Toxicity of Arsenic to Fish - Society of Toxicology

Toxicity of Arsenic to Fish:
Role of Exposure Route, Fish
Species, and Chemical Form
Russell Erickson, David Mount, Terry Highland,
J. Russell Hockett, Dale Hoff, Correne Jenson, Tylor Lahren
U.S. Environmental Protection Agency
Office of Research and Development
Mid-Continent Ecology Division, Duluth, MN
Northland Society of Toxicology Meeting
St. Paul, MN
October 22, 1012
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Background
Clark
Fork
River
Ecological
Risk
Assessment
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Background
• Observation of reduced growth in fish fed diets prepared
from (a) invertebrates collected from Clark Fork River and
(b) invertebrates reared in Clark Fork River sediments.
• Laboratory dietary exposures of fish to individual
metal/metalloids indicated arsenic was likely the primary
cause for these growth effects.
• Further addressing this issue requires consideration:
The effects of the chemical speciation of arsenic
Combined effects of water and diet exposure
Relative sensitivity of different fish species
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Methods
• 28-60 day laboratory exposures of juvenile fish to arsenic in
diet and/or water; survival and growth monitored.
Swim-up rainbow trout (100-300 mg wwt)
30-d old fathead minnows (ca 100 mg wwt)
• Arsenic-contaminated diet provided as:
Live oligochaetes (separately exposed to arsenic
in water for 7-21 days, flow-through)
Pellets (arsenic solutions added to commercial
fish food, extruded into pellets, dried)
• Arsenic species used in exposures included arsenate (AsV),
arsenite (AsIII), monomethylarsonate (MMA), dimethylarsinate
(DMA), and arsenobetaine (AsB)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Arsenate: O=As(OH) 2 O -
-
Arsenite: As(OH) 2 O
Monomethylarsonate: O=As(CH 3 )(OH)O -
Dimethylarsinate: O=As(CH 3 ) 2 O -
Arsenobetaine: As(CH 3 3) 3 CH 2 COOH
Trimethylarsineoxide: O=As(CH 3 ) 3
Arsenocholine: As(CH 3 ) 3 CH 2 CH 2 OH
Arsenoylribosides: O=As(CH 3 ) 2 Sugar-R
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Live Diet Exposed to Arsenate or Arsenite
% Growth Re elative to C ontrol
4
100 3 12
3
3
3
80
60
Live Diet Exposed to Arsenate
40
Live Diet Exposed to Arsenite
1
Experiment Number
3
2
4
1
3
2
4
1
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Live Diet Exposed to Arsenate or Arsenite
% Growth Re elative to C ontrol
4
100 3 12
3
3
3
80
60
Live Diet Exposed to Arsenate
40
Live Diet Exposed to Arsenite
1
Experiment Number
3
2
4
1
3
2
4
1
Percen ntage Species in Live Diet
100
80
60
40
20
0
Arsenite Exposure
AsIII
AsV
MMA
DMA
AsB
AsX
Arsenate Exposure
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Live Diet Exposed to Arsenate or Arsenite
% Growth Re elative to C ontrol
100 43
12
80
60
3
3
3
Live Diet Exposed to Arsenate
40
Live Diet Exposed to Arsenite
1
Experiment Number
3
2
4
1
3
2
4
1
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Live Diet Exposed to Arsenate or Arsenite
Growth Re elative to Control
100
80
60
40
%
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Live Diet Exposed to MMA, DMA, AsB
% Growth Re elative to C ontrol
100
80
60
40
Live Diet Exposed to MMA
Live Diet Exposed to DMA
Live Diet Exposed to AsB
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Live Diet Exposed to MMA, DMA, AsB
% Growth Re elative to C ontrol
100
80
60
40
20
Live Diet Exposed to MMA
Live Diet Exposed to DMA
Live Diet Exposed to AsB
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Pe ercentage Species in Live Diet
100
80
60
40
20
0
MMA DMA AsB
Food Exposure
AsIII
AsV
MMA
DMA
AsB
AsX
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Pellet or Live Diet Contaminated with DMA, MMA, AsB, AsIII
% Growth Re elative to C ontrol
100
80
60
40
Pellet Diet Dosed with AsIII
Pellet Diet Dosed with MMA
Pellet Diet Dosed with DMA
Live Diet Exposed to MMA
Live Diet Exposed to DMA
Live Diet Exposed to AsB
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Pellet or Live Diet Contaminated with DMA, MMA, AsB, AsIII
% Growth Re elative to C ontrol
100
80
60
40
20
Pellet Diet Dosed with AsIII
Pellet Diet Dosed with MMA
Pellet Diet Dosed with DMA
Live Diet Exposed to MMA
Live Diet Exposed to DMA
Live Diet Exposed to AsB
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Pe ercentage Species in Pellet Diet
100
80
60
40
20
0
MMA DMA AsIII
Food Dosage
AsIII
AsV
MMA
DMA
AsB
AsX
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Pellet Dosed with DMA, MMA, AsIII, Live Diet Exposed to AsV
% Growth Re elative to Co ontrol
100
80
60
40
Live Diet Relationship
Pellet Diet AsIII
Pellet Diet MMA
Pellet Diet DMA
20
0.5 1 2 5 10 20 50 100
Arsenic in Fish (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Rainbow Trout in Clean Water
Pellet Dosed with DMA, MMA, AsIII, Live Diet Exposed to AsV
% Growth Re elative to C ontrol
100
80
60
40
20
Live Diet Relationship
Pellet Diet AsIII
Pellet Diet MMA
Pellet Diet DMA
0.5 1 2 5 10 20 50 100
Arsenic in Fish (μg/g dwt)
ish Tissue
rcentage Species in Fi
Per
100
80
60
40
20
0
MMA DMA AsIII
Food Dosage
AsIII
AsV
MMA
DMA
AsB
AsX
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Chemical Form
Summary/Conclusions (Dietborne Toxicity)
Arsenic speciation in diet matters and is strongly
reflected in accumulated species in fish
Inorganic arsenic more toxic than
common organic forms
Due to reducing environment, exposing diet organism
to AsIII or AsV does not affect dietary toxicity to fish
Pellet and live diets containing similar
arsenic species produce similar effects
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Rainbow Trout Exposed via Diet Only or Water Only
Live Diet Exposed to Various Levels of Arsenate
Growth Re elative to C ontrol
100
80
60
40
20
Fish Exposed to As
in Water, not Diet
Fish Exposed to As
in Diet, not Water
%
0
0.1 0.2 0.5 1 2 5 10 20
Arsenic in Water (mg As/L)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Rainbow Trout Exposed to Arsenate via Diet and Water
Live Diet Exposed to Various Levels of Arsenate
Growth Re elative to C ontrol
100
80
60
40
20
Control Diet
Diet Exposed
to 8 mg/L AsV
Diet Exposed
to 4 mg/L AsV
%
0
1 2 5 10
20
Fish Water Exposure (mg As/L)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Rainbow Trout Exposed to Arsenate via Diet and Water
Live Diet Exposed to Various Levels of Arsenate
4
Growth Re elative to C ontrol
100 0 0 0
80
60
40
0
20
8
Control Treatments
Control Diet
Diet Exposed to 4 mg/L AsV
Diet Exposed to 8 mg/L AsV
Nominal Water Conc mg As/L
0
8
0
16
0
8
0
16
8
16
%
0
0.5 1 2 5 10 20 50 100
Arsenic Accumulation in Fish (μg As/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Rainbow Trout Exposed to Arsenate via Diet and Water
Live Diet Exposed to Various Levels of Arsenate
100
80
% Survival
60
40 Control Treatments
Control Diet
Diet Exposed to ca. 4 mg As/L
20
Diet Exposed to ca. 8 mg As/L
0
0.1 0.2 0.5 1 2 5 10 20
Arsenic in Water (mg As/L)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Rainbow Trout Exposed to Arsenate via Diet and Water
Live Diet Exposed to Various Levels of Arsenate
100 0 0 0
0 0 8 00
8
8 8
4
80
16
% Survival
60
40
Control Treatments
Control Diet
Diet Exposed to 4 mg As/L
Diet Exposed to 8 mg As/L
16
16
20
0
Nominal Water Concentration
for Fish (mg/L AsV)
3232 32
0
0.5 1 2 5 10 20 50 100
Arsenic Accumulation in Fish (μg As/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Exposure Route
Summary/Conclusions (for Rainbow Trout)
For the same environmental concentration, diet route more
important for growth effects than water route.
Growth effects strongly correlated to accumulation in fish;
additive relationship for diet and water routes.
For the same environmental concentration, diet route
less important for lethal effects than water route.
Lethal effects strongly correlated to water concentrations,
and poorly related to accumulation in fish
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Fathead Minnow in Clean Water
Live Diet Exposed to Arsenate or Arsenite
Growth Re elative to C ontrol
100
80
60 Live Diet Exposed to AsV
Live Diet Exposed to AsIII
40
1
Experiment Number
1 1 1
2
2
2
%
20
1 2 5 10 20 50 100 200 500
Arsenic in Diet (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Fathead Minnow in Clean Water
Live Diet Exposed to Arsenate or Arsenite
ontrol
Growth Re elative to C
100
80
60 RBT Relationship
FHM, Live Diet Exposed to AsV
40
FHM, Live Diet Exposed to AsIII
%
20
1 2 5 10
20 50
Arsenic in Fish (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Fathead Minnow in Clean Water
Live Diet Exposed to Arsenate or Arsenite
Growth Re elative to C ontrol
100
80
60 RBT Relationship
FHM, Live Diet Exposed to AsV
40
FHM, Live Diet Exposed to AsIII
Percen ntage Species in Live Diet
100
80
60
40
20
0
FHM - AsIII
AsIII
AsV
MMA
DMA
AsB
AsX
RBT - AsIII
%
20
1 2 5 10
20 50
Arsenic in Fish (μg/g dwt)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Fathead Minnow with Water Exposure to Arsenate
Live Diet Exposed to Various Levels of Arsenate
100
80
Growth
%
60
40
20
Rainbow Trout Relationship
Control Diet
Diet Exposed to 2 mg/L AsV
Diet Exposed to 4 mg/L AsV
Diet Exposed to 8 mg/L AsV
0
0.2 0.5 1 2 5 10 20
Arsenic in Water (mg As/L)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Fathead Minnow with Water Exposure to Arsenate
Live Diet Exposed to Various Levels of Arsenate
Growth
%
100 0
0 0 0
80
Rainbow Trout
Relationship
Control Diet
60
Diet Level 1
Diet Level 2
Diet Level 3
40 Diet Level 4
Diet Level 5
Diet Level 6
20
0 Nominal Water
Concentration
0
0
0
44
4 4
8
8
8
8
0
0.5 1 2 5 10 20 50 100
Arsenic in Fish (mg As/L)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Fathead Minnow with Water Exposure to Arsenate
Live Diet Exposed to Various Levels of Arsenate
100
80
Survival
%
60
40
20
Rainbow Trout Relationship
Control Diet
Diet Expose to 2 mg/L AsV
Diet Exposed to 4 mg/L AsV
Diet Exposed to 8 mg/L AsV
0
0.2 0.5 1 2 5 10 20
Arsenic in Water (mg As/L)
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division

Role of Fish Species
Summary/Conclusions (Inorganic Arsenic)
FHM accumulates less total arsenic from diet than RBT,
and converts more to organic forms.
For growth, FHM less sensitive than RBT based on diet
concentration, but similar based on total accumulation.
FHM accumulates more arsenic from water than RBT, but
this does not contribute additively to growth effects.
For mortality, FHM more sensitive than RBT based on
water concentration, but not based on accumulation in fish.
Office of Research and Development
National Health and Ecological Effects Research Laboratory, Mid-Continent Ecology Division