Zequanox provides a non-chemical alternative to invasive mussel control

1. Managing Mussel Shell Debris with a New
Chemical Alternative
Sarahann M. Rackl, Ph.D., P.E.
Director of Water Technologies
Marrone Bio Innovations

2. Mussel Control Challenges Exacerbated by
Unrelenting Population Expansion
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3. Invasive Mussel Control Methods Must Consider
Shell Debris Management
• Shells from dead adult mussels
can clog screens or small
tubing causing similar impacts
as live mussel colonization
• Chemical solutions kill all the
mussels at the same time,
releasing masses of debris into
the treated system
• Regular treatments can help
mitigate impact of shell debris
• Settlement prevention
requires almost 24/7
continuous application
• More sustainable alternative
needed with flexibility to
improve shell debris
management
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4. Natural Product Chemistry
• Safer alternatives to standard
chemicals to protect public health
and the environment
– Products are more target specific
– Decreased potential for population resistance
• Typically not a toxic/contact mode of action
providing a more manageable control of pests
• Well established technology in
the pharmaceutical and agricultural industries
– Bacillus thuringiensis (israelensis) used to control black fly larva
in open waters - globally
– Production of antibiotics and antifungal agents effective against
diseases in crops
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5. Product Overview
• Derived from soil microbe
(Pseudomonas fluorescens CL 145A)
– Discovered by NYSM
– Composed of 100% dead cells
• Controls mussels in all life stages
• Perceived as food source—destroys the
mussel’s digestive system
– Causing slower mortality over time, not
all at once
• Highly selective toward zebra and
quagga mussels
• Effective in a broad range of water
conditions and temperatures
• Noncorrosive and nonvolatile
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6. Extensive Ecotox Studies Show No Impact to
Other Aquatic Species
MOLLUSCS
FISH Blue Mussel (Mytilus edulis) *
Bluegill sunfish (Lepomis macrochirus) Freshwater Mussel - Duck Mussel (Anadonta)
Channel catfish (Ictalurus punctatus) Freshwater Mussel - Black Sandshell (Ligumia recta)
Chinook Salmon (Oncorhynchus tshawytscha) Freshwater Mussel - Fatmucket (Lampsilis siliquoidea)
Coaster brook trout (Salvelinus fontinalis) Freshwater Mussel - Pink mucket (Lampsilis abrupta)
Common Carp (Cyprinus carpio) Freshwater Mussel - Hickorynut (Obovaria olivaria)
Fathead Minnow (Pimephales promelas) * Freshwater Mussel - Higgins Eye (Lampsilis higginsii)
Klamath Suckers (Catostomus sucker spp) Freshwater Mussel - Mucket (Actinonaias ligamentina)
Lake sturgeon (Acipenser fulvescens) Freshwater Mussel - Paper Pond Shell (Utterbackia imbecillis)
Largemouth bass (Micropterus salmoides) Freshwater Mussel - Plain Pocketbook (Lampsilis cardium)
Rainbow Trout (Oncorhynchus mykiss) * Freshwater Mussel - Washboard (Megalonaias nervosa)
Sacramento Splittail (Pogonichthys macrolepidotus) Freshwater Snail (Lymnaea peregra)
Smallmouth bass (Micropterus dolomieu)
Striped Bass (Morone saxatilis)
Walleye (Sander vitreus)
Yellow perch (Perca flavescens)
PLANTS AND ALGAE
Algae *
OTHERS Bindweed (Convolvulaceae)
Mallard Duck * Common Water Plantain (Alisma subcordatum)
Midge (Chironomidae) Curly Dock (Rumex crispus)
Mayfly (Baetis) Mallow (Malvaceae)
Amphipod (Hyalella azteca) * Nightshade (Solanaceae)
European Freshwater Crayfish (Austropotatamobius pallipes) Smallflower Umbrella Sedge (Cyperus difformis)
Freshwater Crustacean (Asellus aquaticus)
Freshwater Water Flea (Daphnia magna) *
* EPA required
Studies completed or will receive final report in 2012. Conducted by Institute of Technology, Sligo, Ireland;
6 New York State Museum and USGS; U.S. Bureau of Reclamation; Certified Good Laboratory Practices (GLP) Lab; and MBI lab

7. Treatment Programs Designed to Mitigate Shell
Debris Impacts
Periodic Preventative
Designed for facilities with tolerance Goal is to limit the number of mussels
for moderate to large shell sizes (larger that exceed 4mm in size
than 4 mm in size) Ideal for sensitive systems and
Treatments occur 1 -3 times annually equipment
Modification - slower/reduced kill for Treatments are performed on a
large populations can provide regular basis (approx. every 4-6 wks)
additional mitigation Modification – More frequent
Monitoring Level of Control – treatments to further reduce shell size
Quantified by percent of mussel s that Monitoring Level of Control–
die over time Evaluation of total biomass on
settlement plates and confirmation of
no mussels > 4 mm
• Product applied with standard equipment
• Treatments can be completed within hours (~6 hrs)
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8. Methods to Assess Treatment
• Bioboxes connected to facility’s
service water system
– Used for pilot demonstration (direct
application) and facility treatments
• Mussels collected locally for assessment
of periodic treatments or settlement
plates are inserted for assessing
preventative treatments
• Configured for comparison
between treated and control
• Product measured as turbidity
(NTU) during treatments
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9. Equivalent Level of Efficacy Observed Throughout
North America
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10. Case Study: Bureau of Reclamation’s Davis Dam
• Tested both Periodic and Preventative
treatment programs in 2011
• Bullhead City, AZ; Colorado River
• Five turbine, 240 MW hydropower
facility
• Each turbine has a separate cooling
water system
• 5000 gpm per cooling water subsystem
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11. Davis Dam Biobox Monitoring Schematic
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12. Low Dose Treatment Programs Provide Additional
Debris Management Over Time
90.0
80.0
70.0
60.0
Mortality (%)
50.0
Treated mean mortality
40.0 Control mean mortality
30.0
20.0
10.0
0.0
0 10 20 30 40 50 60 70 80 90 100 110 120
Time (days)
Initial treatment occurred at 0 days; blue arrows indicate timing
of additional lower concentration treatments which had a
12 cumulative effect on mussel mortality over time

13. Biomass Control at Mid-System Locations
0.30
Settlement Biomass (dry g/plate)
0.25
0.20
Treated Control
Settlement in Treated System at
0.15
Mid-System Location
0.10
0.05
0.00
Midpoint
• Demonstration of the preventative
treatment program
• 91.2% less biomass based on total plate
Settlement in Control System at biomass
Mid-System Location

14. Biomass Control at Outlet Locations
0.30
Settlement Biomass (dry g/plate)
0.25
Treated Control
0.20
0.15
0.10
Settlement in Treated System at Outlet
Location
0.05
0.00
Outlet
• 79.7% less biomass in the treated system
• Settlement analyzed three weeks after final
Zequanox treatment; new growth not excluded
during three-week period
Settlement in Control System at
Outlet Location

15. Summary of Davis Dam 2011 Case Studies
• Low concentration repeated treatments on adult mussels can
achieve accumulated mussel mortality over time yet reduce
influx of shell debris
• Treatments can achieve equivalent mussel control to chemical
treatments without causing shell debris to enter the system
all at once
• Successive low concentration treatments on recently settled
mussels can achieve a high level of control resulting in a
reduction of mussel biomass growth (Preventative Treatment
Program)
• Significant level of mussel control during extreme algae bloom
events
• Treatments caused no impact on Colorado River water quality
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16. Other Benefits of Zequanox Treatments
• Minimal use restrictions
• Elimination of quenching/detoxification before discharge
• No risk of release of toxic, hazardous chemicals into the
environment
• Minimal to no discharge monitoring requirements
• Low risk to employees
– Only minimal PPE needed
• No risk of corrosion or equipment damage
• Short treatment times
• Minimal impact on process and operations
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17. Marrone Bio Innovations
MBI discovers, develops, and markets
effective and environmentally responsible
natural products (biopesticides) that fill
unmet needs for weed, pest and plant
disease management.
We have our own microbial natural product
discovery screening
We in-license proprietary technology and
products (plant extracts and microbes), such as
Pf CL145A from NYSM (Zequanox)
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18. Questions?
Sarahann M. Rackl, Ph.D., P.E.
Marrone Bio Innovations
srackl@marronebio.com
(530) 750–2800
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