1. 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, Plantsville, CT

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

3. 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

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

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

6. 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)

7. 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

8. Background

9. 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

10. Factors Affecting the Formation of Algal Blooms

11. Occurrence of Cyanobacteria in the US
Graham, 2007

12. 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.

13. 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

14. Regulations

15. 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."

16. 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

18. 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

19. Analytical Methods

20. 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

21. 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

22. 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

23. Source Water Management

24. 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

25. 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

26. 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

27. In- Plant Treatment Options

28. 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.

29. 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.

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

31. 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

32. 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

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

34. 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 

35. * 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

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

37. 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

38. Effect of Ozone in High pH Water

39. Effect of PAC in High pH Water

40. 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 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

41. Cyanotoxin Utility Action Guide and Research Needs
WRF Project 4548

42. 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

45. 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

46. 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

47. 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

48. 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

49. 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

50. 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

51. 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

52. 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

53. www.asdwa.org/habs State Webpage Title Web Link California
Blue-Green Algae (Cyanobacteria) Blooms
Illinois
Harmful Algal Blooms (HABs) and Algal Toxins
Indiana
Addressing Concerns About Blue-Green Algae
Maryland
Harmful Algae Blooms in Maryland
Massachusetts
Algae Information
Nebraska
2013 Toxic Blue-green Algae and Bacteria Sampling Results
New Hampshire
Recreational Exposure to Cyanobacteria (Blue-Green Algae)
Ohio
Harmful Algal Blooms: Information for Public Water Systems
Oregon
Algae Resources for Drinking Water
Vermont
Cyanobacteria: Blue Green Algae
Washington
Blue-Green Algae

54. 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

55. EPA RECOMMENDS STRATEGIC PLAN

56. Thank You Comments or questions, please contact: Djanette Khiari
For more information visit: