Funded Projects

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The Use of Cultivatable Seaweeds to Mitigate HABs Caused by Alexandrium and Pseudo-nitzschia and Minimize the Accumulation of Toxins in Bivalves

Institution: Stony Brook University
Location: New York
Total Funding: $200,000
Project Period: September 2024-August 2025
Project Summary: This project investigates how native US seaweeds—Saccharina Latissima, Ulva spp., and Gracilaria tikvahiae— can be used to restrict two common marine harmful algal blooms (HABs), Alexandrium and Pseudo-nitzschia, and reduce toxin levels in bivalves. Experiments will be conducted in 300 L mesocosms and1,500 L flow-through raceways, to evaluate how seaweed concentrations and water exchange rates impact HAB proliferation and toxin accumulation in blue mussels. Researchers hope these large-scale experiments will provide a better understanding of how this approach would work in a marine ecosystem. Given that seaweeds are a bourgeoning aquaculture crop in the US, this HAB control technology has the potential to be revenue-generating while concurrently protecting aquacultured shellfish against HABs and their toxins.

Turmeric Triumph: Unveiling Curcumin’s Power in Controlling Alexandrium and Pseudo-nitzschia and Their Toxins

Project Principal Investigators: Cynthia Heil, Ph.D., Emily Hall, Ph.D.
Institution: Mote Marine Laboratory
Location: Sarasota, FL
Total Funding: $157,616
Project Period: September 2024-August 2025
Project Summary: Mote Marine Laboratory aims to investigate the phenolic pigment Curcumin, derived from turmeric, as a potential tool for removal of the dinoflagellate Alexandrium spp. and the diatom Pseudo-nitzschia spp. and their associated toxins. Curcumin has been successfully shown to mitigate both cells and toxins of the Florida red tide dinoflagellate Karenia brevis in previous studies conducted at Mote Marine Laboratory. The goals of these bench-top scale experiments are 1. to assess Curcumin’s ability to minimize these species cell concentrations along with their associated toxins, 2. determine the optimal dosing of Curcumin in algal-affected water for maximum cell and toxin reduction, and 3. to determine the impact of Curcumin on water quality using standardized methods to measure pH, oxygen concentration, and nutrient concentrations.

High-throughput Screening to Identify Algicidal Bacteria and Compounds Active Against Alexandrium and Pseudo-nitzschia

Project Principal Investigator: Kirstie Francis, Ph.D.
Institution: Mote Marine Laboratory
Location: Sarasota, FL
Total Funding: $111,611
Project Period: September 2024-August 2025
Project Summary: This project aims to identify Marine Natural Products (MNPs) capable of algicidal activity using the Mote Microbial Library. High-throughput screening techniques will be used to identify algicidal agents capable of reducing harmful algal blooms (HABs). The project specifically targets the dinoflagellate Alexandrium spp. and the diatom Pseudo-nitzschia spp. as they pose significant threats to marine life along the East and West coasts of North America. . The goals of this project are to conduct a high-throughput screening of 1) the Mote Microbial Library to identify bacteria capable of direct algicidal activity and 2) the pre-fractionated extract library to identify MNPs from bacteria with algicidal activity against harmful algae species Alexandrium and Pseudo-nitzschia. Bacterial strains and pre-fractionated extracts which reduce algal cell counts by ≥ 50% will be further investigated to identify the strain using 16S sequencing or determine the compound responsible for activity via bioassay guided fractionation. Researchers hope this will provide more insight into potential biological control mechanisms for these HABs, paving the way for further research

Evaluation of TAML®/Hydrogen Peroxide Catalysis for Microcystis aeruginosa Control

Project Principal Investigator: Terrence Collins, Ph.D.
Institution: Carnegie Mellon University
Location: Pittsburg, PA
Total Funding: $197,648
Project Period: September 2024-August 2025
Project Summary: Harmful algal blooms (HABs) caused by Microcystis spp., express potent toxins that present significant threats to aquatic life and ecological health overall. In search of an economical and environmentally sustainable solution, this study aims to assess the efficacy of TAML® activators combined with hydrogen peroxide (H₂O₂) as an algicidal compound. TAML activators are bioinspired, small-molecule peroxidase mimics that activate hydrogen peroxide (H2O2), down to single digit ppm concentrations, to oxidatively degrade organics in water. This study has three primary goals utilizing tiered research approaches: 1) Tier 1 will identify the optimal dosage of TAML®/H₂O₂ to lower M. aeruginosa cell count/toxin levels at pH values conducive to algal blooms,, 2) Tier 2 will assess the effectiveness of TAML®/H₂O₂ catalysis to reduce naturally sourced blooms in mesocosm-scale assays to better understand the potential effects on an environmental scale, and 3) will evaluate the effect of TAML®/H₂O₂ on multicellular freshwater species Ceriodaphnia dubia (water flea) and Pimephales promelas (fathead minnow) to observe non-target effects.

Remediation of Alexandrium, Pyrodinium, and Karenia along with their Toxins by PAC-Modified Biochar

Project Principal Investigator: Toufiq Reza, Ph.D.
Institution: Florida Institute of Technology
Location: Melbourne, FL
Total Funding: $200,000
Project Period: September 2024-August 2025
Project Summary: This study aims to use biochar modified with poly-aluminum chloride (PAC) to flocculate HAB cells, Alexandrium, Pyrodinium, and Karenia, and absorb their associated toxins. Previous studies have shown PAC-modified clay to be effective in flocculating HAB cells, and biochar to adsorb HAB toxins. This research will synthesize modified biochars with PAC in various methods and will test their efficacies for simultaneous flocculation of various HAB cells and associated toxins. The objectives of this study are: 1) identify the best way to combine these two products so that both HABs and their toxins can be removed from water systems, 2) Determine the optimal concentrations for removal of HAB species Alexandrium, Pyrodinium, and Karenia using benchtop experiments, 3) conduct 80L vertical tank assays to better assess HAB cell abundance, toxin concentration, and photo-physiological response on a large scale. If successful, this research study could revolutionize HAB mitigation techniques and benefit marine ecosystems on a large scale.

Optimizing Clay Formulations for HAB Treatment Efficacy and Regulatory Approval

Project Principal Investigator: Donald M. Anderson, Ph.D.; Keith Ervin; Cynthia Ann Smith
Institution: Woods Hole Oceanographic Institution, K E Technologies LLC, Conn & Smith, Inc.
Location: Woods Hole, MA
Total Funding: $126,125
Project Period: September 2024-August 2025
Project Summary: Clay flocculation is a widely studied technique used to combat HABs. China and Korea have successfully implemented clay flocculation on large scales. This project aims to analyze two clay types, ingredient combinations, and costs to develop the most effective clay formulations that will be compliant with United States Federal regulations. A matrix of clay formulations will be developed to test effectiveness against Karenia brevis cells and associated toxins after active ingredients have undergone regulatory evaluations and efficacy assessments. After the clays and ingredients have been narrowed down to those with the simplest and most promising regulatory pathways, that subset of clays and ingredients will be combined and tested against Karenia brevis cells and associated toxins . Based on lab results and regulatory limitations, researchers will then draft pre-submission consultation request documents to the EPA for the optimal formulation and ingredients.

Nanobubbles-enabled foam fractionation for efficient removal of algae and algogenic pollutants

Project Principal Investigator: Wen Zhang, Ph.D., P.E., BCEE
Institution: New Jersey Institute of Technology
Location: Newark, NJ
Total Funding: $200,000
Project Period: November 2023 - October 2024
Project Summary: This project will evaluate and validate the technical and economic viability of foam fractionation for removing freshwater algae using air nanobubbles to produce bubble foam (a foamy structure of tiny bubbles usually stabilized by surfactants or surface-active agents and thus exhibits high surface areas and stability in water). The produced foam will adsorb water pollutants including harmful algal cells and their toxic metabolites via a foam fractionation separation process. There are only a few limited studies that report the use of nanobubbles in foam fractionation for algal separation and algogenic pollutant removal, despite the studies for other pollution remediation (e.g., PFAS) and biomolecular separation. The goals of this project are 1. To investigate different variables including surfactant types, on the effectiveness of algal separation, 2. Compare the algal separation efficiencies of nano-bubbles with and without surfactants on algogenic pollutants and 3. Investigate the separation mechanisms and optimization of removal kinetics. This project will support Ph.D. student, Yihan Zhang, and a postdoc researcher to work with the PI, Dr. Wen Zhang.

Developing a highly selective molecular tool for Microcystis aeruginosa to inhibit the production of microcystin from harmful algal blooms

Project Principal Investigator: Scott Hamilton-Brehm, Ph.D.
Institution: Southern Illinois University
Location: Carbondale, IL
Total Funding: $194,083
Project Period: November 2023 - October 2024
Project Summary: Antisense oligonucleotide (ASO) technology can be leveraged for targeted knockdown of undesirable mRNA gene products, with limited environmental disturbance. The use of exogenous ASOs has been well described in model prokaryotic organisms such as E. coli, S. aureus, and Pseudomonas sp., but ASO effectiveness on cyanobacteria is unknown. This project will study the use of ASOs to target and stop the production of the toxin microcystin by cyanobacteria at the molecular level. The objectives of this study are: 1. Develop an enhanced molecular delivery system for ASO’s that selectively targets Microcystis aeruginosa, 2. Evaluate the effectiveness of the molecular delivery system into M. aeruginosa cells in scaling culture volumes, 3. Evaluate environmental impact of the molecular delivery system on microbial, phytoplankton, and zooplankton communities.

Effective treatment concentrations of TAML®/H2O2 on Karenia brevis and brevetoxin removal

Project Principal Investigator: Terrence Collins, Ph.D.
Institution: Carnegie Mellon University
Location: Pittsburgh, PA
Total Funding: $178,981
Project Period: November 2023 - October 2024
Project Summary: TAML®/H2O2 has already been shown to degrade a wide variety of persistent water contaminants, including cyanotoxins and other micropollutants (MPs). Additionally, TAML®/H2O2 decays in use and does not induce adverse effects on zebrafish and mice. It is hypothesized that TAML®/H2O2 will inactivate Karenia brevis cells as well as destroy waterborne brevetoxins as both are vulnerable to oxidation. This project aims to perform Tier 1 and Tier 2 testing to determine if TAML®/H2O2 is effective for controlling K. brevis cells and associated brevetoxins and if treatment is ecologically safe for marine and estuarine species. The objectives are: 1. Determine the effective treatment concentrations and conditions using 2.8 L Fernbach flasks, 2. Find the relationship between treatment concentration and duration to cell number and toxin concentration, 3. Test the chosen treatment condition at a larger scale, using 2,000 L mesocosms, 4. Run EPA-certified acute toxicity tests using chosen treatment conditions at Marinco Bioassay Laboratory.

Targeted Destruction of Harmful Algal Blooms using Hybrid Materials for Flocculation, Sinking and Toxin Mitigation

Project Principal Investigator: Vijay John, Ph.D.
Institution: Tulane University
Location: New Orleans, LA
Total Funding: $150,000
Project Period: November 2023 - October 2024
Project Summary: Modified clay is a well-established technology of flocculation and sinking of HABs and activated carbon is known to remove toxins from the water column. This project proposes to encapsulate algaecide into widely available natural clay nanotubes known as halloysites and coat the halloysites in carbon. This targeted destruction with flocculate and kill HABs, while also reducing toxin exposure to off target organisms. The halloysites have been shown to flocculate and sink Karenia brevis from the water column through previously funded research. This work will expand to add the carbon and algaecide to the technology and test this technology on the freshwater HAB species, Microcystis aeruginosa. The objectives of this study are: 1. Develop the hybrid system of HNT nanotubes with attached algaecide and a coating of carbon, 2. Identify the effective dose of the algaecide filled nanotubes on M. aeruginosa and K. brevis by conducting flocculation and settling studies, scaling up from 20 ml, 1-L, to 80-L systems, 3. Assess the efficacy of the material for removal and degradation of the algae and toxins by measuring toxins (ELISA), measuring photosynthetic efficiency (PAM fluorometry), cell abundance/morphology (microscope/FlowCam), and cell viability (Sytox/Neutral red assay), and 4. Test the technology on field samples of a Microcystis bloom. This system will involve flocculation, sinking and targeted destruction of harmful algae with minimal off-target impact potentially generated by both the algaecide and the generated toxin.

Mitigation of Harmful Algal Blooms and Breakdown of Microcystin Using Electric Fields: Mesoscale Validation and Demonstration

Project Principal Investigator: Sudhir Sastry, Ph.D.
Institution: The Ohio State University
Location: Columbus, OH
Total Funding: $140,000
Project Period: November 2023 - October 2024
Project Summary: Moderate Electric Field (MEF) technology has a history in food processing as it uses less than 1,000 volts per centimeter to inactivate bacteria by causing pores in their cell membranes. This project will test the efficacy and scalability of using MEF combined with microcystin-degrading bacteria, Sphingomonas sp. to a) inactivate cyanobacteria; and b) to degrade microcystin. The combined technology will be tested using mesocosms of bloom-affected lake water. Additionally, the microbial community will be analyzed pre and post-treatment to assess how the technology affects non-targeted species.

NeroPure BNE against Microcystis aeruginosa

Industry: NeroPure LLC
Location: Redkey, IN
Total Funding: $56,000
Project Period: November 2023 - October 2024
Project Summary: NeroPure LLC. will be conducting mesocosm trials with Mote Marine Laboratory to study the efficacy of their proprietary liquid all-natural water restoration solution, NeroPure BNE. The study will consist of product application to sourced water with high cyanobacteria populations and toxins present to measure the product's impact in improving the overall ecology of the water system as well as the product's ability to manage high nutrient loads. The mesocosm trials will measure the cyanobacteria concentrations, toxins, and nutrients after NeroPure BNE application. The mode of action of NeroPure BNE on the microbial community composition will also be studied. The replicate trial experiment will help validate NeroPure BNE as an all-natural alternative solution to manage adverse impacts of eutrophication and the detrimental effects of high concentration cyanobacteria blooms.