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USGS2008

Chapter A7. Section 7.5. Cyanobacteria in lakes and reservoirs: Toxin And taste-and-odor sampling guidelines

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. The purpose of NFM 7.5 is to provide guidelines for collecting, processing, and handling samples to be analyzed for cyanobacterial community composition (enumeration and identification) and total, particulate, and dissolved cyanobacterial toxins and taste-and-odor compounds in lakes and reservoirs (sections 7.5.5 through 7.5.9). Sections 7.5.1 through 7.5.4, however, are designed to provide some background information about cyanobacteria, including typical study designs and objectives related to the spatial and temporal occurrence of cyanobacteria (modified from Graham and others, 2008), in order to provide a useful context for sampling activities. The information presented here pertains to the occurrence of planktonic (free-floating) cyanobacteria in lakes and reservoirs. Each chapter of the National Field Manual is published separately and revised periodically. Newly published and revised chapters will be announced on the USGS Home Page on the World Wide Web under 'New Publications of the U.S. Geological Survey.'

Techniques of Water-Resources Investigations↗

USGS1980

The susceptibility of the mallard duck (Anas platyrhynchos) to Clostridium botulinum C2 toxin

Most strains of Clostridium botulinum type C, after having lost their capacity to produce their dominant toxin (C 1 ) as a result of being“cured”of their prophages, continue to produce C 2 , a trypsin-activable toxin reported by other investigators. While of relatively low toxicity when administered perorally to the adult mallard duck ( Anas platyrhynchos ), it was highly toxic when given parenterally. By the intravenous route, for example, it was more than 1, 000 times as toxic as C 1 toxin by the same route, when compared on the basis of mouse intraperitoneal toxicity. The cause of death in every instance was massive pulmonary edema and hemorrhage rather than the respiratory paralysis that occurs in C 1 intoxication. 抄録

Japanese Journal of Medical Science and Biology↗

USGS2022

Paralytic shellfish toxins associated with Arctic Tern mortalities in Alaska

Harmful algal blooms produce biotoxins that can injure or kill fish, wildlife, and humans. These blooms occur naturally but have intensified in many locations globally due to recent climatic changes, including ocean warming. Such changes are especially pronounced in northern regions, where the effects of paralytic shellfish toxins (PSTs) on marine wildlife are of growing concern. In Alaska, seabird mortality events have increased in frequency, magnitude, and duration since 2015 alongside anomalously high ocean temperatures. Although starvation has been implicated as the apparent cause of death in many of these die-offs, saxitoxin (STX) and other PSTs have been identified as possible contributing factors. Here, we describe a mortality event at a nesting colony of Arctic Terns ( Sterna paradisaea ) near Juneau, Alaska in 2019 and report elevated concentrations of PSTs in bird, forage fish, and mussel samples. Concentrations of STX and other PSTs in tern tissues (2.5–51.2 µg 100g −1 STX-equivalents [STX-eq]) were of similar magnitude to those reported from other PST-induced bird die-offs. We documented high PST concentrations in blue mussels (>11,000 µg 100g −1 STX-eq; Mytilus edulis spp.) collected from nearby beaches, as well as in forage fish (up to 494 µg 100g −1 STX-eq) retrieved from Arctic Tern nests, thereby providing direct evidence of PST exposure via the terns’ prey. At maximum concentrations measured in this study, a single 5 g Pacific Sand Lance ( Ammodytes personatus ) could exceed the median lethal STX dose (LD 50 ) currently estimated for birds, offering strong support for PSTs as a likely source of tern mortality. In addition to describing this localized bird mortality event, we used existing energetics data from adult and nestling Arctic Terns to calculate estimated cumulative daily PST exposure based on ecologically relevant concentrations in forage fish. Our estimates revealed potentially lethal levels of PST exposure even at relatively low (≤30 ug 100g −1 STX-eq) toxin concentrations in prey. These findings suggest that PSTs present a significant hazard to Arctic Terns and other northern seabirds and should be included in future investigations of avian mortality events as well as assessments of population health.

Alaska↗

USGS2025

Climate-driven deoxygenation of lakes alters the nutrient-toxin profile of a food fish

Climate change is rapidly altering fisheries supporting aquatic ecosystems. The implications for food security depend not only on harvest biomass but also concentrations of nutrients and toxins in fish. Using brook trout from Adirondack lakes (New York, USA), we tested whether ongoing lake deoxygenation trends will affect fish muscle omega-3 fatty acids, selenium, and mercury concentrations. Across space (16 lakes: 1 year) and time (6 years: 1 lake), anoxia decreased selenium and was associated with elevated fish mercury, with no effect on omega-3 content. Because selenium may mitigate some end points of mercury toxicity, highly variable Se:Hg molar ratios (0.70–35.79) in neighboring lakes may have health risk implications. For fish consumers, ongoing lake deoxygenation under climate change could potentially reduce selenium intake while enhancing mercury exposure. Simultaneous alteration of beneficial compounds and toxins by environmental change complicates the development of fish consumption advisories to safeguard public health in a warming world.

New York↗

USGS2013

Association of toxin-producing Clostridium botulinum with the macroalga Cladophora in the Great Lakes

Avian botulism, a paralytic disease of birds, often occurs on a yearly cycle and is increasingly becoming more common in the Great Lakes. Outbreaks are caused by bird ingestion of neurotoxins produced by Clostridium botulinum, a spore-forming, gram-positive, anaerobe. The nuisance, macrophytic, green alga Cladophora (Chlorophyta; mostly Cladophora glomerata L.) is a potential habitat for the growth of C. botulinum. A high incidence of botulism in shoreline birds at Sleeping Bear Dunes National Lakeshore (SLBE) in Lake Michigan coincides with increasingly massive accumulations of Cladophora in nearshore waters. In this study, free-floating algal mats were collected from SLBE and other shorelines of the Great Lakes between June and October 2011. The abundance of C. botulinum in algal mats was quantified and the type of botulism neurotoxin (bont) genes associated with this organism were determined by using most-probable-number PCR (MPN-PCR) and five distinct bont gene-specific primers (A, B, C, E, and F). The MPN-PCR results showed that 16 of 22 (73%) algal mats from the SLBE and 23 of 31(74%) algal mats from other shorelines of the Great Lakes contained the bont type E (bont/E) gene. C. botulinum was present up to 15 000 MPN per gram dried algae based on gene copies of bont/E. In addition, genes for bont/A and bont/B, which are commonly associated with human diseases, were detected in a few algal samples. Moreover, C. botulinum was present as vegetative cells rather than as dormant spores in Cladophora mats. Mouse toxin assays done using supernatants from enrichment of Cladophora containing high densities of C. botulinum (>1000 MPN/g dried algae) showed that Cladophora-borne C. botulinum were toxin-producing species (BoNT/E). Our results indicate that Cladophora provides a habitat for C. botulinum, warranting additional studies to better understand the relationship between this bacterium and the alga, and how this interaction potentially contributes to botulism outbreaks in birds.

Great Lakes↗

USGS1999

In situ detection of the Clostridium botulinum type C1 toxin gene in wetland sediments with a nested PCR assay

A nested PCR was developed for detection of the Clostridium botulinum type C 1 toxin gene in sediments collected from wetlands where avian botulism outbreaks had or had not occurred. The C 1 toxin gene was detected in 16 of 18 sites, demonstrating both the ubiquitous distribution of C. botulinum type C in wetland sediments and the sensitivity of the detection assay.

Applied and Environmental Microbiology↗

USGS2018

Water-quality conditions with an emphasis on cyanobacteria and associated toxins and taste-and-odor compounds in the Kansas River, Kansas, July 2012 through September 2016

Cyanobacteria cause a multitude of water-quality concerns, including the potential to produce toxins and taste-and-odor compounds that may cause substantial economic and public health concerns, and are of particular interest in lakes, reservoirs, and rivers that are used for drinking-water supply. Extensive cyanobacterial blooms typically do not develop in the Kansas River; however, reservoirs in the lower Kansas River Basin occasionally develop blooms that may affect downstream water quality. During July 2012 through September 2016, continuous and (or) discrete water-quality data were collected at several sites (Wamego, De Soto, and three main reservoir-fed tributaries) on the Kansas River to characterize the sources, frequency and magnitude of occurrence, and causes of cyanobacteria, cyanobacterial toxins, and taste-and-odor compounds and to develop a real-time notification system of changing water-quality conditions that may affect drinking-water treatment. Algal biomass, as estimated by chlorophyll, was consistently higher at the downstream De Soto site than the upstream Wamego site. Higher algal biomass at the De Soto site likely was caused by algal growth during downstream transport without major losses due to grazing by aquatic organisms or other processes. Algal biomass at the Wamego and De Soto sites was negatively correlated with streamflow and total and bioavailable nutrient concentrations. The negative association between algal biomass and nutrients in the Kansas River likely reflects the relatively strong positive association between nutrient concentrations and streamflows. Cyanobacteria were relatively common in the Kansas River but rarely dominated the algal community. Like overall algal biomass, cyanobacterial abundances typically were higher at the De Soto site than the Wamego site. Cyanobacterial abundances generally peaked in late summer or early fall (July through October), with smaller peaks occasionally observed in spring (April through May). Cyanobacteria in the Kansas River rarely exceeded 20,000 cells per milliliter, the abundance at which cyanobacteria may become a concern for drinking-water treatment. Relations between cyanobacterial abundance and streamflow, turbidity, and nutrients in the Kansas River were similar to those between chlorophyll and total phytoplankton abundance, indicating the same processes that influence overall algal biomass and dynamics also are influencing cyanobacteria. The cyanotoxin microcystin was detected in about 27 percent of the samples collected from Kansas River tributary and main-stem sites. Cylindrospermopsin was detected in one sample from the De Soto site. Microcystin occurrence and concentration were similar between the Wamego and De Soto sites. Concentrations exceeded the U.S. Environmental Protection Agency health advisory guidance values for finished drinking water of 0.3 (for bottle-fed infants and pre-school children) and 1.6 micrograms per liter (μg/L; for school-age children and adults) in 6 percent or less of samples collected. These guidance values are for finished drinking water and are not directly applicable to observed environmental concentrations but do provide a benchmark for comparison. Microcystin was detected most often and had the highest concentrations during summer. Though seasonal patterns in microcystin occurrence were generally consistent, seasonal maxima varied by an order of magnitude across years. The taste-and-odor compounds geosmin and 2-methylisoborneol (MIB) were detected in about 78 and 43 percent of samples, respectively, collected across all sites (main stem and tributaries). Geosmin and MIB occurrence and concentration varied considerably between the Wamego and De Soto sites. Geosmin was detected in about 67 percent of Wamego samples and 81 percent of De Soto samples. The human detection threshold of 5 nanograms per liter (ng/L) was exceeded for geosmin in about 11 and 17 percent of the samples collected at the Wamego and De Soto sites, respectively. Geosmin was detected during all months of the year at both sites, and there were no clear seasonal patterns. MIB was detected less frequently in the Kansas River than geosmin and was observed in about 42 percent of Wamego samples and 33 percent of De Soto samples. Concentrations exceeded 5 ng/L in about 7 and 5 percent of samples from the Wamego and De Soto sites, respectively. As observed for geosmin, there were no clear seasonal patterns in MIB occurrence or concentration. There seems to be a connection between microcystin detections in the Kansas River and occurrence of microcystin in upstream reservoirs (and tributary streams). Microcystin concentrations greater than 0.3 μg/L may be likely during the summer when streamflow is less than 3,000 cubic feet per second (ft 3 /s) and contributions from Milford Lake exceed about 30 percent of total flow in the Kansas River. Observed microcystin concentrations typically were higher at the De Soto site than the Wamego or tributary sites during 2012 through 2016, indicating cyanobacteria may continue to grow and produce microcystin once introduced to the Kansas River. The spatial and temporal patterns in geosmin and MIB occurrence and concentration were more complex than microcystin. There were no clear connections between geosmin and MIB occurrence in the Kansas River and potential upstream reservoir (or tributary stream) sources. Likewise, there was not a clear relation between algal biomass, cyanobacteria, or actinomycetes bacteria and taste-and-odor events in the Kansas River. Geosmin and MIB were not strongly correlated with any measured environmental variable at either Kansas River site. Continuous water-quality data may be used independently or in combination with regression models to provide information on changing water-quality conditions that may affect drinking-water treatment processes or recreational activities on the Kansas River. For example, logistic regression model outputs and continuous water-quality data may both be indicative of the potential for microcystin events. Logistic regression models that are estimating a high probability of microcystin occurrence at concentrations above 0.1 μg/L can be used as one indicator. Streamflows less than 3,000 ft 3 /s during upstream reservoir releases during periods with low turbidity and increased chlorophyll fluorescence, specific conductance, and pH values may also be indicative of microcystin events. Advanced or near-real-time notification may inform proactive, rather than reactive, management strategies when water-quality conditions are changing rapidly or are likely to cause cyanobacteria-related events.

Kansas↗

USGS2021

Investigation of algal toxins in a multispecies seabird die-off in the Bering and Chukchi seas

Between 2014 and 2017, widespread seabird mortality events were documented annually in the Bering and Chukchi seas, concurrent with dramatic reductions of sea ice, warmer than average ocean temperatures, and rapid shifts in marine ecosystems. Among other changes in the marine environment, harmful algal blooms (HABs) that produce the neurotoxins saxitoxin (STX) and domoic acid (DA) have been identified as a growing concern in this region. Although STX and DA have been documented in Alaska (US) for decades, current projections suggest that the incidence of HABs is likely to increase with climate warming and may pose a threat to marine birds and other wildlife. In 2017, a multispecies die-off consisting of primarily Northern Fulmars ( Fulmarus glacialis ) and Short-tailed Shearwaters ( Ardenna tenuirostris ) occurred in the Bering and Chukchi seas. To evaluate whether algal toxins may have contributed to bird mortality, we tested carcasses collected from multiple locations in western and northern Alaska for STX and DA. We did not detect DA in any samples, but STX was present in 60% of all individuals tested and in 88% of Northern Fulmars. Toxin concentrations in Northern Fulmars were within the range of those reported from other STX-induced bird die-offs, suggesting that STX may have contributed to mortalities. However, direct neurotoxic action by STX could not be confirmed and starvation appeared to be the proximate cause of death among birds examined in this study.

Journal of Wildlife Diseases↗

USGS2026

Tracking toxins: A pilot investigation of cyanotoxins in north-central Tennessee’s surface waters and wells

Cyanobacterial toxins (cyanotoxins) threaten aquatic ecosystems and human health, yet the factors influencing their production and distribution in freshwater remain unclear. In north-central Tennessee, nutrient-rich runoff from agricultural and urban areas, combined with a karst landscape that supports drinking and recreational water use, heightens the need to understand cyanotoxin behavior. To examine cyanotoxin patterns, the U.S. Geological Survey and the Tennessee Department of Environment and Conservation monitored 18 sites, including two wells under the influence of surface water, every two weeks from September 2022 to November 2024. At least one cyanotoxin was detected at all sites, with the highest concentrations in deep reservoirs and lower levels in shallow systems. Most detections occurred during summer and fall, aligning with high temperatures and rapid-onset drought. Statistical analysis indicated that increased specific conductivity and pH raised the likelihood of detecting total microcystin, likely resulting from drought conditions and nutrient-laden runoff. Additionally, dissolved microcystin showed an inverse relationship with Cumberland River water levels, and principal component analysis showed that Secchi depth, chlorophyll a, pH, temperature, and conductivity explained most water quality variability. These results help increase understanding of cyanotoxin distribution and associated water quality conditions during detections to guide future freshwater cyanotoxin monitoring studies.

Tennessee↗

USGS2012

Plant toxins and trophic cascades alter fire regime and succession on a boral forest landscape

Two models were integrated in order to study the effect of plant toxicity and a trophic cascade on forest succession and fire patterns across a boreal landscape in central Alaska. One of the models, ALFRESCO, is a cellular automata model that stochastically simulates transitions from spruce dominated 1 km2 spatial cells to deciduous woody vegetation based on stochastic fires, and from deciduous woody vegetation to spruce based on age of the cell with some stochastic variation. The other model, the ‘toxin-dependent functional response’ model (TDFRM) simulates woody vegetation types with different levels of toxicity, an herbivore browser (moose) that can forage selectively on these types, and a carnivore (wolf) that preys on the herbivore. Here we replace the simple succession rules in each ALFRESCO cell by plant–herbivore–carnivore dynamics from TDFRM. The central hypothesis tested in the integrated model is that the herbivore, by feeding selectively on low-toxicity deciduous woody vegetation, speeds succession towards high-toxicity evergreens, like spruce. Wolves, by keeping moose populations down, can help slow the succession. Our results confirmed this hypothesis for the model calibrated to the Tanana floodplain of Alaska. We used the model to estimate the effects of different levels of wolf control. Simulations indicated that management reductions in wolf densities could reduce the mean time to transition from deciduous to spruce by more than 15 years, thereby increasing landscape flammability. The integrated model can be useful in estimating ecosystem impacts of wolf control and moose harvesting in central Alaska.

Alaska↗

USGS2021

Cyanotoxin mixture models: Relating environmental variables and toxin co-occurrence to human exposure risk

Toxic cyanobacterial blooms, often containing multiple toxins, are a serious public health issue. However, there are no known models that predict a cyanotoxin mixture (anatoxin-a, microcystin, saxitoxin). This paper presents two cyanotoxin mixture models (MIX) and compares them to two microcystin (MC) models from data collected in 2016–2017 from three recurring cyanobacterial bloom locations in Kabetogama Lake, Voyageurs National Park (Minnesota, USA). Models include those using near-real-time environmental variables (readily available) and those using additional comprehensive variables (based on laboratory analyses). Comprehensive models (R 2 = 0.87 MC; R 2 = 0.86 MIX) explained more variability than the environmental models (R 2 = 0.58 MC; R 2 = 0.57 MIX). Although neither MIX model was a better fit than the MC models, the MIX models produced no false negatives in the calibration dataset, indicating that all observations above regulatory guidelines were simulated by the MIX models. This is the first known use of Virtual Beach software for a cyanotoxin mixture model, and the methods used in this paper may be applicable to other lakes or beaches.

Minnesota↗

USGS2015

Prevalence of toxin-producing Clostridium botulinum associated with the macroalga Cladophora in three Great Lakes: Growth and management

The reemergence of avian botulism caused by Clostridium botulinum type E has been observed across the Great Lakes in recent years. Evidence suggests an association between the nuisance algae, Cladophora spp., and C. botulinum in nearshore areas of the Great Lakes. However, the nature of the association between Cladophora and C. botulinum is not fully understood due, in part, to the complex food web interactions in this disease etiology. In this study, we extensively evaluated their association by quantitatively examining population size and serotypes of C. botulinum in algal mats collected from wide geographic areas in lakes Michigan, Ontario, and Erie in 2011–2012 and comparing them with frequencies in other matrices such as sand and water. A high prevalence (96%) of C. botulinum type E was observed in Cladophora mats collected from shorelines of the Great Lakes in 2012. Among the algae samples containing detectable C. botulinum , the population size of C. Botulinum type E was 10 0 –10 4 MPN/g dried algae, which was much greater (up to 10 3 fold) than that found in sand or the water column, indicating that Cladophora mats are sources of this pathogen. Mouse toxinantitoxin bioassays confirmed that the putative C. botulinum belonged to the type E serotype. Steam treatment was effective in reducing or eliminating C. botulinum type E viable cells in Cladophora mats, thereby breaking the potential transmission route of toxin up to the food chain. Consequently, our data suggest that steam treatment incorporated with a beach cleaning machine may be an effective treatment of Cladophora -borne C. botulinum and may reduce bird mortality and human health risks.

Lake Erie, Lake Michigan, Lake Ontario↗

USGS2009

Gene and antigen markers of Shiga-toxin producing E. coli from Michigan and Indiana river water: Occurrence and relation to recreational water quality criteria

The relation of bacterial pathogen occurrence to fecal indicator bacteria (FIB) concentrations used for recreational water quality criteria (RWQC) is poorly understood. This study determined the occurrence of Shiga-toxin producing Escherichia coli (STEC) markers and their relation to FIB concentrations in Michigan and Indiana river water. Using 67 fecal coliform (FC) bacteria cultures from 41 river sites in multiple watersheds, we evaluated the occurrence of five STEC markers: the Escherichia coli (EC) O157 antigen and gene, and the STEC virulence genes eae A, stx 1, and stx 2. Simple isolations from selected FC cultures yielded viable EC O157. By both antigen and gene assays, EC O157 was detected in a greater proportion of samples exceeding rather than meeting FC RWQC ( P < 0.05), but was unrelated to EC and enterococci RWQC. The occurrence of all other STEC markers was unrelated to any FIB RWQC. The eaeA , stx2, and stx1 genes were found in 93.3, 13.3, and in 46.7% of samples meeting FC RWQC and in 91.7, 0.0, and 37.5% of samples meeting the EC RWQC. Although not statistically significant, the percentage of samples positive for each STEC marker except stx1 was lower in samples that met, as opposed to exceeded, FIB RWQC. Viable STEC were common members of the FC communities in river water throughout southern Michigan and northern Indiana, regardless of FIB RWQC. Our study indicates that further information on the occurrence of pathogens in recreational waters, and research on alternative indicators of their occurrence, may help inform water-resource management and public health decision-making.

Journal of Environmental Quality↗

USGS2019

Real-time assessments of water quality—A nowcast for Escherichia coli and cyanobacterial toxins

Threats to our recreational and drinking waters include disease-causing (pathogenic) organisms from fecal contamination and toxins produced by some species of cyanobacteria (cyanotoxins) that can cause acute and (or) chronic illnesses. Because traditional laboratory methods for detecting these threats take too long for prompt public health protection, tools for real-time assessments are needed to protect public health. To address this need, the U.S. Geological Survey is collaborating with State and local partners to develop models that provide real-time estimates of Escherichia coli ( E . coli ) (for pathogens) and (or) microcystin (for freshwater cyanotoxins) levels at inland and Great Lakes beaches and drinking-water intakes. Model results are then used to inform the public of water-quality conditions in near-real time through the Great Lakes NowCast ( https://ny.water.usgs.gov/maps/nowcast/ ). Behind the scenes, the NowCast provides speed and efficiency for managers by automating data management and standardizing methods among agencies.

Fact Sheet↗

USGS2022

Detecting algal toxins and organic contaminants of concern in the environment

The U.S. Geological Survey (USGS) Kansas Water Science Center Organic Geochemistry Research Laboratory (OGRL) was established in 1987. The OGRL is a multidisciplinary program that contributes knowledge about the distribution, fate, transport, and effects of new and understudied organic compounds that may affect human health and (or) ecosystems. The OGRL consists of two units: Algal and Other Environmental Toxins Unit and Environmental Organic Chemistry Unit. The OGRL does independent and collaborative research, develops robust analytical methods, and provides fee-for-service analytical laboratory analyses.

Fact Sheet↗

USGS2015

Infectious diseases, parasites, and biological toxins in sea ducks

This chapter addresses disease agents in the broad sense, including viruses, bacteria, fungi, protozoan and helminth parasites, and biological toxins. Some of these agents are known to cause mortality in sea ducks, some are thought to be incidental findings, and the significance of others is yet poorly understood. Although the focus of the chapter is on free-living sea ducks, the study of disease in this taxonomic group has been relatively limited and examples from captive sea ducks and other wild waterfowl are used to illustrate the pathogenicity of certain diseases. Much of the early work in sea ducks consisted of anecdotal and descriptive reports of parasites, but it was soon recognized that diseases such as avian cholera, renal coccidiosis, and intestinal infections with acanthocephalans were causes of mortality in wild populations. More recently, adenoviruses, reoviruses, and the newly emergent Wellfleet Bay virus, for example, also have been linked to die-offs of sea ducks. Declining populations of animals are particularly vulnerable to the threats posed by disease and it is important that we improve our understanding of the significance of disease in sea ducks. To conclude, we offer our recommendations for future directions in this field.

Book chapter↗

USGS1938

The distribution of botulinus toxin in duck sickness areas

One of the most important events in the history of the diseases of wildfowl wa the discovery of the cause of western duck sickness. Determination that this malady of migratory birds is not a chemical poisoning, as had been previously suspected, was made by Kalmbach and others in 1930-31. Thtat it is a food poisoning resulting from toxins produced by botulinus organisms was proved beyond question. An extensive review of the work leading up to this determination cannot be made in this paper, but some orientation is necessary to a proper appreciation of the subject.

Conference Paper↗

USGS2025

Diel and spatial variability in cyanobacterial composition, gene abundance, and toxin concentration: A pilot study

We designed a pilot field study to assess relations between sunlight, cyanobacteria, and cyanotoxins. In 2021, we collected day (07:00 h, 10:00 h, 13:00 h, 16:00 h) and night samples (19:00 h, 22:00 h, 01:00 h, 04:00 h) at two locations in Kabetogama Lake, MN, USA. One sample set was collected from the lakeward end of a boat dock and the other on the nearby shoreline. Cyanobacterial phylogenetic eDNA differences over 24 h (pseudo F = 2.0938, p = 0.127) were not significant. Copies of anatoxin ( anaC ) and microcystin ( mcyE ) synthetase genes varied significantly over the sampling times at the dock (Friedman Χ 2 = 15.01, df = 7, p = 0.036; Friedman Χ 2 = 19.22, df = 7, p = 0.008) and the shoreline (Friedman Χ 2 = 19.33, df = 7, p = 0.007; Friedman Χ 2 = 20.56, df = 7, p = 0.005), with the highest anaC counts occurring during the night for both sites. Additionally, the highest total and dissolved microcystin concentrations occurred at night. Despite the proximity of the sampling locations, cyanobacterial phylogenetic eDNA results indicate that the variability between sites (pseudo-F = 27.547, p = 0.001) were greater than temporal differences over 24 h (pseudo F = 2.0938, p = 0.127). Understanding the effect of diel and spatial variability may help researchers and resource managers make informed decisions about sampling and potential exposure.

Minnesota↗