Drone-based Water Sampling and Characterization of Three Freshwater Harmful Algal Blooms in the United States

Freshwater harmful algal blooms (HABs), caused mostly by toxic cyanobacteria, produce a range of cyanotoxins that threaten the health of humans and domestic animals. Climate conditions and anthropogenic influences such as agricultural run-off can alter the onset and intensity of HABs. Little is known about the distribution and spread of freshwater HABs. Current sampling protocols in some lakes involve teams of researchers that collect samples by hand from a boat and/or from the shoreline. Water samples can be collected from the surface, from discrete-depth collections, and/or from depth-integrated intervals. These collections are often restricted to certain months of the year, and generally are only performed at a limited number of collection sites. In lakes with active HABs, surface samples are generally sufficient for HAB water quality assessments. We used a unique DrOne Water Sampling SystEm (DOWSE) to collect water samples from the surface of three different HABs in Ohio (Grand Lake St Marys, GLSM and Lake Erie) and Virginia (Lake Anna), United States in 2019. The DOWSE consisted of a 3D-printed sampling device tethered to a drone (uncrewed aerial system, or UAS), and was used to collect surface water samples at different distances (10–100 m) from the shore or from an anchored boat. One hundred and eighty water samples (40 at GLSM, 20 at Lake Erie, and 120 at Lake Anna) were collected and analyzed from 18 drone flights. Our methods included testing for cyanotoxins, phycocyanin, and nutrients from surface water samples. Mean concentrations of microcystins (MCs) in drone water samples were 15.00, 1.92, and 0.02 ppb for GLSM, Lake Erie, and Lake Anna, respectively. Lake Anna had low levels of anatoxin in nearly all (111/120) of the drone water samples. Mean concentrations of phycocyanin in drone water samples were 687, 38, and 62 ppb for GLSM, Lake Erie, and Lake Anna, respectively. High levels of total phosphorus were observed in the drone water samples from GLSM (mean of 0.34 mg/L) and Lake Erie (mean of 0.12 mg/L). Lake Anna had the highest variability of total phosphorus with concentrations that ranged from 0.01 mg/L to 0.21 mg/L, with a mean of 0.06 mg/L. Nitrate levels varied greatly across sites, inverse with bloom biomass, ranging from below detection to 3.64 mg/L, with highest mean values in Lake Erie followed by GLSM and Lake Anna, respectively. Drones offer a rapid, targeted collection of water samples from virtually anywhere on a lake with an active HAB without the need for a boat which can disturb the surrounding water. Drones are, however, limited in their ability to operate during inclement weather such as rain and heavy winds. Collectively, our results highlight numerous opportunities for drone-based water sampling technologies to track, predict, and respond to HABs in the future

Drone-based water sampling and characterization of three freshwater harmful algal blooms in the United States

Freshwater harmful algal blooms (HABs), caused mostly by toxic cyanobacteria, produce a range of cyanotoxins that threaten the health of humans and domestic animals. Climate conditions and anthropogenic influences such as agricultural run-off can alter the onset and intensity of HABs. Little is known about the distribution and spread of freshwater HABs. Current sampling protocols in some lakes involve teams of researchers that collect samples by hand from a boat and/or from the shoreline. Water samples can be collected from the surface, from discrete-depth collections, and/or from depth-integrated intervals. These collections are often restricted to certain months of the year, and generally are only performed at a limited number of collection sites. In lakes with active HABs, surface samples are generally sufficient for HAB water quality assessments. We used a unique DrOne Water Sampling SystEm (DOWSE) to collect water samples from the surface of three different HABs in Ohio (Grand Lake St Marys, GLSM and Lake Erie) and Virginia (Lake Anna), United States in 2019. The DOWSE consisted of a 3D-printed sampling device tethered to a drone (uncrewed aerial system, or UAS), and was used to collect surface water samples at different distances (10–100 m) from the shore or from an anchored boat. One hundred and eighty water samples (40 at GLSM, 20 at Lake Erie, and 120 at Lake Anna) were collected and analyzed from 18 drone flights. Our methods included testing for cyanotoxins, phycocyanin, and nutrients from surface water samples. Mean concentrations of microcystins (MCs) in drone water samples were 15.00, 1.92, and 0.02 ppb for GLSM, Lake Erie, and Lake Anna, respectively. Lake Anna had low levels of anatoxin in nearly all (111/120) of the drone water samples. Mean concentrations of phycocyanin in drone water samples were 687, 38, and 62 ppb for GLSM, Lake Erie, and Lake Anna, respectively. High levels of total phosphorus were observed in the drone water samples from GLSM (mean of 0.34 mg/L) and Lake Erie (mean of 0.12 mg/L). Lake Anna had the highest variability of total phosphorus with concentrations that ranged from 0.01 mg/L to 0.21 mg/L, with a mean of 0.06 mg/L. Nitrate levels varied greatly across sites, inverse with bloom biomass, ranging from below detection to 3.64 mg/L, with highest mean values in Lake Erie followed by GLSM and Lake Anna, respectively. Drones offer a rapid, targeted collection of water samples from virtually anywhere on a lake with an active HAB without the need for a boat which can disturb the surrounding water. Drones are, however, limited in their ability to operate during inclement weather such as rain and heavy winds. Collectively, our results highlight numerous opportunities for drone-based water sampling technologies to track, predict, and respond to HABs in the future

Monitoring Cyanotoxin Production in the Western Basin of Lake Erie Using SPATT

The array of toxins and toxin concentrations produced by cyanobacteria in lakes may change on time scales that are too short to be adequately characterized by typical weekly water samples. We explored the use of Solid Phase Adsorption Toxin Tracking (SPATT) to provide a continuous profile of cyanobacterial toxin production in western Lake Erie. SPATT devices contain porous resin that passively adsorbs toxins over days or weeks potentially useful for detecting short-lived spikes in toxin levels, and unusual toxins and congeners. SPATT bags were deployed at two locations for durations of one and two weeks June-October, 2020. Grab samples for microcystin (MC) concentrations via ELISA and samples for qPCR analysis were collected weekly. SPATT bags were analyzed for 12 MC congeners, anatoxin-a (\u3c0.11 ppb), and cylindrospermopsin

Cyanobacteria and Cyanotoxins Occurrence and Removal from Five High-Risk Conventional Treatment Drinking Water Plants

An environmental protection agency EPA expert workshop prioritized three cyanotoxins, microcystins, anatoxin-a, and cylindrospermopsin (MAC), as being important in freshwaters of the United States. This study evaluated the prevalence of potentially toxin producing cyanobacteria cell numbers relative to the presence and quantity of the MAC toxins in the context of this framework. Total and potential toxin producing cyanobacteria cell counts were conducted on weekly raw and finished water samples from utilities located in five US states. An Enzyme-Linked Immunosorbant Assay (ELISA) was used to screen the raw and finished water samples for microcystins. High-pressure liquid chromatography with a photodiode array detector (HPLC/PDA) verified microcystin concentrations and quantified anatoxin-a and cylindrospermopsin concentrations. Four of the five utilities experienced cyanobacterial blooms in their raw water. Raw water samples from three utilities showed detectable levels of microcystins and a fourth utility had detectable levels of both microcystin and cylindrospermopsin. No utilities had detectable concentrations of anatoxin-a. These conventional plants effectively removed the cyanobacterial cells and all finished water samples showed MAC levels below the detection limit by ELISA and HPLC/PDA

Dhb Microcystins Discovered in USA Using an Online Concentration LC–MS/MS Platform

Based on current structural and statistical calculations, thousands of microcystins (MCs) can exist; yet, to date, only 246 MCs have been identified and only 12 commercial MC standards are available. Standard mass spectrometry workflows for known and unknown MCs need to be developed and validated for basic and applied harmful algal bloom research to advance. Our investigation focuses on samples taken in the spring of 2018 from an impoundment fed by Oser and Bischoff Reservoirs, Indiana, United States of America (USA). The dominant cyanobacterium found during sampling was Planktothrix agardhii. The goal of our study was to identify and quantify the MCs in the impoundment sample using chemical derivatization and mass spectrometry. Modifying these techniques to use online concentration liquid chromatography tandem mass spectrometry (LC–MS/MS), two untargeted MCs have been identified, [d-Asp3, Dhb7]-MC-LR and tentative [Dhb7]-MC-YR. [Dhb7]-MC-YR is not yet reported in the literature to date, and this was the first reported incidence of Dhb MCs in the United States. Furthermore, it was discovered that the commercially available [d-Asp3]-MC-RR standard was [d-Asp3, Dhb7]-MC-RR. This study highlights a workflow utilizing online concentration LC–MS/MS, high-resolution MS (HRMS), and chemical derivatization to identify isobaric MCs

Comparative Analysis of Microcystin Prevalence in Michigan Lakes by Online Concentration LC/MS/MS and ELISA

Fast and reliable workflows are needed to quantitate microcystins (MCs), a ubiquitous class of hepatotoxic cyanotoxins, so that the impact of human and environmental exposure is assessed quickly and minimized. Our goal was to develop a high-throughput online concentration liquid chromatography tandem mass spectrometry (LC/MS/MS) workflow to quantitate the 12 commercially available MCs and nodularin in surface and drinking waters. The method run time was 8.5 min with detection limits in the low ng/L range and minimum reporting levels between 5 and 10 ng/L. This workflow was benchmarked by determining the prevalence of MCs and comparing the Adda-ELISA quantitation to our new workflow from 122 samples representing 31 waterbodies throughout Michigan. The frequency of MC occurrence was MC-LA > LR > RR > D-Asp3-LR > YR > HilR > WR > D-Asp3-RR > HtyR > LY = LW = LF, while MC-RR had the highest concentrations. MCs were detected in 33 samples and 13 of these samples had more than 20% of their total MC concentration from MCs not present in US Environmental Protection Agency (US EPA) Method 544. Furthermore, seasonal deviations between the LC/MS/MS and Adda-ELISA data suggest Adda-ELISA cross-reacts with MC degradation products. This workflow provides less than 24-h turnaround for quantification and also identified key differences between LC/MS/MS and ELISA quantitation that should be investigated further

Identification and Quantification of Degradation Products of Microcystins using High-Resolution UHPLC-Orbitrap-MS

Sharmila I. Thenuwara1, Jyotshana Gautam2, Samuel Simpson1, Johnna Birbeck3, Judy A. Westrick3, Jason F. Huntley2, Dragan Isailovic1 1Department of Chemistry and Biochemistry, University of Toledo, Toledo, OH 43606 2Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, OH 43614 3Lumigen Instrumentation Center, Department of Chemistry, Wayne State University, Detroit, MI 48202 Microcystins (MCs) are heptapeptides produced by freshwater harmful algal blooms (HABs). Exposure to hepatotoxic MCs is a threat to humans and animals. Although conventional municipal water treatment processes can treat MC contaminated water, biodegradation of cyanotoxins using indigenous bacteria is cost-effective and environmentally friendly. We previously isolated and characterized five bacterial isolates from Lake Erie that degraded MC-LR into non-toxic fragments (Thees et al. 2018). Herein, degradation of MCs that are abundant in Lake Erie HABs, MC-LR and MC-RR, is investigated qualitatively and quantitatively using high-resolution LC-MS. UHPLC-Selected ion monitoring (SIM)-Orbitrap-MS analysis revealed two peaks at different retention times corresponding to m/z of a tetrapeptide degradation product. Fragmentation spectra of both peaks showed characteristic ADDA fragment with m/z 135.08. From the MS/MS spectrum, it was concluded that the peak with the shorter retention time than the substrate is linear tetrapeptide. These results indicate that the MC biodegradation mechanisms in Lake Erie bacteria may be distinct from those in other MC degrading bacteria. The enzymatic pathways and MC breakdown products are being investigated further