Water Research Foundation October 20, 2015- Plantsville, CT CYANOBACTERIA AND CYANOTOXINS : RESEARC HUPDATE Djanette Khiari, Ph.D., Water Research Foundation CWWA/CT SECTION AWWA,2015 FALL CONFERENCE October 20, 2015- Plantsville, CT

Foundation Background Research Cooperative Sponsor research Communicate knowledge Volunteers and utility involvement are key Primarily supported by utilities Some federal and partnership money

Western Lake Erie Algal Bloom (August 2014, NASA) Toledo, Ohio (population ~50,000) 1 µ/L microcystin in finished water Do Not Drink/Do Not Boil Advisory

Agenda Understand the problem Occurrence Toxicology Regulatory status/EFPA activities Available tools to manage cyanotoxin Event WRF projects highlights

Tool Box Analytical Methods Watershed Management Source Water Protection Source Water Monitoring/ Treatment In-Plant Treatment Options Public Outreach and Risk Communication

Select WRF Reports Release of Intracellular Metabolites from Cyanobacteria During Oxidation Processes ( Project 4406) Evaluation of Integrated Membranes for Taste-and-Odor and Toxin Control (Project 4016) International Guidance Manual for the management of Toxic Cyanobacteria (Project 3148) Reservoir Management Strategies fro Control and Degradation of Algal Toxins (Project 3123) Treating Algal Toxins Using Oxidation, Adsorption, and Membrane Treatment (Project 2839)

Recent WRF Projects Optimizing Conventional Treatment fro Removal of Cyanobacteria and Toxins (Project 4315) - published Management of Treatment Sludge Impacted by Cyanobacteria (Project 4523) – on going Treatment of Algal Toxins in Rivers and River-Influenced Groundwater (project 4526)-– on going Cyanotoxin Guide for Utility Managers (Project 4548)- on going

Background

Cyanobacteria Taste and odor problems Toxin production Photosynthetic prokaryotic organisms Contain blue pigment in addition to chlorophyll a Occur singly or in colonies in diverse habitats Very common around the world Can form surface blooms that adversely affect drinking treatment and supply due Filter clogging Operational costs Taste and odor problems Toxin production Prokaryotes organisms that lack a membrane bound cell nucleus

Factors Affecting the Formation of Algal Blooms

Occurrence of Cyanobacteria in the US Graham, 2007

microcystin-LR saxitoxin cylindrospermopsin On the right we have geosmin and MIB. Geosmin and MIB are responsible for the musty earthy types of odor. Actually , geosmin takes its name from the greek “geo”+ osme (odor) earth They can be smelled at very low concentrations (5 to 9 ng/L) On the left some examples of algal toxins anatoxin-a cylindrospermopsin © 2011 Water Research Foundation. ALL RIGHTS RESERVED.

Cyanotoxins and Properties Toxin Group Primary target organ in mammals Cyanobacterial genera Microcystins Liver, possible carcinogen in this and other tissues Microcystis, Anabaena, Planktothrix (Oscillatoria), Nostoc, Hapalosiphon, Anabaenopsis, Aphanizomenon Cylindrospermopsins Liver and possibly kidney. Possible genotoxic and carcinogenic Cylindrospermopsis, Aphanizomenon, Umezakia, Raphidiopsis, Anabaena Anatoxin-a(S) Nerve synapse Anabaena Saxitoxins Nerve axons Anabaena, Aphanizomenon, Lyngbya, Cylindrospermopsis

Regulations

What is a EPA Health Advisory “HAs serve as informal technical guidance for unregulated drinking water contaminants to assist Federal, State and local officials and managers of public or community water systems in protecting public health as needed. They are not to be construed as legally enforceable Federal standards."

Is HA a Defacto Regulation? HA is treated like an MCL by many, but it really it is an MCLG, because EPA did not consider technology feasibility and cost, and it did not have proposal and comment period

What are some states doing? Some enacting EPA HA and guidance as is Some enacting HA but using the 10 day period for a notice Some evaluated and postponed action—partially in response to AWWA comments Some ignoring –don’t want any data collected

Analytical Methods

Selectivity and Sensitivity Relationships AWWA Webcast Program: 5/19/04 Selectivity and Sensitivity Relationships between Analytical Methods for Microcystins 100 NMR LC/MS 75 Biological and biochemical l Selectivity HPLC 50 Physico-chemical The various methods for analysis and detection have different selectivity levels and sensitivity levels. For example ELISA is very sensitive but is not selective for the individual microcystins, while NMR is very selective but requires larger quantities for analysis. TLC 25 ELISA Bioassay PPIA u g ng pg Sensitivity

Analyzing Cyanotoxins) Microcystins Cylindrospermopsin Anatoxin-a ELISA1  PPIA2 Mouse bioassay Receptor-binding assay HPLC -DAD3 GC/MS4 LC/MS5 LC/MS/MS6 1Enzyme-linked immunosorbent assay 4Gas chromatography/mass spectrometry 2Protein-phosphatase inhibition assay 4Liquid chromatography/mass spectrometry 3High Pressure Liq uid Chromatography-Diode Array Detector 5Liquid chromatography/tandem mass spectrometry

ELISA versus LC/MS/MS Characteristics Measure groups of variants Measure individual variants Quantitation Semi-quantitative Quantitative Sample volume <0.5 ml MRL 0.15 µg/L 0.1 µg/L Turn-around time Fast longer Instrumentation Inexpensive costly Level of expertise Easy High Other Kits available commercially EPA Methods 544 (microcystins) and 545 (Cylindrospermopsin) Abraxis ADDA-specific kit

Source Water Management

Early Warning Early warning systems must be functioning continuously to be effective Identify early indicators (e.g. pH, water temperature, Secchi disk depth, location/extent of thermocline) Define trigger levels for increased monitoring Identify action thresholds that tie source water monitoring to operational decisions WHO Cyanobacteria Cell Count Action Levels that Trigger Toxin Sampling for Drinking Water Species Action Level Microcystis spp. 2,000 cells/mL Combination of all potentially toxic cyanobacteria spp. Present 15,000 cells/mL

Source Water Monitoring Approaches Parameters/Variables Monitoring Type Parameters/Variables Demands on equipment Who Basic   Minimal Site inspection for indicators of toxic cyanobacteria in waterbody Transparency, discoloration, scum formation Secchi disc, regular site inspection by trained staff; Operators Surrogates Low to moderate Potential for cyanotoxin problems in waterbody Total phosphorus, nitrate and ammonia, thermal stratification, transparency, cholorphyll Photometer, boat, depth sampler, Secchi disc, submersible temperature/oxygen probe; basic skills but requires specific training and supervision Limnologist Cyanobacteria In waterbody and drinking water Dominant taxa (quantity): determination to genus level only is often sufficient; quantification Microscope, photometer is useful; specific training and supervision is required, but quite easily achieved Phycologist - trained by phycologist Cyanotoxins Moderate to High Microcystin, anatoxin-a, cylindrospermopin, ELISA kits (moderate); LC/PDA (moderately high); LC/MS (high) Chemist

Management of Cyanobacteria: Summary of Options Intervention Type Technique Physical Control Selective intake Mixing - destratification Dilution to decrease retention time Nutrient Control External Watershed management Internal Aeration & mixing Oxygenation (hypolimnetic) Sediment “capping” with P-binding agents e.g. Modified clays, Alum Chemical Control Non-Chemical Control Algaecides Novel Technology (e.g. ultrasound) Biomanipulation Viruses, bacteria, exotic algae Biological Control

In- Plant Treatment Options

Intracellular Cyanotoxins Removal (Intact Cells) General Rule: Avoid pre-oxidation that lyses cells Removing intact cells is: more cost effective than chemical inactivation/de removes a higher fraction of DBP precursors removes a higher fraction of intracellular T&O compounds it is easier to monitor removal. Coagulation/Sedimentation/Filtration Effective for the removal of intracellular/particulate toxins. Membranes Microfiltration and ultrafiltration are effective at removing intracellular/particulate toxins. Typically, pretreatment is used. Flotation Flotation processes, such as Dissolved Air Flotation (DAF), are effective for removal of intracellular cyanotoxins since many of the toxin-forming cyanobacteria are buoyant.

Extracellular Cyanotoxins Removal Potassium Permanganate Membranes Typically, NF has a molecular weight cut off of 200 to 2000 Daltons; individual membranes must be piloted to verify toxin removal. RO is effective. Potassium Permanganate Effective for oxidizing microcystins and anatoxins. Not effective for cylindrospermopsin and saxitoxins. Ozone Very effective for oxidizing extracellular microcystin, anatoxin-a and cylindrospermopsin. Chloramines Not effective. Chlorine dioxide Not effective with doses typically used for drinking water treatment. Chlorination Effective for oxidizing extracellular cyanotoxins as long as the pH is below 8, ineffective for anatoxin-a. UV Radiation Effective at degrading toxins but at impractically high doses. Activated Carbon PAC/GAC: Most types are generally effective for removal of microcystin, anatoxin-a, saxitoxins and cylindrospermopsin. Mesoporous carbon for microcystin and cylindrospermopsin.

Distribution Systems Integrated Treatment systems EC: extracellular © 2011 Water Research Foundation. ALL RIGHTS RESERVED.

Management Of Treatment Sludge Impacted By Cyanobacteria Optimizing Conventional Treatment for Removal of Cyanobacteria and their Metabolites (WRF Project 4315) Management Of Treatment Sludge Impacted By Cyanobacteria

Key Results Although removal of cyanobacteria through conventional coagulation can be very effective, 100% cell removal is unlikely in normal full scale operations. In the event of high cell numbers entering the plant monitor for cell carryover and accumulation in clarifiers, this can lead to serious water quality problems if not rectified

Cell number and turbidity variations with alum dose, A. circinalis in R. Murray water

Key Results Once captured in the sludge, cyanobacteria can remain viable and possibly multiply over a period of at least two – three weeks. Simultaneously, within one day some cells in the sludge will lyse and release NOM and metabolites. Toxin release from sludge can be up to 5 times the initial concentration. The risk associated with supernatant return may be higher than previously assumed 

* Dependent on the GAC remaining adsorption capacity Summary of risks associated with supernatant recycling based on different cyanobacteria and water treatment process efficiencies. VH-very high; H-high; M-medium; L-low cyanobacteria Pseudanabaena D. circinale M. species C. raciborskii   Metabolite MIB GEO STX MCs CYN WQ Risk from supernatant recycling VH M H Treatment barrier Residual risk associated with individual treatment barriers PAC L Coagulation Ozone M/L GAC (physical removal)* M/H GAC (biological removal)** H/M Chlorine CT>50 mg L-1 min-1 * Dependent on the GAC remaining adsorption capacity ** Dependent on the presence of degrading bacteria in the biofilm

Treatment of Algal Toxins in Rivers and River Influenced Groundwater WRF Project 4526

4526: Project Goals Establish practical guidelines for operators Investigate the effectiveness Chlorine Chlorine dioxide Potassium permanganate Powdered activated carbon adsorption Ozonation Oxidants in combination with chlorine Evaluate the potential to use geosmin and MIB removal as indicators of the removal of specific algal toxins

Effect of Ozone in High pH Water

Effect of PAC in High pH Water

Bench-Scale Summary Initial concentration does not effect removal Taste and Odor compounds do not effect removal MC-LR easier than T&O for ozone MC-LR about same as T&O for PAC PAC dose of 10-20 mg/L effectively reduces or eliminates MC-LR Cl2 dose of 4 mg/L can remove 40-80% of MC-LR KMnO4 dose of 1.5 mg/L removes ~80% of MC-LR Ozone dose of 2 mg/L highly effective for removal

Cyanotoxin Utility Action Guide and Research Needs WRF Project 4548

Research Partner: AWWA/WITAF Phase I: A Water Utility Manager’s Guide to Cyanotoxins: Published Phase II: Managing Cyanotoxins in Drinking Water: A Technical Guidance Manual for Drinking Water Practitioners

Quick Self-Assessment Step 1: How Vulnerable is Your Water Utility to a Cyanotoxin Problem?   High Medium Low None Source Water Monitoring Do you have a source water monitoring program in place? Don’t monitor our source water before treatment Test our water some (e.g., turbidity, total organic carbon) as it enters treatment plant Monitor source water monthly (e.g., chlorophyll a, algae counts) at different depths and locations Have a comprehensive source water monitoring program, sampling at least weekly at different depths, locations Does your source water quality monitoring program evaluate changes over the year? No Yes, tracks monthly water quality trends (e.g. to help us decide which source(s) to use) Yes, tracks trends at least weekly of all monitored parameters Do you track changes by comparing water quality data from year to year? Yes, seasonal or annual averages are tracked and compared Yes, charts are created with monthly data for at least the last five years

Quick Self-Assessment Step 1: How Vulnerable is Your Water Utility to a Cyanotoxin Problem?   High Medium Low None Source Water Quality and Aesthetics Do you have algae growth in your source water? Yes, we have blooms and add copper sulfate regularly Yes but we don’t have to adjust treatment in response We have minor algae growth but no visually obvious blooms Very minimal, if any, growth Does your source water stratify thermally in the summer? Yes, strong thermocline and turnover in late summer/fall with noticeable water quality changes Stratifies but no noticeable changes in water quality with turnover Stratifies some during the day but mixes at night No Is your surface water source affected by drought? Yes water level drops, water is warmer due to drought conditions Water level drops a small amount, no water temperature increases Does your water look green or blue-green? Yes, the color of our reservoir changes noticeably Slightly

Quick Self-Assessment Step 1: How Vulnerable is Your Water Utility to a Cyanotoxin Problem?   High Medium Low None Cyanobacteria in the Treatment Process Do you have treatment processes that are exposed to sunlight? Yes, most of our unit processes are outdoors and uncovered Yes at least one of our unit processes is exposed to sunlight No Is your filter backwash green? Yes, frequently Yes, periodically Do you have taste and odor problems? Yes we frequently get complaints during the summer Yes we periodically get complaints Once every few years Are your basins regularly cleaned? No, never Maybe once every few years At least once a year More than once a year

Quick Self-Assessment Step 2: How Many Tools Are In Your Toolbox?   Yes No If yes, has the measure been evaluated for addressing cyanotoxins? Water Quality Management/Treatment Algae reduction tools for source (e.g. raw) water supply, including Enhanced circulation/mixing Chemical addition (e.g. copper sulfate, chlorine) Ultrasound Other Ability to select from different intakes, both in terms of depths/locations and time (i.e. being able to switch intakes without delay or much effort) Intake Inline Chemical Addition Permanganate Chlorine Chlorine dioxide Conventional Treatment Membrane Filtration Oxidative Processes (being used for DBP precursor removal, taste and odor control, or other chemical contaminant removal) Ozone Peroxide Disinfection Processes Activated Carbon (powdered or granular) or other adsorptive media Biological Activity

Quick Self-Assessment Step 2: How Many Tools Are In Your Toolbox?   Yes No If yes, has the measure been evaluated for addressing cyanotoxins? Supplying Water Can provide enough treated water to all consumers with surface water supply out of service for a week. If yes, would not feel restricted by the cost of using or treating the alternative supply at least once a year. Communicating with the Public Well-organized, exercised public notification program Established communication network with the local public health and medical community

Public Outreach and Communication Before Event – Preparing for an Advisory Consider timing, audience(s), channels, messages, approval procedures Collaborate with Partners Develop Message Conduct Exercises   During Event – Issuing an Advisory Implement established procedures for Initiating an Advisory Distributing an Advisory Ending an Advisory After Event – Evaluating an Advisory  Reporting Requirements  Debrief Event  Conduct an Evaluation  Modify SOPs  Update Public Outreach Procedure

Take Home Messages Know your system Be technically prepared Early warning Know how to manage source and adjust treatment Be ready to communicate With the public With the press

Important Aspects of the HAHA How will utilities communicate a children hA vs adults? Recommendations document how to balance an ELISA result vs LC?MS?MS ? resampling allowed-how time this and actions-24 hr resample notify public within 24 hrs lift advisory after 3 consecutive < 0.3

www.asdwa.org/habs State Webpage Title Web Link California Blue-Green Algae (Cyanobacteria) Blooms http://www.cdph.ca.gov/healthinfo/environhealth/water/pages/bluegreenalgae.aspx Illinois Harmful Algal Blooms (HABs) and Algal Toxins http://www.epa.state.il.us/water/algal-bloom/index.html Indiana Addressing Concerns About Blue-Green Algae http://www.in.gov/idem/algae/ Maryland Harmful Algae Blooms in Maryland http://www.dnr.state.md.us/bay/hab/index.html Massachusetts Algae Information http://www.mass.gov/eohhs/gov/departments/dph/programs/environmental-health/exposure-topics/beaches-algae/algae-information.html Nebraska 2013 Toxic Blue-green Algae and Bacteria Sampling Results http://www.deq.state.ne.us/Beaches.nsf/LakeSampling13 New Hampshire Recreational Exposure to Cyanobacteria (Blue-Green Algae) http://des.nh.gov/organization/divisions/water/wmb/beaches/cyano_bacteria.htm Ohio Harmful Algal Blooms: Information for Public Water Systems http://epa.ohio.gov/ddagw/HAB.aspx Oregon Algae Resources for Drinking Water http://public.health.oregon.gov/HealthyEnvironments/DrinkingWater/Operations/Treatment/Pages/algae.aspx Vermont Cyanobacteria: Blue Green Algae http://healthvermont.gov/enviro/bg_algae/bgalgae.aspx Washington Blue-Green Algae http://www.doh.wa.gov/CommunityandEnvironment/Contaminants/BlueGreenAlgae.aspx www.asdwa.org/habs

Cyanotoxin Drinking Water Advisory Thresholds State Microcystin Anatoxin A Cylindrospermopsin Saxitoxin Florida 10 µg/L, based on Microcystin-LR only None Ohio 1 µg/L, based on Microcystin-LR but meant to include all microcystin congeners 20 µg/L 1 µg/L 0.2 µg/L Oklahoma 1 µg/L with a goal of non-detect, based on all microcystin congeners Oregon Microcystin should be below 1 µg/L in finished water, otherwise a DO NOT DRINK public notice will be posted. Based on all microcystin congeners 3 µg/L Minnesota 0.04 µg/L, based on Microcystin-LR but meant to include all microcystin congeners Quebec 1.5 µg/L expressed as Microcystin-LR toxic equivalents including congeners LA, RR, YR, and YM 3.7 µg/L1 1 Canadian drinking water advisory threshold Source: Survey of state drinking water administrators conducted by the Association of State Drinking Water Administrators (ASDWA), D. Mason correspondence

EPA RECOMMENDS STRATEGIC PLAN

Thank You Comments or questions, please contact: Djanette Khiari dkhiari@waterrf.org For more information visit: www.waterrf.org