St. Louis River Area of Concern Remedial Action Plan Project 7.02

This final report details the results of the Wisconsin Department of Natural Resources (WDNR) Project Bacterial Source Tracking at Impaired Beaches in the St. Louis River Area of Concern, hereafter referred to as the Bacterial Source Tracking Project. This project is described in the St. Louis River Area of Concern (AOC) Remedial Action Plan (RAP) as Project 7.02, the goal of which was to conduct bacterial source identification via DNA analysis to determine if bacterial contaminants were of human origin, and if necessary, recommend additional restoration options. The field sampling portion of this project took place on six impaired beaches in the St. Louis River AOC, on the State of Wisconsin or State of Minnesota 2014 and 2016 303(d) lists of non-attaining waters due to contamination by pathogens. Primary environmental data generated during this project includes Sanitary Survey (SS) data, concentration of Escherichia coli in surface water samples, and origin of measured E. coli via quantitative polymerase chain reaction (qPCR) analysis. The SS and E. coli concentration data were generated by the Lake Superior Research Institute (LSRI,University of Wisconsin-Superior), while the DNA marker data were generated by the by Dr. Sandra McLellan’s laboratory at the University of Wisconsin-Milwaukee’s School of Freshwater Sciences. The data generated from this project will be used by the St. Louis River AOC Coordinators and Beneficial Use Impairment (BUI) 7 Technical Team to determine if BUI 7 removal targets have been met.Water Quality Bureau; Wisconsin Department of Natural Resources; 1701 N. 4th Street; Superior, WI 5488

Identification of naturally-occurring fluoride and selected metals in northwest Wisconsin groundwater

Northwest Wisconsin, encompassing the 11 counties of Ashland, Barron, Bayfield, Burnett, Douglas, Iron, Polk, Price, Rusk, Sawyer, and Washburn, has a large proportion of the population who obtain their drinking water from groundwater. There are over 56,000 public and private wells within this region of the state, however, the region is under-studied in terms of groundwater quality relative to other areas in Wisconsin. Fluoride and the metals iron, manganese, aluminum, arsenic, and lead, if present at high enough concentrations in groundwater, could impact public health. Little is known regarding baseline concentrations of these potential contaminants in northwestern Wisconsin aquifers. The objectives of this study, known as the Northwest Wisconsin Groundwater Monitoring Project, were to monitor naturally-occurring fluoride concentrations in groundwater samples collected from 11 counties in northwestern Wisconsin, determine concentrations of iron, manganese, aluminum, arsenic, and lead in a subset of these samples, foster public awareness among residents of the region about the need for regular private well testing, and increase the publicly-available groundwater data within the region. This monitoring study used a random sample design, and targeted 704 fluoride samples and 115 metals samples total. All of the samples were collected by volunteers who were recruited to participate in the project through a combination of social media postings, press releases, and outreach events. Sample kits containing sample bottles, instructions for collection, sample collection form, and a pre-paid shipping label were assembled and delivered to each of 26 distribution sites located throughout the 11-county study area. The data from 450 of 704 fluoride and 80 of 115 metals samples were reported. Samples were collected from all 11 counties in the study area, and were largely collected from drilled wells constructed 26 – 50 years ago, with a depth of 51 – 100 feet. None of the samples collected and analyzed for fluoride exceeded the Wisconsin Groundwater Quality Enforcement Standard (ES) of 4.0 mg/L and less than 1% exceeded the Preventative Action Limit (PAL) of 0.8 mg/L (Wis. Adm. Code Ch. NR 140). Iron concentrations were high in a number of samples, with 20% of samples collected exceeding the 0.3 mg/L ES. In 5% of samples collected, the ES for manganese, 300 µg/L, was exceeded. Overall, aluminum concentrations were low throughout the study area, but one sample exceeded the ES of 200 µg/L. The maximum arsenic concentration measured in this study was 10.0 µg/L, which is the state ES. However, 38.8% of samples had an arsenic concentration at or above the PAL (1 µg/L). There was a single sample that exceeded the ES for lead (15 µg/L) and 11.3% of samples had a lead concentration at or above the 1.5 µg/L PAL. To the authors’ knowledge, this study represents the first baseline groundwater monitoring effort encompassing the northwest Wisconsin region. With the exception of fluoride and arsenic, the ES was exceeded for all parameters measured in this study in at least one sample collected within the study area. This study utilized a random sample design, and the next logical step is to conduct targeted sampling of areas with contaminant-level metals concentrations in groundwater samples.Funding for this work was provided by the Wisconsin Department of Natural Resources via the Wisconsin Groundwater Coordinating Council

Tests of the LED Light Activated Titanium Dioxide Bench-Scale Ballast Water Treatment Process

This technical report presents findings from bench-scale tests evaluating the performance of the LED Light Activated Titanium Dioxide Technology, hereafter LED TiO2, developed by YJB LED Professional Services of Crosslake, Minnesota, USA. Researchers conducted the bench-scale evaluation beginning in July 2019 and ending in September 2019 at the Lake Superior Research Institute (LSRI) of the University of Wisconsin-Superior (UWS) in Superior, Wisconsin, USA. The LED TiO2 treatment process applies light emitting diodes (LED) to activate a photocatalytic coating that creates a bacteriostatic, fungistatic, and algastatic environment. Biological effectiveness testing was completed with the algae, Selenastrum capricornutum and pathogen indicator organisms, Escherichia coli and Enterococcus faecium in lab water. The system was found to be effective at treating microbes in highly-transparent/low-suspended solids water, but was less effective at treating algae

Kria Ionizer Test Report

This technical report presents findings from bench-scale tests evaluating the performance of the Kria Ionizer Superoxide Generator Model DG-100, hereafter Kria Ionizer, developed by EcoUSA, LLC of Denver, Colorado, United States of America. This evaluation was the first to assess the Kria Ionizer as a potential, in-tank, recirculating ballast water treatment method for the Laurentian Great Lakes. The evaluation began in May 2019 and ended in September 2019 at the Lake Superior Research Institute (LSRI) of the University of Wisconsin-Superior (UWS) in Superior, Wisconsin, USA. The treatment technology uses atmospheric oxygen to create superoxide (O2-). According to the developer, the injection of this superoxide into water not only creates a supersaturated oxygenated environment but can also promote cavitation and microbubble reactions upon discharge. The system can be used for oxygenation of fish farms and for remediation of water bodies containing harmful cyanobacteria, wastewater bacteria, and organic contaminants. Previous testing has been completed in the lab and field setting for these previously mentioned applications, but the testing described herein is the first testing evaluating Kria Ionizer’s potential application for ballast water treatment. The ability of the Kria Ionizer to increase dissolved oxygen and oxidation-reduction potential in a 1000-L treatment tank was tested at two water temperatures (~10°C and ~25°C) using both dechlorinated laboratory water and amended dechlorinated laboratory water. Water-only experiments were conducted and showed that dissolved oxygen increased with time in both dechlorinated laboratory water and amended dechlorinated laboratory water at both temperatures. Dissolved oxygen in ~10°C water had higher concentrations than in ~25°C water. ORP measurements conducted during testing with both water temperatures and types did not show a high degree of correlation in ORP values with increased treatment time. During treatment it was also determined that ozone and hydrogen peroxide were not produced as by-products. Biological effectiveness testing was completed with the algae, Selenastrum capricornutum and bacteria, Escherichia coli and Enterococcus faecium, in dechlorinated laboratory water. The system was found to be ineffective in the treatment of algae and bacteria when operated for approximately 30 minutes, and ineffective in treatment of algae when operated for 6 hours. However, when operated for 24 hours results suggest that the Kria Ionizer was effective against bacteria.LSRI-GWRC would like to thank EcoUSA, LLC (Denver, Colorado, USA) for their application to our laboratory-based testing program and for providing the Kria Ionizer Model DG-100 ballast water treatment (BWT) testing unit. Mike Mangham of EcoUSA, LLC provided installation and operational training and support prior to the start of and during testing. The results and conclusions contained in this report reflect the scientific data and opinions of the LSRI-GWRC research team. The technology developer did not review or provide comments on this report. This work was supported by the United States Maritime Administration (United States Department of Transportation, Washington, D. C.) and the United States Environmental Protection Agency’s Great Lakes Restoration Initiative

Tests of the Amglo Kemlite Bench-scale Ballast Water Management System

This technical report presents findings from bench-scale tests evaluating the performance of the Amglo Kemlite Bench-Scale Ballast Water Management System (BWMS), hereafter Amglo Kemlite BWMS, developed by Amglo Kemlite Laboratories Inc. of Bensonville, Illinois. The Amglo Kemlite BWMS applies a patented light separation technology to treat ballast water with pulsed poly-chromatic light. The light separation technology employs high-energy, pulsed xenon flash lamps. The Amglo Kemlite BWMS is a bench-scale prototype with a single production-sized lamp and power supply installed in an appropriately sized unit for laboratory evaluation

NBOT 2.5 HP Report

This technical report presents the bench-scale evaluation of the Nano Bubble Ozone Technology 2.5-horsepower unit (NBOT 2.5-HP) developed by NanoClear Group Inc. of Rockville, Maryland. This evaluation was the first to assess NBOT 2.5-HP as a potential in-tank, recirculating ballast water treatment method for the Laurentian Great Lakes. The evaluation began in September 2019 and ended in March 2020. All analyses occurred at the Lake Superior Research Institute (LSRI) at the University of Wisconsin-Superior (UWS) in Superior, Wisconsin, USA. The NBOT 2.5-HP uses cavitation to create ultrafine microbubbles (nanobubbles) containing ozone (O3) generated by the system. According to the developer, the resulting ozone and hydroxyl radical biproducts destroy all chemicals containing activated functional groups (aldehydes, ketones, amines, nitrates, etc.), RNA, DNA, peptides, steroids, as well as activated organic compounds (herbicides and pesticides), and microbial toxins. The ability of NBOT 2.5-HP to increase dissolved ozone in a 1,000-L treatment tank was tested at two water temperatures (~15°C and ~25°C) using both dechlorinated laboratory water (LW) and the more challenging amended dechlorinated laboratory water (LW-TMH). In LW, NBOT 2.5-HP increased ozone (<15 minutes) upon treatment and reached equilibrium after approximately 2 hours of treatment under both temperature conditions. In LW-TMH, no increase in ozone was observed initially upon treatment. Instead, ozone increased after approximately 2 hours and reached equilibrium after 5 to 7 hours under both temperature conditions. Degradation rates of dissolved ozone in LW and LW-TMH were examined at two water temperatures (~15°C and ~25°C). In general, ozone degradation rates were lower at 15°C than at 25°C while degradation occurred more rapidly in LW-TMH than in LW. Biological effectiveness tests examined the ability of NBOT 2.5-HP to induce mortality in biological organisms over time in both LW and LW-TMH. Three classes of organisms were tested: bacteria (Escherichia coli and Enterococcus faecium), green algae (Selenastrum capricornutum), and zooplankton (D. magna neonates, D. magna ephippia, and Eucyclops spp.). In LW, the algae and bacteria experienced 100% mortality, or no live organisms (a count of <1 MPN/100 mL) after 30 minutes of treatment. In LW, D. magna neonates and Eucyclops spp. experienced 100% mortality after 30 – 60 minutes of treatment. In LW-TMH, the algae and E. coli experienced 100% mortality, or no live organisms (a count of <1 MPN/100 mL) after 240 minutes of treatment. In LW-TMH, only one sample replicate had an E. faecium count of 3 MPN/100 mL at 240 minutes and no live organisms were detected after 390 minutes of treatment. In LW-TMH, D. magna neonates and Eucyclops spp. experienced 100% mortality after 240 – 390 minutes of treatment. In both water types, the D. magna ephippia had a hatch rate of 22.5 - 36% following treatment. These results demonstrate that NBOT 2.5-HP is effective at inducing mortality in a wide range of organisms within size classes regulated in ballast water discharge in two different water qualities. Chronic Residual Toxicity (CRT) testing examined the potential for water treated with NBOT 2.5-HP to cause toxicity to organisms in receiving water upon discharge. This testing was conducted using LW treated with the NBOT 2.5-HP system. Three classes of organisms were tested: green algae (Selenastrum capricornutum), zooplankton (Ceriodaphnia dubia), and vertebrate (Pimephales promelas). No statistically significant effects on growth, survival or reproduction were seen.LSRI-GWRC would like to thank the American Marine University Research Institute Inc. for their application to our laboratory-based testing program and for providing the Nano Bubble Ozone Technology 2.5 horsepower (NBOT 2.5-HP) ballast water treatment (BWT) testing unit. Brian Domrese of NanoPure Tech provided installation and operational training and technical support prior to the start of and during testing. Dr. Peter Moeller of the National Oceanic and Atmospheric Administration’s (NOAA) National Ocean Service (NOS) in Charleston, South Carolina provided technical advice and assistance during implementation of bench scale testing. This work was supported by the United States Maritime Administration (United States Department of Transportation; Washington, D. C.) and the United States Environmental Protection Agency’s Great Lakes Restoration Initiative

NBOT 3 HP Report

This technical report presents the bench-scale evaluation of the Nano Bubble Ozone Technology 3-horsepower unit (NBOT) developed by NABAS Group Inc. of Rockville, Maryland. This evaluation was the first to assess NBOT as a potential, in-tank, recirculating ballast water treatment method for the Laurentian Great Lakes. The evaluation began in March 2019 and ended in June 2019. All analyses occurred at the Lake Superior Research Institute (LSRI) at the University of Wisconsin-Superior (UWS) in Superior, Wisconsin, USA. The treatment technology uses cavitation to create ultrafine microbubbles (nanobubbles) containing ozone (O3) generated by the system. According to the developer, the resulting ozone and hydroxyl radical biproducts destroy all chemicals containing activated functional groups (aldehydes, ketones, amines, nitrates, etc.), RNA, DNA, peptides, steroids, as well as activated organic compounds (herbicides and pesticides), and microbial toxins. The ability of NBOT to increase dissolved ozone and oxidation-reduction potential in a 1000-L treatment tank was tested at two water temperatures (~10°C and ~25°C) using both dechlorinated laboratory water and amended dechlorinated laboratory water. Ozone levels observed to be generated by NBOT were lower than anticipated based on observations by Dr. Peter Moeller of the National Oceanic and Atmospheric Administration’s (NOAA) National Ocean Service (NOS) who was utilizing a newer model of NBOT. Biological dose effectiveness testing was not completed, per the developer’s request, due to below expected levels of ozone.Great Lakes Restoration Initiative; Maritime Administratio

B-QUA

This technical report presents findings from bench-scale verification tests evaluating the performance of the B-QUA Quick Ballast Water Monitoring Kit, hereafter B-QUA, in freshwater. B-QUA was developed by LuminUltra Technologies Ltd. of New Brunswick, Canada. Researchers began conducting the bench-scale evaluation in October 2019 and ending in February 2020 at the Lake Superior Research Institute (LSRI) of the University of Wisconsin-Superior (UWS) in Superior, Wisconsin, USA. The monitoring kit utilizes adenosine triphosphate (ATP) and size fractionation to quantify living organisms in marine, brackish, and freshwater. The measurement of ATP is one of the indicative analyses to test for gross compliance with the D-2 ballast water management standard under the International Maritime Organization’s (IMO) Ballast Water Management (BWM) Convention, which applies to countries outside of the United States, including Canada (IMO, 2015). Two phases of testing were done. Phase I testing was completed in two water types using cultured organisms in the three regulated size classes, utilizing the pathogen indicator organisms Escherichia coli and Enterococcus faecium, the algae Haematococcus pluvialis and the zooplankton Ceriodaphnia dubia and Daphnia magna. Phase II testing was completed using naturally occurring Great Lakes organisms in the Duluth-Superior Harbor of Lake Superior in two of the regulated size classes. Phase I testing showed high correlation of B-QUA’s luminometer output (i.e., cATP values) with microscopic counts for the algae Haematococcus pluvialis (≥10 μm to 50 μm size class) in both water types. However, the B-QUA system was unable to detect E. coli or E. faecium (<10 μm size class) at levels above the D2 regulatory value in either water type in Phase I. Phase II correlation between B-QUA cATP values and microscopic counts was good for natural assemblages of phytoplankton and zooplankton in Duluth-Superior harbor water.This work was supported by the United States Maritime Administration (United States Department of Transportation; Washington, D.C) and the United States Environmental Protection Agency’s Great Lakes Restoration Initiative