Efficacy of hydrogen peroxide to control Saprolegniasis on cool- and warmwater fish eggs
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Transcript of Efficacy of hydrogen peroxide to control Saprolegniasis on cool- and warmwater fish eggs
Efficacy of hydrogen peroxide to control Saprolegniasis on cool-
and warmwater fish eggs
Hydrogen peroxide INAD 10-023
(Perox-AidTM)
Egg Efficacy Trial Participants
• Mark Drobish – Gavins Point NFH
• Lynn Lee – UMESC
• Andy Moore – Rathbun Research
• Clark Moen – Blue Dog SFH
• Tom Harder – Max McGraw
• Dennis Smith – Dundee SFH
• Jerry Hamilton – Blind Pony SFH
INAD #10-023
• Study CAP-00-FUNGUS– submitted to CVM 03 March 2000– Twenty-nine participating facilities
• Study CAP-00-PARASITES– submitted to CVM 18 April 2000– Twenty-one participating facilities
• Study CAP-00-BACTERIA– submitted to CVM 18 April 2000– Twenty-three participating facilities
INAD #10-023 Participant Locations
UMESC
Western Chemicals
Eka Chemicals, Inc
Protocol Methods
Identify Disease
“Control”
Station History
“Prevention”
Select therapy and treat• concentration• duration• frequency• repetition
Chemical Inventory
Monitor / Record Mortality
Adverse Reactions
Monitor / Record Mortality
Controls?
Hydrogen peroxide therapy - Fish
• Duration: 30 or 60 min• Concentration
– 50, 75, or 100 mg/L for 60 min – 100 or 150 mg/L for 30 min
• Repetition/Interval: once daily on consecutive or alternate days for up to 10 treatments.
Hydrogen peroxide therapy - Eggs
• Duration: 15 min• Concentration: 500 or 750 mg/L
– minimum incubator concentration is 500 mg/L for 15 min
• Repetition/Interval: Once daily on consecutive or alternate days through hatch
Egg Efficacy Trials
• Seven egg efficacy trials conducted– Blind Pony SFH, Jerry Hamilton, paddlefish– Blue Dog SFH, Clark Moen, walleye– Dundee SFH, Dennis Smith, smallmouth bass– Gavins Point NFH, Mark Drobish, walleye– Max McGraw, Tom Harder, walleye– Rathbun SFH, Alan Moore, channel catfish– UMESC, Lynn Lee, walleye
• Fungus identified in three trials
Saprolegnia parasitica
Zoospores in zoosporangium
(blue arrow)
Gemmae (red arrow)
Photo credit: Steve Carpenter, Ph.D., Abbey Lane Labs.
Blind Pony SFH - paddlefish
• Temperature 15 - 16°C• Four trials - one or two females per trial
– 500, 750, or 1000 mg/L– % hatch ranged from 14 - 56% (of fertile eggs)– 2 - 4 consecutive treatments; fungus in all treated jars– noted cessation of egg development after hydrogen
peroxide treatment– withholding treatments until 2 d post fertilization resulted in
basically no hatch– compared rolling w/o treatment to hydrogen peroxide
treatment w/o rolling• rolled eggs - 60% hatch treated eggs 24% hatch
Blind Pony - paddlefishSaprolegnia ferax
Oogonium with oospores showing cell wall pitting.
Oogonia and gemmae from sterile house fly culture.
Photo credit: Steve Carpenter, Ph.D., Abbey Lane Labs.
Max McGraw - walleye
0102030405060708090
100
Per
cen
t h
atch
(%
)
Control 500
Treatment
rep 1
rep 2
rep 3
~ 1 million eggs total
x = 72% x = 68%
Blue Dog SFH - walleye
0
20
40
60
80
100
% e
yed
Form
- 1
Form
- 2
Form
- 3
H202
- 1
H2O2
- 2
H2O2
- 3Lot 5
Lot 4
Lot 3
~1.5 million eggs total
Gavins Point NFH - walleye
~3.1 million eggs total
58.365.5
75.5
0
20
40
60
80
100
Mea
n P
erce
nt
Hat
ch
Control
500 mg/L
750 mg/L
Gavins Point NFH - walleye
Control 500 mg/L 750 mg/L
Photo credit: Mark Drobish, Gavins Point NFH
UMESC - walleye
48.653.1* 49.7
65.9
78.1*70.0
0102030405060708090
100
Mea
n %
Hat
ch
Green Eyed
Control
500 mg/L
750 mg/L
~2.7 million eggs total
UMESC - Walleye
Overflow
Overflow
Plexiglass dividers
Perox-Aid stock solutions
Stock solution delivery lines
Headbox
Airstone Egg Jar
Pump
Hydrogen peroxide stock solutions
Dundee SFH - smallmouth bass eggs
01020
3040
5060
7080
90100
Per
cen
t h
atch
(%
)
Control 300 500 750
Treatment
rep 1
rep 2
rep 3
~ 2500 eggs per jar
~30,000 eggs total
Dundee SFH - smallmouth bass eggs
0
20
40
60
80
100
120
140
fung
used
egg
s
day 1
day 2
Number of fungused eggs removed during incubation.
Dundee SFH - smallmouth bass fry
0
10
20
30
40
50
60
70
80
90
100
Pe
rce
nt
su
rviv
al (
%)
Control 300 500 750
Treatment
rep 1
rep 2
rep 3
~ 2500 fry per tank
~30,000 fry total
Rathbun SFH - channel catfish eggs
0
1000
20003000
4000
5000
6000
70008000
9000
Nu
mb
er
of
fry
Rathbun SFH - channel catfish control eggs
Photo credit: Andy Moore, Rathbun SFH
Rathbun SFH - channel catfish treated eggs
Photo credit: Andy Moore, Rathbun SFH
Conclusions
• A hydrogen peroxide treatment regimen of 500 mg/L administered for 15 min on consecutive days from incubation to hatch is effective in controlling fungus on walleye and channel catfish eggs.
• Hydrogen peroxide virtually eliminated fungal growth in both walleye and channel catfish eggs and increased the probability of walleye hatch in two of three controlled trials.