomenon

(Photo courtesy of A. St. Amand, Phycotech, Inc.)

B.

A.

C.

D.

Anabaena

Aphanizomenon

Visible colonies of Microcystis near the

water surface

(Photo by J. Graham, USGS)

Picoplankton

Single Cells

10μm

10μm

10μm

Figure 2. Examples of cyanobacteria: Cyanobacteria may be unicellular (A), colonial (B), or filamentous (C) and range in size from 

0.2 micrometers to greater than 1,600 micrometers. Larger filaments and colonies are often visible to the naked eye (D).
Introduction 5

3. Cyanobacteria may maintain a position at a particular 

depth, typically at the interface between the epilimnion 

and metalimnion. This phenomenon is referred to as a 

metalimnetic bloom, and most commonly is associated 

with species of Planktothrix (Oscillatoria). Metalimnetic cyanobacterial blooms occur most frequently in 

mesotrophic (moderate nutrients and productivity) lakes. 

For metalimnetic blooms to develop, the water column 

must be stably stratified and photic depth must penetrate 

into the metalimnion; metalimnetic cyanobacterial blooms 

can exist at light levels at or below one percent of incident 

light. 

The term bloom, not specific to cyanobacteria, is inexact 

and subjective (Smayda, 1997). The simplest definitions are: 

a large population or extremely high cell densities of phytoplankton (extremely high densities are typically defined as 

greater than 20,000 to 100,000 cells per milliliter); where proliferation of phytoplankton is dominated by a single or a few 

species; or visible accumulation of phytoplankton at the water 

surface (Cottingham and others, 1995; Chorus and Bartram, 

1999; Dodds, 2002; Huisman and others, 2005; Falconer and 

Humpage, 2006; Jacoby and Kann, 2007).

When referring to cyanobacteria, the term bloom commonly is associated with the accumulation of cyanobacteria at 

the water surface; however, cyanobacterial blooms are more 

typically mixed throughout the photic zone, epilimnion, or 

water column. Surface accumulations, or scums, may develop 

when cyanobacteria float to the surface during unusually calm 

conditions that cause a sudden change in turbulent mixing 

(sudden calm will not affect metalimnetic populations) or 

when cyanobacteria begin to senesce (age) and are no longer 

able to effectively regulate buoyancy. Surface accumulations 

may develop and dissipate within a matter of hours as conditions change. Because the entire cyanobacterial population 

may accumulate within 1 to 2 centimeters of the water surface, 

cell densities may be extremely large (more than a million 

cells per milliliter), even if water-column densities were 

relatively small (less than 20,000 cells per milliliter). Surface 

accumulations of cyanobacteria may concentrate even further 

when wind blows to leeward areas (for example, near-shore 

areas, bays, and inlets), often resulting in the development 

of dense accumulations that may extend from the surface to 

depths of more than 1 meter (Reynolds and Walsby, 1975; 

Reynolds, 1987; Soranno, 1997; Humphries and Lyne, 1998; 

Chorus and Bartram, 1999; Dodds, 2002; Falconer, 2005; 

Huisman and others, 2005; Jacoby and Kann, 2007). 

Cyanobacterial Toxins and Taste-and-Odor 

Compounds

Cyanobacterial toxins and taste-and-odor compounds are 

naturally produced algal by-products; however, the function 

of these compounds currently (2008) is unknown. Hypothesized functions include primary roles in cellular processes, 

secondary metabolites, allelopathy, or defense mechanisms. 

Production of toxins and taste-and-odor compounds is strain, 

rather than species, dependent. Any one cyanobacterial 

species may have multiple strains, and toxic and non-toxic 

strains may occur simultaneously in an individual lake (Vezie 

and others, 1998). Most cyanobacterial taxa do not produce 

toxins or taste-and-odor compounds, but many of the common 

planktonic genera contain one or more toxin and/or taste-andodor producing strains (table 1). Whereas some strains may 

produce toxin and taste-and-odor compounds simultaneously, 

these compounds do not necessarily co-occur and the presence and concentration of one may not be reliably used to 

predict the presence and concentration of another (Chorus and 

Bartram, 1999). Because toxin and taste-and-odor production 

is strain dependent, algal identification alone cannot be used 

to determine whether or not these by-products will be present, 

although genera that contain strains producing these compounds can be identified. Likewise, co-occurrence of potential 

producers and toxin and/or taste-and-odor compounds does 

not positively identify the actual producer. Strain isolation and 

culture are required to conclusively determine the producer(s) 

of measured toxin and/or taste-and-odor compounds (Chorus 

and Bartram, 1999; Watson, 2003; Falconer, 2005; Huisman 

and others, 2005; Taylor and others, 2005). Genetic techniques that isolate specific gene clusters also are promising 

in identifying toxin producers in complex environmental 

samples (Nonneman and Zimba, 2002; Hisbergues and others, 2003; Kurmayer and Kutzenberger, 2003; Vaitomaa and 

others, 2003).

Cyanobacterial toxins are chemically and bioactively 

diverse (Sivonen, 1996; Carmichael, 1997; Chorus and Bartram, 1999), all targeting fundamental cellular processes, and 

thereby affecting a wide range of organisms (Falconer, 1993; 

Christofferson, 1996). The cyanobacteria are the only member 

of freshwater phytoplankton communities known to produce 

toxins that also may affect terrestrial organisms. The three 

main classes of cyanobacterial toxins, defined by their effects 

on human health, are dermatoxins, neurotoxins, and hepatotoxins (Chorus, 1993; Chorus and Bartram, 1999; Falconer, 

1999). Dermatoxins cause acute, often severe, dermatitis after 

contact with cyanobacteria, and also may cause inflammation 

of the gastro-intestinal tract (Volterra, 1993; Chorus, 1993; 

Sivonen, 1996; Chorus and Bartram, 1999; Stewart and others, 

2006). Neurotoxins, which affect the central nervous system, 

and hepatotoxins, which affect the liver, are extremely toxic, 

and numerous wild and domestic animal kills have been linked 

to such blooms. Whereas neurotoxins rarely are associated 

with human illness and death, human toxicoses associated 

with hepatotoxins have occurred after consumption of drinking water, contact with cyanobacteria during recreational 

activities (including inhalation exposure), and haemodialysis 

using tainted water (Chorus, 1993; Volterra, 1993; Carmichael 

and others, 2001; Chorus and Bartram, 1999; Falconer, 2005; 

Huisman and others, 2005). 

The most common cyanobacterial toxin groups are 

described in figure 3 and table 2. Many of the cyanobacterial 

toxins have multiple variants, with some variants being more 
6 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

toxic than others. For example, the microcystins, currently 

believed to be the most common group of cyanobacterial toxins, have more than 80 known variants with LD50’s (lethal dose 

required to kill 50 percent of the test population; determined 

by intraperitoneal mouse assays) ranging from 25 to greater 

than 1,000 micrograms per kilogram (μg/kg) (most variants 

fall within the 50–300 μg/kg range). The adverse health effects 

caused by contact with, or ingestion or inhalation of, large 

concentrations of cyanobacterial toxins (acute exposure) are 

well documented (table 2). The effects of exposure to low 

levels of cyanobacterial toxins for an extended period of time 

(chronic exposure) are not well understood. Several of the 

dermatoxins and hepatotoxins also are known to be potent 

tumor promoters (Chorus and Bartram, 1999); however, the 

effects of chronic exposure to low doses of cyanobacterial 

toxins with time are largely unknown. The recently discovered 

neurotoxin β-N-methylamino-L-alanine (BMAA) is unique 

in that it does not have known acute effects on human health, 

but chronic exposure to BMAA may be linked to neurodegenerative diseases such as ALS (amyotrophic lateral sclerosis), 

Parkinson’s, and Alzheimer’s (Cox and others, 2005). Unlike 

toxins, cyanobacterial taste-and-odor compounds have no 

known effects on human health (Watson, 2003).

The greatest risk of exposure to elevated concentrations 

of cyanobacterial toxins probably is through accidental ingestion and inhalation during recreational activities. The World 

Health Organization (WHO) has defined low, moderate, high, 

and very high risk categories for adverse health effects that 

occur through recreational exposure to microcystins. Based on 

the WHO risk categories for recreational exposure, microcystin concentrations less than 10 micrograms per liter (μg/L) 

have a low risk, 10–20 μg/L a moderate risk, 20–2000 μg/L 

a high risk, and greater than 2000 μg/L a very high risk for 

adverse health effects (Chorus and Bartram, 1999). WHO recreational guidelines do not distinguish between children and 

adults, although children are considered to be at greater risk 

because of low body mass and their tendency to ingest water 

while playing (Chorus, 2001). 

The WHO also has developed a provisional guideline of 

1 μg/L for microcystin-LR in finished drinking water (Chorus and Bartram, 1999) and currently (2008) is developing a 

guideline for cylindrospermopsin. Falconer (2005) has proposed a guideline of 1 μg/L for cylindrospermopsin in finished 

drinking water. Several countries have set national standards 

or guidelines for microcystin in drinking water based on 

the WHO guideline, including Australia, Brazil, Canada, 

the Czech Republic, France, Poland, and Spain (Huisman and 

Table 1. Common genera of planktonic cyanobacteria that contain toxin and taste-and-odor producing strains. 

[All data included in this table are based on documented production in laboratory cultures; data based on circumstantial evidence, such as co-occurrence of 

genera and toxin or taste-and-odor compounds in environmental samples, were not included in this table. LYN, lyngbyatoxin-a; APL, aplysiatoxins; LPS, 

lipopolysaccharides; CYL, cylindrospermopsins; MC, microcystins; NOD, nodularins; ATX, anatoxins; BMAA, β-N-methylamino-L-alanine; NEO, neosaxitoxins; SAX, saxitoxins; GEOS, geosmin; MIB, 2-methylisoborneol]

Cyanobacterial Genera

Dermatoxins Hepatotoxins Neurotoxins Tastes and 

odors

LYN APL LPS CYL MC NOD ATX BMAA NEO SAX GEOS MIB

Colonial/filamentous

 Anabaena X X X X X X X X

 Anabaenopsis X X

 Aphanizomenon X X X X X X X X

 Aphanocapsa X X

 Cylindrospermopsis X X X X

 Microcystis X X X

 Nodularia X X X

 Oscillatoria (Planktothrix) X X X X X X X X X

 Pseudanabaena X X X X

 Raphidiopsis X X X

Unicellular

 Synechococcus X X X X X

 Synechocystis X X X

Sources: Wu and others (1991), Wnorowski (1992), Blevins and others (1995), Carmichael (1997), Bláha and Maršálek (1999), Chorus and Bartram (1999), 

Domingos and others (1999), Saadoun and others (2001), Oudra and others (2002), Watson (2003), Huisman and others (2005), and Taylor and others (2005). 

A comprehensive list of known cyanobacterial toxin and taste-and-odor producers is not currently (2008) available in the literature. Combined, the references 

used to create this table may be used to create a fairly complete list of planktonic and benthic producers.
Introduction 7

others, 2005), and in the United States the cyanobacterial toxins microcystin, cylindrospermopsin, and anatoxin currently 

are on the U.S. Environmental Protection Agency (2005) 

drinking-water contaminant candidate list. 

There are many potential sources of taste-and-odor in 

finished drinking water, including biological activity in the 

source water, chemical contamination (natural and anthropogenic) of the source water, chemicals used in treatment 

processes, and biological activity or materials present in 

the distribution system. Biological activity associated with 

naturally occurring algae in source water is among the most 

common causes of tastes and odors in finished drinking water 

(Rashash and others, 1996; Taylor and others, 2005; Watson, 

2003; Wnorowski, 1992). Many groups of algae produce tasteand-odor compounds, with approximately 200 different compounds identified to date (2008) (Rashash and others, 1996; 

Watson, 2003; American Water Works Association, 2004); 

however, most taste-and-odor problems in drinking water are 

associated with cyanobacterial production of geosmin and 

2-methylisoborneol (MIB) (Wnorowski, 1992; Rashash and 

others, 1996; Taylor and others, 2005). 

The taste-and-odor compounds geosmin and MIB cause 

earthy/musty tastes and odors and are detectable by humans at 

concentrations between 5–10 nanograms per liter; therefore, 

these compounds may be detectable in the environment before 

potential cyanobacterial producers are detected. In addition, 

the producer may be a relatively small component of the 

phytoplankton community. Taste-and-odor episodes caused by 

cyanobacteria have occurred even when cyanobacteria are not 

at detectable levels in the water column (Wnorowski, 1992; 

Jones and Korth, 1995; Rashash and others, 1996; Watson, 

2003; Taylor and others, 2005). 

Taste-and-Odor Production by Actinomycetes 

Bacteria

Most taste-and-odor problems associated with geosmin 

and/or MIB are caused by cyanobacteria but bacteria in the 

actinomycetes group also may produce geosmin and MIB 

(Wnorowski, 1992; Rashash and others, 1996; Watson, 2003; 

Taylor and others, 2005). Actinomycetes bacteria are not 

photosynthetic and are not part of the phytoplankton community in lakes and reservoirs. The actinomycetes bacteria 

largely are terrestrial organisms associated with soils; tasteand-odor compounds typically are produced terrestrially and 

washed into lakes or reservoirs with or without the bacteria 

that produced them. Inflow events with high suspendedsediment loads may result in taste-and-odor episodes caused 

by geosmin/MIB produced by actinomycetes bacteria (Zaitlin 

and others, 2003; Zaitlin and Watson, 2006). When washed 

in, actinomycetes bacteria generally are associated with the 

sediment. However, taste-and-odor compounds produced by 

actinomycetes bacteria can be located in the water column 

and throughout the lake or reservoir, depending on whether 

the compounds are dissolved or associated with the bacteria, 

hydrology of inflows and mixing with surrounding water, and 

sediment settling rates. Because of settling, actinomycetes may 

accumulate in the hypolimnion or near the bottom of lakes or 

reservoirs. It currently (2008) is unknown if or for how long 

terrestrial actinomycetes bacteria remain metabolically active 

Algal Toxins

SAXITOXIN

ANATOXIN-A(S)

ANATOXIN-A

MICROCYSTINS -LR, -LA, -YR

MICROCYSTIN-RR

CYLINDROSPERMOPSIN (24 H)

CYLINDROSPERMOPSIN (5 D)

10,000

1,000

100

10

1

Lethal dose required to kill half the members of a tested

population, in micrograms per kilogram body weight

(Microcystins -LR, -LA, and -YR indicates three

microcystin variants of similar toxicity)

Figure 3. Toxicity (lethal dose based on weight 

for 50 percent of the test population) of several 

cyanobacterial toxins. Toxicity is based on 

intraperitoneal mouse assays (table 2). Figure 

based on data from Chorus and Bartram (1999).
8 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

in aquatic environments (Wnorowski, 1992; Rashash and others, 1996; Watson, 2003; Zaitlin and others, 2003; Taylor and 

others, 2005; Zaitlin and Watson, 2006). 

The actinomycetes bacteria are difficult to adequately 

sample, identify, and measure. Like cyanobacteria, the actinomycetes may produce geosmin and MIB simultaneously, but 

production is strain specific (Wnorowski, 1992; Watson, 2003; 

Zaitlin and others, 2003; Zaitlin and Watson, 2006). Total 

actinomycetes, rather than identification at the genera or strain 

level, typically are measured. Strain isolation and culture are 

required to conclusively determine actinomycetes production 

of geosmin and/or MIB. 

Detection of Cyanobacterial Toxins and Tasteand-Odor Compounds

A variety of analytical techniques exist to aid in the 

determination of cyanobacterial toxins (Chorus and Bartram, 

1999; Nicholson and Burch, 2001; Fastner and others, 2002; 

Hawkins and others, 2005; Msagati and others, 2006; Nicholson and others, 2007; Osswald and others, 2007). Techniques 

include bioassays such as enzyme-linked immunosorbent 

assays (ELISA) (Metcalf and Codd, 2003), protein phosphatase inhibition assays (PPIA), neurotransmitter inhibition 

assays (acetylcholinesterase inhibition) (Devic and others, 

2002), radioassays (Llewellyn and others, 2001), gas chromatography flame ionization detection (GC-FID), mass spectrometry (GC/MS) (Edwards and others, 1992), liquid chromatography (LC) coupled with ultraviolet-visible spectrometry 

(LC/UV-Vis) (Welker and others, 2002; Barco and others, 

2005), fluorescence (LC/Fluorescence), mass spectrometry 

(LC/MS) (Dell’Aversano and others, 2004), tandem mass 

spectrometry (LC/MS/MS) (Dahlman and others, 2003; Spoof 

and others, 2003), and ion trap mass spectrometry (LC/ITMS). 

Other techniques have been utilized but are not encountered as 

frequently or are not well suited for quantitative work (Chorus 

and Bartram, 1999). 

Relative advantages and disadvantages of common analytical techniques utilized for analysis of cyanobacterial toxins 

and taste-and-odor compounds are shown in table 3. Bioassays 

typically are easy to learn and use, and relatively cost effective when used as a screening tool or, when appropriate, for 

toxicity assessment (Chorus and Bartram, 1999; Msagati and 

others, 2006); however, cross-reactivity can lead to a lack of 

specificity for the target analyte(s) (Metcalf and others, 2002). 

Table 2. Common cyanobacterial toxins, toxicity (based on intraperitoneal mouse assays), and common effects of exposure.

[Most toxin groups have several variants with a range of toxicities. Although known chronic effects are listed, the chronic effects of exposure to cyanotoxins 

currently (2008) are not well understood. LD50, lethal dose required to kill half of the members of a tested population; μg/kg, micrograms per kilogram of body 

weight; -, no data available; >, greater than]

Class Toxin Toxicity 

(LD50) Acute effects Chronic effects

Neurotoxins Anatoxins 20 – 250 μg/kg Seizure, paralysis, respiratory failure, 

death 

unknown

Saxitoxins1 10 μg/kg Tingling or numbness in extremities, 

paralysis, respiratory failure, death 

unknown

β-N-methylamino-L-alanine (BMAA) - - neurodegenerative 

disease

Hepatotoxins Microcystins 25 – > 1,000 μg/kg Acute hepatoenteritis, shortness of 

breath, interhepatic hemorrhage 

hemorrhagic shock, heart failure, 

death

chronic liver injury, 

tumor promoter

Cylindrospermopsins 200 – 2,100 μg/kg Acute hepatoenteritis, renal, lung, 

heart, spleen, thymus, and adrenal 

damage, death

potential 

carcinogen, 

mutagen

Nodularins2 50 μg/kg Similar to microcystins tumor promoter

Dermatoxins Lyngbyatoxins 300 μg/kg Severe dermatitis, gastroenteritis tumor promoter

Aplysiatoxins 300 μg/kg Severe dermatitis, gastroenteritis tumor promoter

Lipopolysaccharides - Dermatitis, gastroenteritis unknown

1

Also known as paralytic shellfish poisons (PSPs).

2

To date, nodularins have only been detected in brackish waters.

 Sources: Chorus and Bartram (1999), Falconer and Humpage (2006), and Stewart and others (2006).
Introduction 9

If toxin-specific information is required, the chromatographic 

techniques are more appropriate, but the cost is greater per 

sample in comparison to bioassays. Researchers are utilizing 

multi-toxin LC/MS/MS methods more frequently because of 

the ability to distinguish a larger variety of individual toxins more readily, and avoid the derivitization that would be 

required for any GC-based technique. LC/MS and LC/MS/MS 

can suffer from matrix effects more than the other techniques, 

but it is possible to compensate for this problem through 

standard addition or isotope dilution (Vogeser and Parhofer, 

2007; Lanckmans and others, 2008). However, isotope dilution 

(Vogeser and Parhofer, 2007) currently (2008) is not an option 

because labeled toxins are not commercially available. 

Fewer analytical options are available for determination 

of taste-and-odor compounds. Closed- and open-loop stripping techniques combined with GC-FID or GC-MS have been 

used previously, but the stripping techniques largely have been 

replaced by solid-phase microextraction (SPME) where phase 

transfer of the semi-volatile compounds to the gas phase is followed by separation and detection by GC/MS. Typical method 

reporting levels range from 1 to 5 nanograms per liter (ng/L), 

the lower threshold for human detection of these compounds 

(Zimmerman and others, 2002; Taylor and others, 2005). Currently (2008), there are no commercially available ELISA’s for 

taste-and-odor analysis sensitive enough to be of practical use.

Table 3. Relative advantages and disadvantages of common analytical techniques utilized for analysis of cyanobacterial toxins and 

taste-and-odor compounds.

Analytical techniques Advantages Disadvantages

Bioassays

 Enzyme-Linked Immunosorbent Assay (ELISA) Relatively easy to use Data interpretation can be difficult

 Inhibition Assays Cost per analyses lowest of 

 all techniques

Inhibition assays and radioassays not 

always available

 Radioassays Can be useful as screening 

 tools

Bioassays frequently possess some 

reactivity towards compounds other 

than the intended target

Can indicate toxicity in some cases Radioassays require permits to work with 

radioisotopes

Research objectives may require a 

chromatographic technique for 

compound specific quantitation

Gas Chromatography (GC)

Flame Ionization Detector (GC/FID) Compound specific Toxins will most likely require 

derivitization

Mass Spectrometry (GC/MS) Cost per analyses intermediate Not all compounds amenable to 

derivitization

Compound identification by GC/MS is 

superior to GC/FID

GC/FID may require further confirmation

Sample concentration techniques may be 

necessary

Liquid Chromatography (LC)

Ultraviolet-Visible (LC/UV-Vis) Derivitization typically not necessary Matrix effects can be substantial

Fluorescence (LC/Fluorescence) Compound specific Cost per sample most expensive 

Mass Spectrometry (LC/MS) Greatest number of toxins are amenable to 

LC techniques

Spectroscopic techniques may require 

further confirmation

Tandem Mass Spectrometry (LC/MS/MS) Cost per analyte can be lowest in a multianalyte methodSample concentration techniques may be 

necessary

Ion Trap Mass Spectrometry (LC/ITMS) Compound identification is superior by 

LC/MS/MS or LC/ITMS
10 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Spatial and Temporal Variability of 

Cyanobacteria and Associated Toxins 

and Taste-and-Odor Compounds

The spatial and temporal variability of cyanobacterial 

populations depends on lake/reservoir hydrology, morphology, 

geography, water chemistry, and biological interactions. Spatiotemporal occurrence patterns are unique to individual lakes, 

and generalizations are difficult because of the large degree of 

variability among lakes. Likewise, a diverse range of physical, chemical, and biological factors may potentially limit 

algal growth, and no one variable is an unequivocal link to 

cyanobacterial bloom formation. Long-term patterns in when, 

where, and under what conditions cyanobacterial blooms typically develop may emerge in lakes that have been studied for 

several years, or cyanobacterial blooms may only occur sporadically (Reynolds, 1984, 1998; Chorus and Bartram, 1999; 

Wetzel, 2001; Falconer, 2005). Local knowledge of general 

lake/reservoir limnology and cyanobacterial bloom occurrence 

will enhance the ability to select appropriate sampling times 

and locations to meet study objectives. If historical data are 

unavailable, initial sampling schedules may be more intensive 

until general patterns in occurrence begin to emerge (Chorus 

and Bartram, 1999; Falconer, 2005). 

Cyanobacterial toxins and taste-and-odor compounds 

occur in two different phases: the particulate phase and the 

dissolved phase. The particulate phase comprises compounds 

that are maintained intracellularly by the cyanobacteria. 

Intracellular toxin and taste-and-odor concentrations are 

strain dependent and may vary by several orders of magnitude between strains. Current (2008) research indicates that 

spatial and temporal variability in toxin and taste-and-odor 

concentrations are largely because of changes in cyanobacterial abundance and strain composition rather than changes in 

intracellular content (Rashash and others, 1996; Orr and Jones, 

1998; Chorus, 2001; Long and others, 2001; Oudra and others, 

2002). The dissolved phase comprises extracellular compounds that have been released into the water column either 

actively by healthy cells or passively upon cell lysis and death. 

Combined, the particulate (intracellular) and dissolved (extracellular) phases comprise total toxin or taste-and-odor concentration (Jones and Korth, 1995; Chorus and Bartram, 1999; 

Graham and Jones, 2007). Particulate, dissolved, and total 

toxin and taste-and-odor compound concentrations may all be 

measured (Graham and Jones, 2007); however, measured total 

and intracellular concentrations are dependent on the efficacy 

of extraction methods (Chorus and Bartram, 1999). 

Cyanobacterial toxins generally are maintained intracellularly until death and cell lysis. Dissolved toxin concentrations 

remain low until populations either begin to senesce naturally 

or are lysed by management practices, such as application of 

algaecides. Because cyanobacterial toxins typically are maintained intracellularly, toxins generally have a similar spatial 

distribution as the cyanobacteria (Chorus and Bartram, 1999; 

Falconer, 2005). Theoretically, the partitioning of cyanobacterial toxins between the particulate and dissolved phases 

will change with time as population density increases and 

declines; particulate concentrations will track overall changes 

in cyanobacterial population density, whereas dissolved 

concentrations will remain relatively low until population 

density begins to decline. The general pattern in the temporal 

distribution of total, particulate, and dissolved cyanobacterial 

toxin concentrations, with respect to cyanobacterial population 

density, is illustrated in figure 4. Field observations indicate 

Relative concentration

Increasing time Not to scale

Cyanobacteria density and

intracellular (particulate) toxin

concentration

Total toxin concentration

Extracellular (dissolved)

toxin concentration Figure 4. Theoretical temporal 

distribution of total, particulate, 

and dissolved cyanobacterial toxin 

concentrations with respect to 

cyanobacterial population density.
Spatial and Temporal Variability of Cyanobacteria and Associated Toxins and Taste-and-Odor Compounds 11

that this pattern occurs in lakes and reservoirs (Jones and 

Orr, 1994; Heresztyn and others, 1997; Chiswell and others, 1999; Chorus and Bartram, 1999); however, many other 

patterns also may occur because toxic and non-toxic strains 

of cyanobacteria can co-occur and will not necessarily have 

the same population dynamics. The partitioning of taste-andodor compounds between the particulate and dissolved phases 

is more complex, because healthy cyanobacterial cells may 

maintain these compounds intracellularly or release them into 

the environment; therefore, taste-and-odor compounds are 

not necessarily associated with the cyanobacteria and may or 

may not have similar spatial distributions (Rashash and others, 

1996; Watson, 2003; Taylor and others, 2005).

The environmental fate and transport of cyanobacterial 

toxins and taste-and-odor compounds under natural lake/reservoir conditions has not been well studied. Available laboratory 

data indicate that photolysis and biodegradation are the most 

likely mechanisms for the degradation of cyanobacterial byproducts in lakes and reservoirs (Kiviranta and others, 1991; 

Stevens and Krieger, 1991; Tsuji and others, 1994; Cousins 

and others, 1996; Chiswell and others, 1999; Holst and others, 

2003; Ou and others, 2005; Kato and others, 2007). Based on 

field observations, dissolved phase toxins and taste-and-odor 

compounds may persist for several days to weeks after the 

decline of a cyanobacterial population (Jones and Orr, 1994; 

Heresztyn and others, 1997; Chorus and Bartram, 1999). The 

persistence of toxins and taste-and-odor compounds, coupled 

with the potential for transport, may lead to spatial or temporal 

uncoupling of cyanobacteria and their by-products. 

Temporal Variability

Seasonal patterns in cyanobacterial abundance and 

community composition are affected substantially by temperature, solar irradiance, and nutrient supply. Cyanobacterial 

populations tend to peak between mid-summer and early fall 

when water temperatures are at seasonal maxima and nutrient 

concentrations are at seasonal minima; however, cyanobacteria 

may remain abundant year round, even in temperate lakes, or 

peak under ice during winter (Reynolds, 1984, 1998; Chorus 

and Bartram, 1999; Wetzel, 2001; Falconer, 2005). Cyanobacterial populations also may vary on much shorter time scales 

(hours or days). Whereas cyanobacterial abundance may 

increase fairly rapidly (days), vertical migration, entrainment 

in temporary circulation cells, or wind movement of surface 

accumulations may change the location of cyanobacteria 

within the lake/reservoir or water column, but not overall 

abundance. Knowledge of local conditions, including patterns 

in circulation, mixing, and prevailing winds, will enhance 

overall study design and aid in the interpretation of observed 

changes in cyanobacterial abundance and community structure 

(Chorus and Bartram, 1999; Falconer, 2005).

Spatial Variability in Lakes and Reservoirs

Cyanobacterial movement affects distribution in the water 

column, and wind movement of surface accumulations may 

have a substantial effect on where cyanobacteria are located 

within a lake or reservoir; however, the general location of 

cyanobacteria largely is affected by lake/reservoir hydrology 

and morphology. Cyanobacterial populations may be evenly 

distributed throughout a lake or reservoir or have an irregular 

distribution because of currents and prevailing winds (fig. 5). 

Statistically significant differences in toxin concentrations 

have been detected within sampling distances of 10 m (Carmichael and Gorham, 1981; Lanaras and others, 1989; Kotak 

and others, 2000), although extreme spatial variation is not 

necessarily typical (Jungmann and others, 1996). Spatial distribution may change rapidly because of changing hydrologic 

conditions such as new circulation patterns or inflow events. 

After heavy rains cyanobacteria in the reservoir shown in figure 5 were washed from up-reservoir areas into the main body 

of the reservoir. The cyanobacterial population in this case was 

restricted to the old river channel (near the tree line). In larger 

lakes or reservoirs certain areas (shallow bays and coves, sites 

directly affected by nutrient rich inflows, or structures that 

affect flow such as dikes, piers, or potentially larger intake 

towers) may be more prone to the development of cyanobacterial blooms and surface accumulations. These isolated blooms 

may remain localized or spread throughout the lake because of 

inflow events or circulation patterns (Reynolds, 1984; Chorus and Bartram, 1999; Wetzel, 2001; Falconer, 2005). Some 

knowledge of the distribution of cyanobacteria, as affected by 

hydrology, morphology, and typical conditions, throughout 

a lake is required before the development of toxin and tasteand-odor studies. The sites selected for study will depend 

on the overall study objectives (Chorus and Bartram, 1999; 

Falconer, 2005). 

Spatial Variability in the Water Column

The ability of many toxin and taste-and-odor producing 

cyanobacteria to control their position in the water column 

needs special consideration when sampling for these compounds. The vertical distribution of cyanobacteria may vary 

widely for relatively short (hours or days) periods of time 

(Reynolds, 1984; Chorus and Bartram, 1999; Falconer, 2005). 

Sample location in the water column relative to cyanobacterial distribution may have a substantial affect on results. For 

example, if cyanobacteria are maintaining a position at depth 

in the water column, and surface samples are collected, toxins 

or taste-and-odor compounds may not be detected even when 

present. There are six general water column distributions of 

cyanobacteria; detailed descriptions and occurrence information are given in figures 6 and 7:
12 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

1. Shoreline, near-shore, or open-water surface accumulations and scums (fig. 6A and fig. 7A). Surface accumulations and scums are of greatest concern for recreational 

exposure to high concentrations of cyanobacterial toxins.

2. Even distribution throughout the photic zone or epilimnion (fig. 6B and fig. 7B).

3. Specific depth in the photic zone (fig. 6C and fig 7C). In 

shallow lakes where light penetrates to the bottom (photic 

depth equals maximum depth), cyanobacteria may be 

located on or near the bottom.

4. Metalimnetic bloom (fig. 6D, fig. 7D, fig. 8). This is a 

special case of a population maintaining a specific depth 

in the photic zone. Metalimnetic blooms are evident by 

sharp increases in oxygen at depth, typically near the 

interface of the epilimnion and metalimnion (figs. 1 

and 8). Metalimnetic blooms are of particular concern in 

drinking-water supplies because the populations are not 

visibly evident and may be located at the same depth as 

the drinking-water intake.

5. Even distribution throughout the water column (fig. 6E

and fig. 7E).

6. Under ice (fig. 6F and fig. 7F).

The distribution of cyanobacteria in the water column 

often can be determined by a combination of visual assessment, light profiles to determine photic depth, and temperature 

and dissolved oxygen profiles to determine stratification, 

mixed depth, and presence of a metalimnetic population 

(figs. 6–8). Cyanobacteria often will maintain a position in the 

photic zone, regardless of mixed depth (Reynolds and Walsby, 

1975; Reynolds, 1987; Chorus and Bartram, 1999; Wetzel, 

2001; Falconer, 2005; Graham and others, 2006); therefore, 

if cyanobacterial distribution in the water column cannot be 

conclusively determined, an integrated photic zone sample 

will likely be fairly representative.

Once toxins and taste-and-odor compounds are released 

by cyanobacteria into the dissolved phase, they will become 

uniformly distributed throughout the mixed layer in which 

they are released; this typically is a rapid process, but large 

localized concentrations may develop in hydraulically isolated areas such as coves (Jones and Orr, 1994). Generally, 

Figure 5. Example of spatial variability of cyanobacteria within a reservoir.
Study Types, Objectives, Designs, and Approaches for Studies of Cyanobacteria, Toxins, and Taste-and-Odor Compounds 13

the distribution of dissolved compounds in the water column 

will depend on the layer in which the cyanobacteria were 

located when the compounds were released. In stratified lakes, 

cyanobacteria most likely are in the epilimnion or, less frequently, the metalimnion. Dissolved toxin and taste-and-odor 

samples usually can be collected from the same water column 

location as samples for intracellular (particulate) analysis 

(Chorus and Bartram, 1999; Falconer, 2005).

Common Study Types, Objectives, 

Designs, and Approaches for Studies of 

Cyanobacteria and Associated Toxins 

and Taste-and-Odor Compounds

Averett and Schroder (1994) describe three general 

types of surface-water-quality studies: reconnaissance studies, monitoring studies, and interpretive studies. These study 

types do not have well-defined boundaries, and often there 

is overlap between types as study objectives and designs are 

developed. Cyanobacterial toxin and taste-and-odor studies 

are discussed with respect to these three study types. The keys 

to a well designed study include using the scientific method, 

a well-defined problem, clear objectives, and an appropriate 

approach. Study objectives will dictate what variables need 

to be measured, and when, where, and how samples will be 

collected. More detail on the general types of surface-waterquality studies and the fundamentals of study design are given 

in Averett and Schroder (1994). 

The study of cyanobacterial toxins and taste-and-odor 

compounds is an active area of developing research, but 

many fundamental questions remain about the occurrence, 

environmental causes, and ecological consequences of these 

compounds (Chorus, 2001). Current (2008) studies include: 

reconnaissance studies to evaluate the occurrence, distribution, 

and concentration of cyanobacterial toxins and taste-and-odor 

compounds; monitoring studies to determine the potential for 

human health risks and taste-and-odor events associated with 

cyanobacterial toxins and taste-and-odor compounds; and 

a range of interpretive studies. Interpretive studies include: 

evaluation of the physical, chemical, and biological factors 

affecting cyanobacterial and taste-and-odor compound occurrence and concentration and the development of empirical 

models; determination of the environmental fate and transport 

(including persistence, degradation, and movement in the environment and among ecosystem compartments) of cyanobacterial toxins and taste-and-odor compounds; and assessment of 

the toxicological effects of cyanobacterial toxins and tasteand-odor compounds on aquatic and terrestrial organisms and 

overall ecosystem function and health. There are many types 

of studies that can be conducted within these general topical 

A

A

B

C

D

E

F

B E

F

C D

Ice

EXPLANATION

Shoreline, near-shore, and open water accumulations and scums

Even distribution throughout the photic zone or epilimnion

Specific depth in the photic zone

Metalimnetic bloom (special case of C)

Even distribution throughout the water column

Under ice bloom

Potential water column distributions of cyanobacteria

Lake bottom

Even cyanobacteria

distribution Cyanobacteria

accumulation

Not to scale Schematic after Chorus and Bartram (1999)

Figure 6. Potential water column distributions of cyanobacteria.
14 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Appearance Location

Shoreline, near-shore and

open-water surface

accumulations and scums.

Dispersed in photic zone.

Dispersed in epilimnion.

Appearance will vary

depending on the type

and abundance of

cyanobacteria.

Description

Discoloration of the

water surface; typically

blue-green or bright

green but may also be

brown or red1

.

May be a thin layer

near the surface or be

thick or paint-like.

Accumulations may

extend from the

surface to depths of

one meter or more.

Dissolved oxygen may

be supersaturated near

the surface.

Water likely has an

obvious color; typically

blue-green or bright

green but may also be

brown or red.

Large colonies and 

filaments may be

visible

Occurrence

Develop when existing

cyanobacterial

population accumulates

near the water surface.

May occur in open-water

or near-shore areas.

May rapidly move or

dissipate with changes 

in wind speed and direction.

May occur during calm

periods or during the

decline of a cyanobacterial

population.

May be widespread or

have a patchy distribution

depending on wind and 

circulation patterns.

Relatively calm conditions.

May occur in well-mixed

or stably stratified water

column.

Water may or may not 

have obvious color;

typically blue-green or

bright green but may

also be brown or red.

Large colonies or

filaments may or may

not be visible.

Stably stratified water

column.

Breezy, windy conditions

A.

B.

1

From a distance duckweed (Lemna) may be confused with a cyanobacterial bloom; duckweed has a bright green color and may

cover large areas. Duckweed is a plant with visible leaves and up close duckweed is easily distinguished from cyanobacteria.

Duckweed typically proliferates in shallow, nutrient rich ponds, bays, and small lakes/reservoirs. Examples of duckweed appearance

are available on the internet at http://www.mobot.org/jwcross/duckweed/duckpix.htm and http://aquaplant.tamu.edu/database/

floating_plants/common_duckweed_pics.htm.

All photographs by J.L. Graham, USGS Lawrence, Kansas, with the exception of : E., courtesy of

Kansas Department of Health and Environment, and F., courtesy of an anonymous photographer. 

Figure 7. Appearance, location, description, and occurrence of the potential water column distributions of cyanobacteria.
Study Types, Objectives, Designs, and Approaches for Studies of Cyanobacteria, Toxins, and Taste-and-Odor Compounds 15

Appearance Location

Metalimnetic bloom

(special case of a population

maintaining a specific depth

in the photic zone)

Metalimnetic blooms are not

visible at the water surface;

they can be detected by

sharp increase in dissolved

oxygen at depth.

Even distribution throughout

the water column.

Appearance will vary

depending on the type and

abundance of cyanobacteria.

Under ice bloom

Description

Not visible from the

surface.

Evident by sharp increase

in dissolved oxygen at

depth, typically near the

interface of the epilimnion

and metalimnion

Water may or may not

have obvious color;

typically blue-green or

bright green but may

also be brown or red.

Large colonies or

filaments may or may

not be visible

Occurrence

Stably stratified water

column

Intermediate trophic status

(mesotrophic)

Photic zone penetrates

into the metalimnion

Well-mixed conditions in:

 Shallow lakes/reservoirs

 Deeper lakes/reservoirs 

 that do not typically

 stratify

 

Autumn, winter, and spring

 conditions in temperate

 lakes/reservoirs that

 stratify

May be visible under

ice depending on ice

thickness; color

typically blue-green or

red.

Typically visible when

ice plug is removed.

Ice clarity and thickness

allows adequate light

penetration

Specific depth in photic

zone.

Appearance will vary. Here

the cyanobacteria were

near the water surface

during early morning.

Water may or may not 

have obvious color.

Large colonies and

filaments may or may

not be visible

Specific location in the 

photic zone may be

difficult to determine 

without discrete samples.

Relatively calm conditions.

May occur in well-mixed

or stably stratified water

column.

Caution: In shallow lakes where

light penetrates to the bottom (photic

depth equals maximum depth)

cyanobacteria may be located on or

near the bottom. 

C.

D.

E.

F.

All photographs by J.L. Graham, USGS Lawrence, Kansas, with the exception of : E., courtesy of

Kansas Department of Health and Environment, and F., courtesy of an anonymous photographer. 

Figure 7. Appearance, location, description, and occurrence of the potential water column distributions of cyanobacteria.—Continued
16 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

areas. This section is not intended to provide a comprehensive 

list of all possible types of studies and research objectives for 

lake or reservoir studies on toxins and taste-and-odor compounds; rather, it is intended to illustrate how the information 

presented on the spatial and temporal variability of cyanobacteria and cyanobacterial blooms can be used to guide and 

enhance the design of field studies. The examples presented 

cover many, but not all, of the possible objectives, sampling 

frequencies, site locations, and collection methods. 

General study objectives and guidelines on when, where, 

and how to sample are presented for reconnaissance, monitoring, and interpretive studies. Special consideration needs to 

be given to the distribution of cyanobacteria in the study lake 

or reservoir and water column when deciding where and how 

to collect samples; sample location needs to be relevant to 

where the cyanobacterial community is located. Ultimately, 

sampling approach and site selection will depend on specific 

study objectives. Sampling frequency, site location, time of 

day samples are collected, and type of samples collected need 

to be carefully considered with respect to study objectives.

Reconnaissance Studies

Reconnaissance studies typically are conducted to assess 

the occurrence and concentration of cyanobacteria and associated toxins and taste-and-odor compounds. Studies may focus 

on spatial and/or temporal variability at a range of scales, 

from single systems to states or regions. Studies of lakes or 

reservoirs used for recreation typically emphasize cyanobacterial toxins, whereas studies of drinking-water supply lakes 

or reservoirs may emphasize taste-and-odor compounds and 

toxins. General study objectives and guidelines describing 

when, where, and how samples typically are collected for 

reconnaissance studies are presented in table 4. An example of 

a study design, approach, and field form for a reconnaissance 

study is given in Appendix 1.

Monitoring Studies for Public Health and 

Drinking Water Treatment

Monitoring studies typically are conducted to determine 

the concentration of cyanobacterial toxins and taste-and-odor 

compounds in areas where exposure is most likely to occur. 

Recreational areas are monitored for cyanobacterial toxins 

to assess human health risks; results often are used to make 

decisions about posting warnings and closing recreational 

areas. Drinking-water intakes are monitored for taste-and-odor 

compounds and/or toxins to assess the potential for taste-andodor events and human health risks; results often are used to 

guide drinking-water treatment processes. Guidelines describing when, where, and how samples typically are collected for 

monitoring studies are presented in table 5. An example of a 

study design, approach, and field form for a monitoring study 

is given in Appendix 2.

Interpretive Studies

Interpretive studies typically are conducted to 

assess the processes that affect the spatial and temporal 

Surface

Bottom

Not to scale

Temperature

Oxygen

Increasing depth

Increasing values

Increasing values

Figure 8. Graphical representation of the temperature 

and oxygen gradients that 

will develop in the water 

column when a metalimnetic 

cyanobacterial bloom 

is present.
General Guidelines for Sampling 17

distribution and abundance of cyanobacterial toxins and tasteand-odor compounds. Interpretive studies may include, but 

are not limited to, the assessment of the physical, chemical, 

and biological factors affecting occurrence and concentration, 

environmental fate and transport studies, and toxicological 

studies. Studies assessing the environmental factors affecting 

occurrence and concentration often are focused on developing empirical models (Mau and others, 2004; Christensen and 

others, 2006). Several years of study, with a range of hydrologic and meteorologic conditions, often are required before 

true patterns begin to emerge. Long-term studies are required 

to develop methods for early detection, such as real-time estimation of the occurrence or concentration of cyanobacterial 

toxins and taste-and-odor compounds, and models predicting 

when events are likely to occur. Reliable tools for early detection and predictive models will allow resource managers time 

to respond more effectively to potentially harmful conditions. 

General study objectives and guidelines describing when, 

where, and how samples typically are collected for studies 

assessing the environmental factors affecting the occurrence 

and concentration of cyanobacterial by-products are presented 

in table 6. An example of a study design, approach, and field 

form for an interpretive study is given in Appendix 3.

Fate and transport and toxicological studies are crucial to 

the overall understanding of environmental and public health 

risks associated with cyanobacterial toxins and taste-and-odor 

compounds. These studies frequently collect water-column 

samples from lakes and reservoirs; they often also require 

samples from sediments, benthic organisms, and tissues. Few 

data are available regarding the partitioning of cyanobacterial toxins and taste-and-odor compounds in these alternative 

ecosystem compartments; sampling protocols and analytical 

methods either have not been developed or currently (2008) 

are under development. Discussion of how to collect these 

types of samples is beyond the scope of this chapter and 

examples are not provided for fate and transport and toxicological studies. 

General Guidelines for Sampling

Three general types of water-quality samples are collected from lakes and reservoirs: surface samples, discretedepth samples, and depth-integrated samples. Depth-integrated 

samples may encompass the photic zone, epilimnion, metalimnion, hypolimnion, entire water column, or other layers of 

interest depending on study objectives. The types of samplers 

commonly used to collect each type of sample are listed in 

table 7. Detailed descriptions of the samplers and proper 

uses are presented in Britton and Greeson (1987), the USGS 

National Field Manual (NFM) chapters 2 (Lane and others, 

2003) and 4 (U.S. Geological Survey, 2006), and Standard 

Methods Sections 1060 and 10200 (Clesceri and others, 

1998). Because toxins and taste-and-odor compounds are 

organic compounds, the samplers and churns used to collect 

and composite samples need to be made of fluorocarbon polymers such as Teflon (Lane and others, 2003).

Some cyanobacterial colonies or filaments may exceed 

the maximum particle size (250 micrometers) recommended 

for use of a churn splitter (Lane and others, 2003); however, 

particle size does not matter when dealing with algae. There 

are no settling issues with algae like there are with sediments. 

Churning velocity will maintain all algal size classes within 

the sample. Likewise, churning velocity also will maintain all 

cyanobacterial size classes within a sample regardless of buoyancy. Although some colonies and filaments may break apart 

during the churning process, individual cell integrity should 

not be affected by churning. Use of churns is inappropriate 

when collecting samples from thick accumulations with little 

water content.

For each sample type, a single grab sample may be collected or multiple grab samples may be composited into a 

churn. Whether a single grab or composite sample is collected 

will depend on study objectives and the volume of water 

needed. How samples are composited also will depend on 

study objectives. If specific locations are of interest, composites may be collected from the same depth and location until 

the required volume of water has been acquired; if general 

or average conditions are of interest, composites may be 

composed of samples collected from multiple depths in the 

water column or multiple locations in the lake or reservoir. 

Composite samples are more representative of overall conditions; however, because of the generally irregular distribution of phytoplankton communities, concentrations of toxins 

and taste-and-odor compounds may be diluted (Clesceri and 

others, 1998). 

Surface Samples

If surface samples are collected in open-water areas, 

samples need to be collected 0.5–1.0 m below the surface to 

avoid substances that are trapped in the surface film and may 

affect results (Clesceri and others, 1998). If surface accumulations or scums are being sampled, then collection at the water 

surface is appropriate. Study objectives need to be considered 

when determining how to sample surface accumulations and 

scums because cyanobacterial density may vary widely across 

the area of accumulation. If a general idea of maximum toxin 

and/or taste-and-odor compound concentration is desired, 

a sample collected from the thickest part of the accumulation or scum is adequate (Chorus and Bartram, 1999). If an 

average concentration in the accumulation or scum is desired, 

multiple samples can be collected throughout the area and 

composited. A grid or transect approach often is used for this 

type of collection (Clesceri and others, 1998). The number of 

samples used for a composite will depend on study objectives 

and the size of the accumulation or scum. If information on 

spatial heterogeneity is desired, samples need to be collected 

throughout the accumulation or scum area, but not composited 

(Clesceri and others, 1998; Chorus and Bartram, 1999).
18 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Discrete-Depth Samples

Discrete-depth samples typically are collected when the 

location of the cyanobacterial community is known, there 

is a structure of interest at depth (such as a drinking water 

intake), or when the vertical water column distribution of the 

cyanobacterial community and associated toxins and tasteand-odor compounds is of interest.

Depth-Integrated Samples

Depth-integrated samples typically are collected when the 

cyanobacterial community is evenly distributed throughout a 

layer of interest, or the interest is in dissolved concentrations 

only. An exception may be when the distribution of cyanobacteria in the water column can not be determined; in this case 

an integrated photic zone sample will be generally representative. There are two types of depth-integrated samples: continuous and discontinuous. Continuous samples incorporate the 

entire depth of interest and are collected using pumps or bailers. Several continuous samples may be composited to obtain 

the necessary sample volume. Discontinuous samples do not 

incorporate the entire depth of interest, and are collected using 

Kemmerer or Van Dorn samplers, although the percent of the 

water column represented can be increased by using a vertical sampler rather than a horizontal sampler (table 7; Lane 

and others, 2003; U.S. Geological Survey, 2006). A depthintegrated sample may be obtained by collecting and compositing several discontinuous samples from several depths within 

the layer of interest. If discontinuous samples are collected, 

sample intervals need to be equal (for example every meter 

within the photic zone) (Chorus and Bartram, 1999; Clesceri 

and others, 1998). 

Quality Control

Quality-control samples, such as blanks, splits, and replicates, are an integral component of well designed water-quality studies and are used to assess variability caused by sample 

collection, processing, shipping, and handling. Quality-assurance plans and the types of quality-control samples collected 

will depend on study design and objectives. Detailed information on how to develop quality-assurance plans and descriptions of the different types of quality-control samples are 

discussed elsewhere (Friedman and Erdmann, 1982; Shampine 

and others, 1992; U.S. Geological Survey, 2006).

Cyanobacterial communities tend to have an irregular 

distribution within the water column and throughout a lake or 

reservoir; therefore, concurrent and sequential field replicates 

(U.S. Geological Survey, 2006) to assess sampling variability 

are particularly important in studies of cyanobacterial toxins 

and taste-and-odor compounds. Methods for the extraction and 

analysis of toxins and taste-and-odor compounds are evolving 

rapidly and currently (2008) are not consistent among laboratories. Split samples (U.S. Geological Survey, 2006) to assess 

intra- and inter-laboratory variability in extraction and analysis 

are useful. 

Sample Handling, Preparation, Processing, and 

Shipping for Analyses

There are several cyanobacterial toxins and taste-andodor compounds that commonly are measured including the 

toxins microcystin, cylindrospermopsin, anatoxin, and saxitoxin and the taste-and-odor compounds geosmin and MIB. 

BMAA is not characterized well enough at this time to provide 

guidelines on sampling procedures or sample handling. A key 

decision when designing toxin and taste-and-odor studies is 

whether total, particulate, or dissolved concentrations will be 

analyzed. Sample handling, preparation, and processing will 

partly be determined by whether total, particulate, and/or dissolved compounds are measured. Another important determinant is the instructions provided by the laboratory conducting 

the analyses. Laboratories may request whole water samples 

and do all processing in house; others will provide specific 

instructions including the type of sample bottle to use, and 

yet others will provide instructions as well as sample bottles 

and preservatives. There also may be restrictions on delivery 

days that may affect analyses on unpreserved samples. Clear 

communication with the laboratories performing the analyses 

is required to ensure sample analyses will provide the desired 

results. Although handling, preparation, processing, and shipping largely may be determined by the analyzing laboratory, 

general guidelines are provided below. 

Sample Handling

Ideally, all samples are processed on site. If samples need 

to be transported before processing, they are placed on ice 

in the dark because heat and light may cause degradation of 

cyanobacterial cell integrity, toxins, and taste-and-odor compounds. All samples need to be processed the same day they 

are collected. 

Many cyanobacterial toxins do not degrade when frozen, 

and samples may be stored in a freezer for several months or 

years (Chorus and Bartram, 1999). Keep in mind that if whole 

water samples are frozen, cyanobacterial cells will lyse and 

release intracellular toxins; freezing and thawing whole water 

samples frequently is used to determine total toxin concentration (Gjølme and Utkilen, 1994; Chorus and Bartram, 1999; 

Johnston and Jacoby, 2003; Graham and Jones, 2007). Tasteand-odor samples generally are not frozen. 

Processing Toxin and Taste-and-Odor Samples

Total Concentration Samples—Total samples measure 

both particulate (intracellular) and dissolved (extracellular) phases of cyanobacterial toxins and/or taste-and-odor 

compounds. Total concentrations are measured directly by 
General Guidelines for Sampling 19

Table 4. General study objectives and guidelines describing when, where, and how samples typically are collected for 

reconnaissance studies.

General objective Site location Sample frequency Sample type

Regional studies

Spatial variability

Emphasis on presence/absence Single representative site, typically an open, deep water siteSingle point in time when most

cyanobacterial-related issues

occur

Integrated photic zone

Integrated epilimnion

Surface sample

Site will be determined based

on the location of surface

accumulations and scums

During known surface bloom

events

Surface sample

Spatial and temporal variability

Emphasis on presence/absence

and changes in concentration

with time

Single representative site, typically an open, deep water siteMultiple times during the period

when most cyanobacterialrelated issues occur

•	Weekly

•	Bi-weekly

•	Monthly

•	Annually

Integrated photic zone

Integrated epilimnion

Surface sample

Single-system studies

Spatial variability

Emphasis on presence/absence Multiple sites Single point in time when a

cyanobacterial bloom is 

occurring

Integrated photic zone

Integrated epilimnion

Integrated water column

Surface sample

Spatial and temporal variability

Emphasis on presence/absence

and changes in concentration

over time

Multiple sites Multiple times during the period

when most cyanobacterialrelated issues occur

•	Weekly

•	Bi-weekly

•	Monthly

Integrated photic zone

Integrated epilimnion

Integrated water column

Surface sample

Emphasis on spatial changes 

within the lake or water 

column over relatively short 

periods of time

Single representative site

Multiple sites

Multiple points in time when a 

cyanobacterial bloom is 

occurring

•	Hourly

•	Daily

Integrated photic zone

Integrated epilimnion

Integrated water column

Surface sample

Discrete depth

Sources: Chorus and Bartram (1999), Falconer (2005), and Wetzel and Likens (2000).
20 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Table 5. General study objectives and guidelines describing when, where, and how samples typically are collected for monitoring 

studies. 

General objective Site location Sample frequency Sample type

Recreational areas Beaches

Open water areas used for fullbody contact recreation

Bay or cove areas used for fullbody contact recreation

Public access sites

Routine basis during periods of 

peak recreational use

•	Daily

•	Weekly

Surface sample

Integrated photic zone

Drinking-water supplies Location relevant to the drinkingwater intake(s)Routine basis

•	Daily

•	Weekly

During periods when events have 

historically occurred

During events

Discrete depth

Integrated photic zone

Integrated epilimnion

Integrated water column

Sources: Chorus and Bartram (1999), Falconer (2005), and Wetzel and Likens (2000).

Table 6. General study objectives and guidelines describing when, where, and how samples typically are collected for interpretive 

studies assessing the physical, chemical, and biological factors affecting the occurrence and concentration of cyanobacteria and 

associated toxins and taste-and-odor compounds.

General objective Site location Sample frequency Sample type

Environmental factors 

influencing spatial and/or 

temporal occurrence 

Real-time estimation of 

occurrence/concentration

Predictive models

Single representative site, 

typically an open, deep 

water site

•	Sites for drinking-water 

 studies are typically 

 located near intakes

Multiple sites

•	Sites where cyanobacterial 

 blooms are known to 

 initiate

•	Sites where cyanobacteria 

 are typically abundant

•	Inflow sites1

Routine basis

•	Weekly

•	Bi-weekly

•	Monthly

Integrated photic zone

Integrated epilimnion

Integrated water column

Discrete depth

Sites where surface accumulations/scums are locatedEvent samples

Sampling plans need to be 

flexible enough to respond to 

events

Surface sample

Sources: Chorus and Bartram (1999), Christensen and others (2006), Falconer (2005), and Wetzel and Likens (2000).

1

Monitoring of major inflows is essential in the development of predictive models and beneficial in the development of models for real-time estimation of 

occurrence/concentration. Standard USGS protocols for sampling streams and rivers should be used when sampling inflows (U.S. Geological Survey, 2006). 

Cyanobacterial samples and samples for toxin and taste-and-odor analysis may or may not be analyzed at inflow sites, depending on conditions; however, 

actinomycetes samples need to be collected.
Safety Considerations 21

rupturing cyanobacterial cells and releasing the compounds 

into solution (note that total concentrations are dependent 

on the efficacy of extraction techniques) (fig. 9). Whole 

water samples are typically collected for total concentration 

analysis in 500- to 1,000-mL bottles, commonly high-density 

polyethylene (HDPE) plastic or amber glass. Bottle type and 

volume will depend on the laboratory doing the analysis. Total 

concentrations also may be measured indirectly by summing 

particulate and dissolved concentrations (Chorus and Bartram, 

1999; Graham and Jones, 2007).

Particulate Concentration Samples—Particulate samples 

measure only the intracellular phase of cyanobacterial toxins 

and/or taste-and-odor compounds. Particulate concentrations 

are measured directly by concentrating cyanobacteria from 

a known volume of water onto a filter (fig. 9) (Chorus and 

Bartram, 1999). The filter is extracted prior to analysis (note 

that particulate concentrations are dependent on the efficacy 

of extraction techniques). Samples for particulate analysis are 

collected on 0.7-micrometer (μm) glass fiber filters, and processed in the same manner as chlorophyll samples; processes 

are described in NFM chapter 7.4 (Hambrook Berkman and 

Canova, 2007). Because toxins and taste-and-odor compounds 

are organic compounds, glass or metal filter funnels and glass 

receiving flasks need to be used. When filtering, remember to 

accurately measure and record the filtered volume. Particulate 

concentrations also may be measured indirectly by taking the 

difference between total and dissolved concentrations (Chorus 

and Bartram, 1999; Graham and Jones, 2007).

Dissolved Concentration Samples—Dissolved samples 

measure only the extracellular phase of cyanobacterial toxins 

and/or taste-and-odor compounds. Filtrate from 0.7-μm glass 

fiber filters is collected for dissolved toxin analysis (fig. 9). A 

glass or metal filter funnel and glass receiving flask are used 

for the filtration process. Filtrate from preparation of particulate toxin samples or chlorophyll samples may be used. Again, 

HDPE plastic and amber glass bottles are typically used (Chorus and Bartram, 1999; Graham and Jones, 2007). Bottle type 

and volume will depend on the laboratory doing the analysis. 

Analysis of taste-and-odor compounds most commonly is 

performed on dissolved samples, but total and particulate concentrations also are determined regularly (van der Ploeg and 

others, 1992; Wnorowski and Scott, 1992; Jones and Korth, 

1995; Yen and others, 2007).

Toxin and taste-and-odor concentrations typically are 

expressed volumetrically as micrograms per liter (μg/L) 

because volumetric concentrations are easily related to drinking water and recreational guideline values. Concentrations 

also may be expressed gravimetrically as micrograms per 

gram dry weight (μg/g dry weight). To estimate dry weight, an 

additional sample can be prepared as for particulate analysis 

and weighed after desiccation or lyophilization. Particulate 

concentrations may then be expressed either volumetrically or 

gravimetrically. Gravimetric concentrations need to be interpreted with caution. When collected from dense accumulations 

of cyanobacteria, gravimetric expression of toxin or taste-andodor concentration often is directly related to cyanobacterial 

abundance; however, when collected in open-water areas, dry 

weight will incorporate all suspended particulates including 

other phytoplankton, zooplankton, and sediment (Chorus and 

Bartram, 1999).

Processing Cyanobacterial (Phytoplankton) 

Samples

Whole water samples are collected for cyanobacterial 

community composition analysis, abundance (cells or natural units per milliliter or liter), and biovolume (micrometers 

cubed per milliliter or liter) (Blomqvist and Herlitz, 1998; 

Olrik and others, 1998). Bottle type, volume, and preservative 

used depend on the laboratory doing the analysis. Because 

there can be great variability among analysts, it is important 

to confirm counting and processing methodologies before the 

analysis, including subsample preparation, counting threshold, 

taxonomic references, and the experience of the analyst. This 

is particularly important when using multiple laboratories 

for analysis. Cyanobacteria are considered to be part of the 

phytoplankton community, and there is no difference between 

samples collected for cyanobacterial analysis and phytoplankton analysis. More detail on how to collect and process 

phytoplankton samples can be found in Hambrook Berkman 

and Canova (2007).

Sample Shipping

Samples need to be kept chilled (on ice or refrigerated) 

and in the dark until shipped. Ideally, samples are shipped 

on the same day they are collected following the instructions 

Table 7. Samplers commonly used to collect surface, discrete 

depth, and depth integrated samples in lakes and reservoirs. 

[Descriptive information on the samplers as well as appropriate use is available in Britton and Greeson, 1987, U.S. Geological Survey National Field 

Manual chapters 2 (Lane and others, 2003) and 4 (U.S. Geological Survey, 

2006), and Standard Methods Sections 1060 and 10200 (Clesceri and others, 1998).]

Sample type Sampler

Surface Hand-held open mouth bottle sampler

Weighted bottle sampler (US WBH-96)

Discrete depth Kemmerer Bottle

Van Dorn Bottle—horizontal or vertical

Pump—diaphragm or peristaltic

Depth integrated Kemmerer Bottle

Van Dorn Bottle—vertical

Pump—diaphragm or peristaltic

Bailer
22 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

provided by the analyzing laboratory. Typically, samples 

are placed in a cooler, packed with ice (double bagged), and 

shipped using priority overnight mail so samples arrive at the 

analyzing laboratory the next morning. Be aware of sample 

holding times when shipping to areas of the country where 

there is extreme weather. It is better to hold samples under 

controlled conditions than to have samples exposed to extreme 

temperatures for 24 to 48 hours if shipments get delayed. 

Commonly Collected Ancillary Data

The data and additional variables collected during 

cyanobacterial toxin and taste-and-odor studies will depend 

entirely on the objectives of the study. Commonly measured 

variables are listed in table 8. Generally, all sub-samples for 

laboratory analyses, including toxin and taste-and-odor analyses, are collected from the same grab or composite sample. 

The volume of the grab or composite sample needs to be sufficient for all planned analyses.

Safety Considerations

Cyanobacterial toxins are known to cause human illness, and skin contact may result in irritation and rash. 

Gloves always need to be worn when collecting samples for 

cyanobacterial toxin and taste-and-odor analysis. Skin and eye 

contact with dense surface accumulations need to be avoided. 

If dense surface accumulations are going to be sampled, gloves 

that extend to the shoulder may be required. If contact with 

dense surface accumulations occurs, the affected area needs to 

be rinsed immediately with clean water. Because inhalation of 

aerosols may be a problem for those with respiratory illness, 

precautions also need to be taken if sampling personnel have a 

recent history of asthma or respiratory disease. Other general 

safety considerations are discussed in NFM chapter 9 (Lane 

and Fay, 1997).

U.S. Geological Survey National Water 

Information System Parameter Codes 

Many of the cyanobacterial toxin and taste-and-odor 

compounds already have parameter codes available for entry 

into the USGS National Water Information System (NWIS) 

and updates are made as new compounds and analyses become 

available. Check the NWIS parameter code dictionary to see 

what codes are available for toxin and taste-and-odor compounds. The parameter code dictionary is available online at: 

http://waterdata.usgs.gov/nwis/help?codes_help #Table8.

Summary

The study of cyanobacterial toxins and taste-and-odor 

compounds is an active area of developing research. Consistent guidelines for the development of scientific studies 

of cyanobacteria and associated toxins and taste-and-odor 

compounds will allow for nationally comparable data sets and 

among study comparisons. Study objectives and design dictate 

when, where, and how samples are collected for analysis 

of cyanobacteria and associated toxins and taste-and-odor 

compounds and require careful consideration with respect to 

the unique characteristics of the cyanobacteria. This report 

Total

(Intracellular + Extracellular)

Dissolved

(Extracellular) 

Dissolved

+ Toxin/

C

T

o

a

m

st

p

eoa

u

n

n

ddOdor =

DDisisssoollvveedd

Compound

Intracellular

Compound

Particulate

(Intracellular)

Filter with concentrated

cyanobacteria

Filtered water sample Whole water sample

Figure 9. Partitioning of cyanobacterial toxins and taste-and-odor compounds among the total, particulate (intracellular), and 

dissolved (extracellular) phases in different types of samples.
Summary 23

Table 8. Commonly collected ancillary data for studies of cyanobacterial toxins and taste-and-odor compounds and references for 

additional information. 

[Table modified after Table 7.4-4 (Hambrook Berkman and Canova, 2007) in the U.S. Geological Survey National Field Manual (NFM); GPS, global 

positioning system]

Ancillary data Description Reference(s)

Observational data General field observations such as water color and clarity, 

current meteorological conditions, presence of visible algae, 

presence of surface accumulations or scums of cyanobacteria

NFM 4 (U.S. Geological Survey, 2006)

Wetzel and Likens, 2000

Meteorological conditions several days before sampling

Occurrence of recent inflow events

Water residence time

Lake levels

Field measurements Photographs of current conditions NFM 4 (U.S. Geological Survey, 2006) 

NFM 6 (Wilde, variously dated)

NFM 7.4 (Hambrook Berkman and Canova, 2007)

Wetzel and Likens, 2000

GPS coordinates

Vertical profiles of light, temperature, dissolved oxygen, 

specific conductance, pH, turbidity, and in situ fluorescence

Secchi depth

Laboratory analyses Phytoplankton community composition, abundance, and 

biovolume

Procedures described in this report and 

NFM 7.4 (Hambrook Berkman and Canova, 

2007)

Chlorophyll NFM 7.4 (Hambrook Berkman and Canova, 2007)

Nutrients—total nitrogen and phosphorus, total Kjeldahl 

nitrogen (ammonia plus 

organic nitrogen), nitrate, nitrite, 

ammonia, and orthophosphorus

NFM 4.0 (U.S. Geological Survey, 2006)

NFM 4.1 (U.S. Geological Survey, 2006)

NFM 5 (Wilde and others, 2004)

Suspended sediment NFM 4.0 (U.S. Geological Survey, 2006)

NFM 4.1 (U.S. Geological Survey, 2006)

NFM 5 (Wilde and others, 2004)

Actinomycetes bacteria (taste-and-odor studies) NFM 7.1 (Myers and others, 2007)

Alkalinity NFM 4. 0 (U.S. Geological Survey, 2006)

NFM 4.1 (U.S. Geological Survey, 2006)

NFM 5 (Wilde and others, 2004)

NFM 6.6 (Rounds, 2006)

Cations and anions NFM 4.0 (U.S. Geological Survey, 2006)

NFM 4.1 (U.S. Geological Survey, 2006)

NFM 5 (Wilde and others, 2004)

Carbon—total, dissolved, and particulate NFM 4.0 (U.S. Geological Survey, 2006)

NFM 4.1 (U.S. Geological Survey, 2006)

NFM 5 (Wilde and others, 2004)
24 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

provides general background information on, as well as guidance for, the design of studies assessing cyanobacterial toxin 

and taste-and-odor compounds in lakes and reservoirs and 

sample collection approaches for cyanobacterial toxins and 

taste-and-odor compounds. 

Acknowledgements

The authors thank Laura Simonson, USGS, and Ann St. 

Amand, of PhycoTech Inc., for their technical reviews, which 

provided valuable input and resulted in improved report content. We also thank Julie Hambrook Berkman for review and 

discussion of the initial report outline, Franceska Wilde for her 

ideas and technical comments throughout the development of 

this report, and Eric O’Brien for discussions on beach monitoring programs. Our appreciation also goes to Tim Miller, 

Chief of the USGS Office of Water Quality, for his support in 

this endeavor.

Selected References

American Water Works Association, 2004, Problem organisms 

in water: Identification and treatment (3d ed.): Denver, CO, 

American Water Works Association, 165 p.

Averett, R.C., and Schroder, L.J., 1994, A guide to the design 

of surface-water-quality studies: U.S. Geological Survey 

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Glossary 29

Glossary

A

Actinomycetes bacteria A group of aerobic, 

gram-positive bacteria that are largely terrestrial organisms associated with soils. The 

actinomycetes bacteria play a major role in 

the mineralization of organic matter in soils. 

Actinomycetes bacteria are not photosynthetic 

and are not part of the phytoplankton community in lakes and reservoirs.

Acute Single exposure to a relatively high 

dose of a toxic substance.

Algae Unicellular or simple multicellular photosynthetic organisms containing 

chlorophyll-a; there are numerous groups 

of algae, most of which are eukaryotic 

aquatic organisms. 

Allelopathy Inhibition or suppression of 

growth by one species of algae or plant by 

chemicals produced by another species. 

Aphotic zone Region where there is not 

enough light to support photosynthesis; 

extends from below the photic zone to the 

lake bottom.

B

Benthic Associated with the bottom of a 

lake or reservoir. After benthos, a term used to 

describe the bottom of a lake or reservoir.

Bioactive Indicates a substance has an effect 

on living tissue.

Biovolume The volume of an organism or 

group of organisms.

Bloom The term bloom, not specific to 

cyanobacteria, is inexact and subjective. Common definitions include: a large population or 

extremely high cell densities of phytoplankton 

(extremely high densities are typically defined 

as greater than 20,000 to 100,000 cells per 

milliliter); where proliferation of phytoplankton is dominated by a single or a few species; 

and visible accumulation of phytoplankton at 

the water surface.

Blue-green algae See cyanobacteria.

C

Chronic Repeated exposure to relatively low 

doses of a toxic substance over an extended 

period of time.

Cyanobacteria Cyanobacteria are true bacteria with a prokaryotic cell structure; however, 

cyanobacteria also have chlorophyll-a, a photopigment characteristic of eukaryotic algae 

and higher plants. Structurally the cyanobacteria are bacteria-like but functionally the 

cyanobacteria are algae-like. Cyanobacteria 

are typically sampled and analyzed as part 

of phytoplankton (algal) assemblages rather 

than bacterial assemblages in aquatic ecosystems. Cyanobacteria often are called bluegreen algae.

Cyanotoxins Any of a number of toxins produced by the cyanobacteria. The cyanotoxins 

include dermatoxins, hepatotoxins, and neurotoxins and impact a wide range of aquatic and 

terrestrial organisms, including humans. 

D

Dermatoxin A toxin that affects the skin and 

membrane tissues.

Diel Regular cycle over a 24-hour period.

E

Epilimnion The warm, buoyant upper layer 

of a stratified lake.

Eutrophic High nutrient content and levels 

of production.

Eutrophication Nutrient enrichment (particularly nitrogen and phosphorus) in aquatic 

ecosystems leading to increased productivity.

Extracellular Occurring outside of a cell.

F

G

H

Hepatotoxin A toxin that affects the liver.

Hypereutrophic Very high nutrient content 

and levels of production.

Hypolimnion The cold, dense bottom layer 

of a stratified lake; the hypolimnion often 

becomes anoxic (little or no dissolved oxygen) 

in productive systems. 
30 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

I

Icthyotoxin A toxin that affects fish or is 

specific to fish.

Intracellular Occurring within a cell.

Intraperitoneal Within the abdominal cavity.

J

K

Kemmerer Bottle A type of thief sampler 

used for collecting water samples at discrete 

depths. The sampler is held open at both ends 

and lowered vertically through the water column. At the desired depth a mechanism triggers the sampler and it closes, capturing the 

water at depth. The main difference between a 

Kemmerer Bottle and a Van Dorn Bottle is the 

mechanism used to open and close the bottle. 

See fig. 2-5A in chapter 2 (Lane and others, 

2003) of the U.S. Geological Survey National 

Field Manual (U.S. Geological Survey, 2006). 

L

M

Mesotrophic Moderate nutrient content and 

levels of production. 

Metalimnetic bloom An algal population 

that develops at the interface between the 

epilimnion and metalimnion in a stably stratified lake. Metalimnetic blooms most commonly occur in mesotrophic lakes where light 

penetrates into the metalimnion.

Metalimnion The middle layer of a stratified 

lake; the metalimnion is characterized by substantial decreases in temperature with depth.

Mixed depth The depth of turbulent mixing; 

may include all or only a portion of the water 

column depending on stratification, solar 

irradiance, and wind.

N

Neurotoxin A toxin that affects the central 

nervous system.

O

Oligotrophic Low nutrient content and levels 

of production.

P

Photic (euphotic) zone Region where there 

is enough light to support photosynthesis; 

extends from the surface to the depth were 

light is approximately one percent of that at 

the surface.

Phytoplankton Algae, including the 

cyanobacteria, suspended in the water column.

Q

R

S

Secondary metabolite Cellular products 

that are not directly involved in primary 

cellular processes that support growth and 

development.

Senesce (senescence) The process of 

growing old; aging.

Species A distinct kind of organism; the 

major division of genus.

Strain A group of organisms of the same 

species that have distinctive characteristics but 

are not considered a separate species.

T

Taste-and-odor compounds Compounds 

that produce objectionable tastes and odors in 

finished drinking water.

Thermocline The region where temperature 

change is greater than of equal to 1° C per 

meter; the terms thermocline and metalimnion 

are often used synonymously.

Trophic status Level of productivity in an 

ecosystem.

Turnover Complete isothermal mixing of a 

previously stratified lake. 

U

V

Van Dorn Bottle A type of thief sampler 

used for collecting water samples at discrete 

depths. The sampler is held open at both ends 

and lowered either vertically or horizontally 

through the water column. At the desired 

depth a mechanism triggers the sampler and it 

closes, capturing the water at depth. The main 

difference between a Van Dorn Bottle and 

a Kemmerer Bottle is the mechanism used 

to open and close the bottle. In addition, the 

Van Dorn Bottle may be held either vertically 

or horizontally, while the Kemmerer Bottle 

only can be used in the vertical position. See 

fig. 2-5B in chapter 2 (Lane and others, 2003) 

of the U.S. Geological Survey National Field 

Manual (U.S. Geological Survey, 2006). 

W

X

Y

Z
Appendixes
32 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Document the occurrence of cyanobacterial toxins and 

potential toxin producers in all primary recreational and 

drinking-water supply lakes and reservoirs within a state.

Design and Approach

Sample frequency: Samples will be collected monthly 

during June through September, the period when cyanobacterial populations typically peak.

Site location: In small lakes and reservoirs, samples will 

be collected at a single, representative open-water site near 

the deepest part of the lake or reservoir. In large lakes and 

reservoirs, samples will be collected at representative openwater sites in each of the main basins (lakes) or tributary arms, 

including a site near the dam (reservoirs). If present, samples 

from surface accumulations of cyanobacteria also will be collected.

Sample type: Integrated photic zone samples will be 

collected from open-water sites. Surface grab samples will be 

collected from surface accumulations.

Sampler used: Samples will be collected every meter 

throughout the photic zone using a vertical Van Dorn bottle. 

The samples from each depth will be composited in a churn. 

Appendix 1. Example Design and Approach for a Regional Reconnaissance 

Study 

Objective

An open-mouth bottle sampler will be used for surface grab 

samples.

Type of toxin analysis: Toxin samples will be analyzed for 

total (particulate plus dissolved) toxin concentrations.

Ancillary data collected: At each sample site, photographs and global positioning system (GPS) coordinates will 

be recorded and Secchi depth and vertical profiles of light 

(irradiance), water temperature, dissolved oxygen, conductivity, pH, turbidity, and in situ fluorescence will be measured. In 

addition to cyanobacterial toxins, each sample will be analyzed for phytoplankton community composition, abundance, 

and biovolume; chlorophyll; total phosphorus and nitrogen; 

and suspended sediment. 

Results: The results from this study can give an indication of how frequently cyanobacterial toxins occur in the 

state, which toxins are most common, and the range of typical 

concentrations. Potential cyanobacterial toxin producers also 

will be identified. General water-quality conditions in the 

state during peak periods of cyanobacterial abundance will be 

described. This study will not give a good indication of maximum toxin concentrations; monthly samples may miss periods 

of peak cyanobacterial toxin abundance. 
Appendix 1. Example Design and Approach for a Regional Reconnaissance Study 33

Field Form

Station Information

Station Name: Date: (MM/DD/YY)

_____/_____/_____

Time:

___ ___ ___ ___

Station Identification Number:

Project Name:

Sampled By: Samples Shipped By: Date: (MM/DD/YY)

_____/_____/_____

Sampling Information 

Sample Type: Regular Concurrent Replicate Sequential Replicate Split Replicate Field Blank

 Laboratory Blank Other:________________

Sample Collection: Integrated Photic Zone Sample Surface Accumulation Sample

Photic Depth:

Sample Depths:

Sampling Device: Vertical Van Dorn Open Mouth Bottle Sampler 

Sampling Method: Multiple Vertical Grab Sample

Physical Site Conditions

Lake Color: Brown Green Blue Clear Other_________________

Surface Accumulation Present: Yes No

Location:

Accumulation Sample Collected: Yes No

Sky: Clear Cloudy _____% Precipitation: Light Medium Heavy Rain Mist Fog

Wind: Calm Light Breeze Gusty Windy

Other Observations:

Related Sampling Activities

Variable Supporting Information Collected (Check)

Total Cyanobacterial Toxins Bottle Type: HDPE Amber Glass

Phytoplankton Preservative:

Preservative volume:__________

Chlorophyll Volume Filtered:

Nutrients

Suspended Sediment

Remarks:
34 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Field Form (continued)

Field Measurements

Latitude: __ __°__ __' __ __" N Secchi Depth: Air Temperature:

Longitude: __ __°__ __' __ __" W Surface Irradiance: Barometric Pressure:

Photic Depth: Other:

Vertical Profile 

Depth Irradiance

Water

Temperature

Dissolved 

Oxygen

Specific 

Conductance pH Turbidity

In situ

Fluorescence

Photographs/Observations:
Appendix 2. Example Design and Approach for a Recreational Monitoring Study 35

Monitor a recreational beach to determine if cyanobacterial toxins pose an acute exposure risk.

Design and Approach

Sample frequency: Samples will be collected weekly 

between Memorial Day and Labor Day, the periods of peak 

recreational use. If cyanobacteria accumulate in the beach 

area, daily samples will be collected until the accumulation 

dissipates. 

Site location: Samples will be collected from nine locations within the designated recreational area and composited. 

The nine locations will be determined by evenly dividing the 

recreational area into three transects that begin at the beach 

and extend into the water. Samples will be collected from three 

locations (ankle, knee, and chest deep) along each transect. 

Sample collection starts at the waters edge in ankle-deep water 

and continues into the water to approximately chest depth. 

Ankle-deep water samples will be collected approximately 

0.15 m below the surface. Knee- and chest-deep water samples 

will be collected approximately 0.30 m below the surface. If 

dense cyanobacterial accumulations are present outside of 

transect locations, an additional sample will be collected from 

the accumulation.

Appendix 2. Example Design and Approach for a Recreational Monitoring 

Study 

Objective

Sample type: Surface grab samples will be collected. 

At knee and chest deep locations samples will be collected 

approximately 15 centimeters below the water surface. Dense 

cyanobacterial accumulations will be sampled at the water 

surface.

Sampler used: Grab samples will be collected with an 

open-mouth bottle at each transect location and composited 

into a churn.

Type of toxin analysis: Toxin samples will be analyzed for 

total (particulate plus dissolved) toxin concentrations.

Ancillary data collected: At each sample transect global 

positioning system (GPS) coordinates will be recorded. In 

addition, photographs will be taken, and Secchi depth, water 

temperature, dissolved oxygen, conductivity, pH, turbidity, and 

in situ fluorescence will be measured at one central location in 

the recreational area.

Results: The results from this study will give an indication of when cyanobacterial toxin concentrations are elevated 

enough to cause human health concerns. Data collected may 

be used to post warnings or close recreational areas. Study 

results also will give an indication of the range of cyanotoxin 

concentrations in the recreational area and when peak values 

typically occur.
36 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Field Form 

Station Information

Station Name: Date: (MM/DD/YY)

_____/_____/_____

Time:

___ ___ ___ ___

Station Identification Number:

Project Name:

Sampled By: Samples Shipped By: Date: (MM/DD/YY)

_____/_____/_____

Sampling Information 

Sample Type: Regular Concurrent Replicate Sequential Replicate Split Replicate Field Blank

 Laboratory Blank Other:________________

Sample Collection: Surface Grab Sample Surface Accumulation Sample

Sampling Device: Open Mouth Bottle Sampler 

Sampling Method: Grab Sample

Physical Site Conditions

Lake Color: Brown Green Blue Clear Other_________________

Surface Accumulation Present: Yes No

Location:

Accumulation Sample Collected: Yes No

Sky: Clear Cloudy _____% Precipitation: Light Medium Heavy Rain Mist Fog

Wind: Calm Light Breeze Gusty Windy

Other Observations:

Field Measurements

Latitude: __ __°__ __' __ __" N Secchi Depth: pH:

Longitude: __ __°__ __' __ __" W Water Temperature: Turbidity:

Air Temperature: Dissolved Oxygen: In Situ Fluorescence:

Barometric Pressure: Specific Conductance: Other:

Photographs/Observations:

Sample Locations

Transect 1 Transect 2 Transect 3

Ankle Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Ankle Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Ankle Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Knee Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Knee Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Knee Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Chest Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Chest Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Chest Depth:

Latitude: __ __°__ __' __ __" N

Longitude: __ __°__ __' __ __" W

Remarks: Remarks: Remarks:

Related Sampling Activities

Variable Supporting Information Collected (Check)

Total Cyanobacterial Toxins Preservative:

Preservative volume:__________

Remarks:
Appendix 3. Example Design and Approach for an Interpretive Study 37

Appendix 3. Example Design and Approach for an Interpretive Study to 

Develop a Real-Time Model to Estimate Geosmin and 2-Methylisoborneol (MIB) 

Concentrations 

Objective

Describe the environmental factors that are associated 

with taste-and-odor episodes caused by cyanobacteria in a 

drinking-water supply reservoir and develop a real-time model 

to estimate the probability of geosmin and MIB concentrations 

exceeding the human detection threshold of 10 nanograms 

per liter.

Design and Approach

Sample frequency: Samples will be collected monthly 

November through April and bi-weekly May through October, 

when cyanobacteria typically are abundant in the drinkingwater supply reservoir. The sampling regime may need to be 

adjusted depending on potential ice cover. During a taste-andodor event, sampling frequency will be increased to weekly or 

semi-weekly. Sample collection will continue for 5 years.

Site location: The main study site will be located 

within the vicinity of the drinking water intake. Other locations also may be sampled if surface accumulations of 

cyanobacteria develop. 

Sample type: The type of samples collected will depend 

on vertical structure and distribution of cyanobacteria. Generally, when the water column is stratified, integrated samples 

will be collected from the epilimnion, metalimnion, and 

hypolimnion; when the water column is not stratified, integrated photic zone samples will be collected. Surface samples 

and discrete depth samples also may be collected depending 

on cyanobacterial distribution.

Sampler used: A pump will be used to collect depthintegrated samples and discrete depth samples. An openmouth bottle sampler will be used to collect surface samples. All sample types will be composited in a churn for 

sample processing.

Type of taste-and-odor analysis: Taste-and-odor samples 

will be analyzed for total and dissolved concentrations.

Ancillary data collected: Real-time water-quality monitors will be installed in the photic zone near the drinking water 

intake, and will measure lake level, light (irradiance), water 

temperature, dissolved oxygen, conductivity, pH, turbidity, and 

in situ fluorescence. An anemometer (wind meter) also will be 

installed at the site. Other meteorological data will be obtained 

daily from a local weather station and the timing of inflow 

events will be recorded. 

During each sampling, site photographs and global 

positioning system (GPS) coordinates will be recorded and 

Secchi depth and vertical profiles of light (irradiance), water 

temperature, dissolved oxygen, conductivity, pH, turbidity, and 

in situ fluorescence will be measured. In addition to taste-andodor compounds, each sample will be analyzed for total and 

dissolved cyanobacterial toxins, phytoplankton community 

composition, abundance, and biovolume, chlorophyll, actinomycetes bacteria, nutrients, suspended sediment, major ions, 

and alkalinity. 

Results: The results of this study will describe the seasonal patterns in the occurrence of taste-and-odor compounds 

and give an indication of when taste-and-odor episodes are 

most likely to occur. In addition, the relations between environmental variables, cyanobacterial community structure, and 

the occurrence of taste-and-odor compounds will be described, 

and may indicate potential management options. Real-time 

data and discrete water-quality samples will allow the development of a real-time model to estimate the probability that 

taste-and-odor compound concentrations will exceed the 

human detection threshold. This model can allow the drinkingwater treatment facility to adjust treatment accordingly and 

minimize the effects of taste-and-odor occurrences on drinking-water quality.
38 Guidelines for Design and Sampling for Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes and Reservoirs

Field Form

Station Information

Station Name: Date: (MM/DD/YY)

_____/_____/_____

Time:

___ ___ ___ ___

Station Identification Number:

Project Name:

Sampled By: Samples Shipped By: Date: (MM/DD/YY)

_____/_____/_____

Sampling Information 

Sample Type: Regular Concurrent Replicate Sequential Replicate Split Replicate Field Blank

 Laboratory Blank Other:________________

Sample Collection: Integrated Epilimnion Integrated Metalimnion Integrated Hypolimnion 

 Integrated Photic Zone Discrete Depth Surface Accumulation

Photic Depth:

Integrated Sample Depths:

Discrete Sample Depth:

Sampling Device: Pump Open Mouth Bottle Sampler 

Sampling Method: Multiple Vertical Point Sample Grab Sample

Physical Site Conditions

Lake Color: Brown Green Blue Clear Other_________________

Surface Accumulation Present: Yes No

Location:

Accumulation Sample Collected: Yes No

Sky: Clear Cloudy _____% Precipitation: Light Medium Heavy Rain Mist Fog

Wind: Calm Light Breeze Gusty Windy

Other Observations:

Related Sampling Activities

Variable Supporting Information Collected (Check)

Total Taste-and-Odor

Dissolved Taste-and-Odor

Total Cyanobacterial Toxins Bottle Type: HDPE Amber Glass

Dissolved Cyanobacterial Toxins Bottle Type: HDPE Amber Glass

Phytoplankton Preservative:

Preservative volume:__________

Chlorophyll Volume Filtered:

Actinomycetes Bacteria

Nutrients

Suspended Sediment

Major Ions

Alkalinity

Remarks:
Appendix 3. Example Design and Approach for an Interpretive Study 39

Field Form (continued)

Field Measurements

Latitude: __ __°__ __' __ __" N Secchi Depth: Air Temperature:

Longitude: __ __°__ __' __ __" W Surface Irradiance: Barometric Pressure:

Photic Depth: Other:

Vertical Profile 

Depth Irradiance

Water

Temperature

Dissolved 

Oxygen

Specific 

Conductance pH Turbidity

In situ

Fluorescence

Photographs/Observations:
Publishing support provided by:

Rolla Publishing Service Center

For more information concerning this publication, contact:

Director, USGS Kansas Water Science Center

4821 Quail Crest Place

Lawrence, KS 66049

(785) 842–9909

Or visit the Kansas Water Science Center Web site at:

http://ks.water.usgs.gov


Photo 1

Back cover. Photograph 1: Fish kill associated with a Microcystis bloom, August 8, 2006 (photograph taken by Jennifer L. Graham, 

U.S. Geological Survey). Photograph 2: Sampling a Planktothrix bloom, August 9, 2006 (photograph taken by Jennifer L. Graham, 

U.S. Geological Survey). Photograph 3: Wind-blown accumulation of Aphanizomenon and Microcystis in a recreational area, 

October 11, 2001 (photograph taken by Jennifer L. Graham, U.S. Geological Survey). 

Photo 2

Photo 3
Graham and others—Guidelines for Design and Sampling of Cyanobacterial Toxin and Taste-and-Odor Studies in Lakes—Scientific Investigations Report 2008–5038

Printed on recycled paper