Efficacy of Formulated Pf-CL145A
(Zequanox®) for Zebra Mussel Control in
Simulated Open Water Treatments
35th Annual Wisconsin Lakes Partnership
Convention April 9-11, 2013, Green Bay, WI
J.A. Luoma1, T.J. Severson1, K.L. Weber1, J.K. Wise1,
J.P. Motquin2*, D.A. Mayer3, and N.W. Olson4
1Upper
Midwest Environmental Sciences Center, La Crosse, Wisconsin 54603
2Shawano County Land Conservation Division, Shawano, Wisconsin 54166
3New York State Museum Field Research Laboratory, Cambridge, NY
4MN DNR, Ecological and Water Resources, Fergus Falls, MN
*Presenting author
U.S. Department of the Interior
U.S. Geological Survey
Partnership
Genoa
National
Fish
Hatchery
Shawano
County Land
Conservation
Division
Discovery of Pf-CL145A
Common soil bacterium
Targets digestive gland
Zequanox®
 Registered with USEPA
Normal tubule
 Defined discharge
 Tolerance exempt
 50% w/w Pf-CL145A
 Proprietary killed-cell
formulation
Necrotic tubule
Photos courtesy of New York State Museum
UMESC Grant Objectives
Non-target animal safety
 Unionid mussels
 Glochidia
 Juveniles
 Sub-adults
 Young-of-year fish
Treatment efficacy
 Simulated field trials
 In-Lake enclosures
Simulated Open Water Treatments
Evaluate efficacy of
formulated Pf-CL145A for
controlling zebra mussels
Treatment method
development
 Duration
 Concentration
 Application
Mobile Wet Laboratory
 Self-contained unit
 25 gpm pump with VFD
 Automatic backwash
filtration system
 Multiple effluent
management options
 Modular tank system
Test Tanks
Pump system
Removal of Treated
Water
Study Parameters
 Locations
 Lake Carlos (MN)
 Shawano Lake (WI)
 Application Techniques
 Whole Water Column
 Injection
 Pf-CL145A Treatments
 Control
 50 mg/L
 100 mg/L
Test Animal Collection
Collecting Zebra Mussels
Seeding
Substrate
Stack
Seeded
Substrate
Stacks
into
Cages
Distribution of Test Animals
 One substrate per bag
 > 50 attached ZM/tray  Empty shells removed
 Closed with zip‐ties
 Bags randomly
assigned exposure
chamber, tagged and
suspended at bottom of
tanks
Prepared
Bag
Suspending
bags
Dosing and Exposure
 Dosing
 0, 50, 100 mg/L active
 Triplicate tanks
 Whole Water Column Dosing
Whole
Water
Column
Dosing
 9 bags per tank
 6, 9, and 12 hour exposures
 Injection Dosing
 3 bags per tank
 12 hour exposure only
Injection
Dosing
Lake Water Chemistry
Dissolved
Temperature Hardness Alkalinity Conductivity
Oxygen
°C
mg/L
mg/L
µS/cm
mg/L
(STD)
(STD)
(STD)
(STD)
(STD)
Treatment
Location/type
pH Range
Lake Carlos W
8.60 – 8.65
8.76
(0.02)
22.2
(0.05)
177
(1.1)
163
(0.55)
395
(1.5)
Lake Carlos I
8.69 – 8.70
8.41
(0.02)
21.2
(0.03)
177
(1.0)
164
(0.41)
363
(3.3)
Shawano Lake W 9.31 – 9.34
7.36
(0.02)
22.6
(0.03)
118
(0.8)
105
(0.52)
248
(1.9)
Shawano Lake I 9.10 – 9.14
7.43
(0.02)
19.5
(0.00)
125
(1.1)
112
(0.55)
231
(2.6)
Pf-CL145A Concentration (mg/L)
Spectrophotometric Data
Lake Carlos
Whole Water Column
120
100
100
80
80
60
60
40
40
20
20
0
0
1
3
6
9
Shawano Lake
Whole Water Column
120
12
1
3
6
9
12
Time (hours)
Target Conc. 50 mg/L
Actual Conc. 50 mg/L
Target Conc. 100 mg/L
Actual Conc. 100 mg/L
Pf-CL145A Concentration (mg/L)
Spectrophotometric Data
Lake Carlos – Injection
Suspended and Bottom Sampling
300
300
250
250
200
200
150
150
100
100
50
50
0
0
1
3
6
9
Shawano Lake – Injection
Suspended Sampling
12
1
3
6
9
12
Time (hours)
Target Conc. 50 mg/L
Actual Conc. Suspended 50 mg/L
Actual Conc. Bottom 50 mg/L
Target Conc. 100 mg/L
Actual Conc. Suspended 100 mg/L
Actual Conc. Bottom 100 mg/L
Pf-CL145A Injection
Lake Carlos
Shawano Lake
Post-Dosing
 Bags removed and rinsed
 Placed in holding cages
 Held in lake for ~ 30 days
 Assessed Survival
Survival
Assessment
Holding
Cage
Whole Water Column Mortality
20
0
95.3
98.0
92.8
97.3
87.4
89.7
3.8
20
2.7
40
3.1
40
x̅ length =
18.6 mm
4.5
60
4.4
x̅ length =
11.4 mm
2.2
% Mortality
100
80
80
60
99.4
Shawano Lake
97.9
99.5
99.4
100
98.9
98.7
Lake Carlos
0
0
50
100
0
50
100
Pf-CL145A Concentration (mg/L)
6h
9h
12h
Injection Mortality
60
82.0
97.1
81.9
Lake Carlos
Shawano Lake
x̅ length
Carlos = 11.8 mm
Shawano = 18.7 mm
40
0
0
significant
difference in
LC 12-h
treatment type
WT vs. Inj.
P < 0.0001
No significant
difference in
SL 12-h
treatment type
WT vs. Inj.
P = 0.1686
4.5
20
2.7
% Mortality
80
99.1
100
50
100
Pf-CL145A Concentration (mg/L)
2013 Planned Research
 Sub-adult native mussels will be exposed
to Zequanox using procedures similar to
those used in 2012 ZM testing
 Limited scope, in-lake application within
enclosures at Keyes Lake, WI to native
mussels with attached zebra mussels
 LCCMR project
 Toxicity to aquatic insects
 Fathead minnow reproduction
 Limited in-lake applications in Lake
Minnetonka
QUESTIONS?
U.S. Department of the Interior
U.S. Geological Survey