Impacts of Selected Cumulative Pesticide Exposure/Risk Projects

Crofton,K.M.1, DeVito,M.J.1, Herr,D.W.1, Hughes,M.F.1, Lowitt,A.2, Baetcke,K.2, Moser,V.C.1, Padilla,S.1, Setzer,R.W.1, Gennings,C.4, Tornero,R.3
1
NHEERL, 3NERL, Office of Research and Development, and 2Office of Pesticide Programs, U. S. Environmental Protection Agency,
4
Virginia Commonwealth University, Richmond, Virginia

Carbamate Cumulative Risk Project Pyrethroid Cumulative Risk Project Results/Conclusions

Carbamates:
Science Questions •The carbamate studies are designed to: Obtain Free
PWG Samples of
In Vitro Human
“metabolism” Goal 2: Exposure to Dose to Response Assessment • All carbamates produce dose- and time-dependent
•Determine if the interaction of these seven pesticides in a mixture is dose-additive following acute exposure. OPP/FDA inhibition of cholinesterase and decreased motor
o What is the risk of exposure to mixtures of 11 Pyrethroids NERL
•Compare all cholinesterase assays using two different methods, a spectrophotometric method and a radiometric method. Sampling
Food
• Develop PBPK model for individual pyrethroids activity.
carbamate or pyrethroid insecticides? Survey Data
•Use motor activity as an indicator of neurological and behavioral impact of the pesticides. In Vitro Rat • Develop analytical procedures • The experimental mixture data will provide critical
•Correlate the behavioral and neurochemical endpoints for a better understanding of the biological effect of various levels of cholinesterase
Develop “metabolism” Generate
• Develop, parameterize and validate PBPK model data regarding potential interactions.
o What are appropriate exposure paradigms, and Analytical Individual
inhibition. Methods In Vivo Rat PBPK Models Generate • In vitro metabolism and dermal absorption of individual pyrethroids • Follow up with PBPK models allow integration of
what statistical models can be used to analyze
•Approach: Tissue Analyses “Relevant”
• Scale to human exposure biological data over a wide range of potential
these mixture data? Mixtures
•First, collect data on the individual carbamates. Adjust RPs • Limited in vivo studies to test PBPK model exposure scenarios.
•Time-course of cholinesterase inhibition in both the brain and red blood cells. Collect by TTcon • Develop PBPK model for mixtures of pyrethroids
•Dose-response assessment using brain and red blood cell cholinesterase inhibition as well as motor activity.
Effects & Firing Rate Pyrethroids:
Data
Generate • All pyrethroids produced dose-dependent
Research Goals •The mixture studies will be conducted using a fixed-ratio ray design. MA Dose
Response Generate Predictive
Scaling rat PBPK model to humans using depression in motor activity; relative potencies
•This allows for testing along a range of mixture doses wherein the proportion of pesticides within the mixture is the same. Relative Mixture Models

•One proportion tested will be based on projected environmental exposures from food, water, and air. ASR Dose Potencies in vitro and in vivo metabolism data ranged from 0.5 (λ-cyhalothrin) to 70 (resmethrin).
The FQPA requires consideration of cumulative Response • These data suggest a common endpoint for use in
exposures in determining the risk from exposure to •Another proportion tested will be based on the relative potency of the pesticides
•We will also measure carbamate levels in tissues and work with PBPK modelers in NHEERL and NERL to develop a model for cumulative
In Vitro
Firing Rates
Test Models
RAT IN VITRO RAT IN VIVO
In Vitro Clearance of cumulative risk calculations.
pesticides with a common mode of action. The With Relevant
• In vitro clearance data improves PBPK rat model.
exposure to carbamate pesticides. This approach has important utility given the relative short biological half-lives of the chemicals. Such PBPK
Rat/Human Mixtures in Rats Pyrethroids
current default for determining the cumulative risk to
pesticide mixtures assumes dose-additivity of models are able to simulate intermittent and repeated exposures of humans to carbamate pesticides. Elimination of Deltamethrin
Impact and Outcomes

L o g [D e lt a ] ( p m o le s /m L )
from Liver Microsomes
RAT PBPK MODEL
chemicals with a common mode-of-action. Previous Inform
Extrapolate
5
Cumulative Risk 0.1 M

efforts to determine cumulative risk of pesticides Assessments Model to Humans 4 0.5 M

1 M

with a common mode of action have been hampered

3

120 Propoxur
HUMAN PBPK MODEL 2 Impact and Outcomes
by a number of uncertainties. These obstacles Table 1. Carbamate pesticides, rat oral LD50 Dose-response Brain ChE
1
• This work will generate data and models for use in
include a lack of neurotoxicology mixture studies, assessment for motor
100 Horizontal activity 0

determining the cumulative risk of carbamates and

and dosages used. Vertical activity 0.0 2.5 5.0 7.5 10.0 12.5

and the unavailability of efficient statistical models

Time (min)

Goal 1: Neurotoxicity Studies

Percent control
80
activity and HUMAN IN VITRO HUMAN IN VIVO
pyrethroids.
with which to analyze mixtures data. 60
• These products will allow the Agency to conduct
Carbamate Rat oral Doses used** cholinesterase activity
The current effort has been developed based on state-of-the-art cumulative assessments for
LD50* for two carbamates.
40
• Dose response assessment for behavioral toxicity of pyrethroids Generic PBPK model
past work by NHEERL scientists in evaluating LC/MS of Deltamethrin carbamates and pyrethroids.
environmentally relevant mixtures of PHAHs, Methomyl 14-20 mg/kg 0.1-2.5 mg/kg For these pesticides, the
20
• Acute - Individual chemicals structure for pyrethroids
0
• Develop relative potency factors (administered dose vs. tissue dose) aerosol (broadcast, fogger)
• Uncertainties in the cumulative risk assessments
organophosphates (OP), and OP-carbamate mixtures. Methiocarb 10 mg/kg 0.5-25 mg/kg magnitude of activity 03 0
0.0. 1. 3.
0
10
. 0
20
.0
for these two economically important classes of
Two ongoing efforts aim to decrease the
Dose (mg/kg) • Mixtures - Assess assumption of additivity (Gennings et al, 2004) Lung

Oxamyl 2.5 mg/kg 0.07-1.5 mg/kg decreases is greater Venous side Arterial side pesticides will be reduced.
uncertainties in the cumulative risk assessments for than the cholinesterase 120
Methiocarb
Carbaryl 230 mg/kg 3-50 mg/kg Brain
carbamate and pyrethroid pesticides. Brain ChE

inhibition measured in 100 Horizontal activity

metabolism
Carbofuran 5-7 mg/kg 0.1-1.5 mg/kg Vertical activity
Pyrethroid Dose-effect Relative Potencies for Pyrethroids metabolism
the brain, illustrating

Percent control
Future Directions
80 Viscera

Formetanate 21 mg/kg 0.1-5 mg/kg Functions

the importance of
Fat
60
Specific aims: iv dosing
Propoxur 69 mg/kg 0.3-20 mg/kg 120

Motor Activity, % Control (mean ± SE)

dermal
1) To test the hypothesis of additivity. evaluating both 40 ED30* Relative
Pyrethroid Type (mg/kg) Potency*
100 Skin

2) Test the effects of mixtures based on * taken from MSDS endpoints. 20 Muscle
Carbamates
** no lethality occurred in any dose-response study 80
Deltamethrin II 3.0 1.0 dietary, hand to mouth, o Conduct fixed-ray design mixture studies.
environmentally relevant exposures (relative 0
05 0 0 .0 .0 object to mouth Liver

doses based on known human exposures).

0. 0. 2. 5. 12
Dose (mg/kg)
25 60
o Collect tissue level data for selected carbamates for
Esfenvalerate II 1.5 0.5 Volume of dist'n Excretion

3) Integrate the experimental neurotoxicology

40 CYPERMETHRIN
FENPROPATHRIN
DELTAMETHRIN (index chemical)
metabolism
use in kinetic modeling.
data with PBPK models and in vitro 20
CYFLUTHRIN
ESFENVALERATE
-CYHALOTHRIN
λ-Cyhalothrin II 1.4 0.5
Stomach Gut parent, metabolites in feces
o Complete PBPK models for individual chemicals as
mechanistic data. Comparison of the Brain Dose Response Comparison of the Brain Dose Response
0 well as mixture.
Radiometric Method
C V 0.01 0.03 0.1 0.3 1 3 10 30 100 300 β-Cyfluthrin II 2.8 0.9 Adjustments to PBPK Modeling for Deltamethrin in Rat*
Spectrophotometric Method Dose (mg/kg)

Pyrethroids
140
120
130
Cypermethrin II 12.0 4.0
110
o Complete PBPK models for single chemicals.
120

Motor Activity, % Control (mean ± SE)

120
Fenpropathrin I / II 9.0 3.0 Anadon
Cholinesterase (% control)

100
Cholinesterase (% control)

o Adjustment of relative potencies by tissue dose

110 100
90
100 Model
80 90 80
Tefluthrin I 4.0 1.3 from the PBPK models, two of which have been
References 70

60
80
70
60
Bifenthrin I 4.5 1.5
completed.
o Develop PBPK model for mixtures.
Propoxur Propoxur
60
50 Carbaryl Carbaryl 40
RESMETHRIN
Oxamyl 50
o Assess dose additivity using tissue dose.
Oxamyl s-BIOALLETHRIN

o Gennings C, Carter WH Jr, Carney EW, Charles GD, 40

30
Methomyl
Methiocarb
40 Methomyl 20
TEFLUTHRIN
BIFENTHRIN
s-Bioallethrin I 105 35.0
Methiocarb
Flow o Extrapolate model(s) to humans.
PERMETHRIN

Gollapudi BB, Carchman RA. (2004) A novel In vitro

30
Carbofuran Carbofuran DELTAMETHRIN (index chemical)
20
Formetanate 20 Formetanate
0
Permethrin I 70 23.3 Limited
flexible approach for evaluating fixed ratio mixtures 10 10
C V 0.01 0.03 0.1 0.3 1 3 10 30 100 300
Parameters
Dose (mg/kg)

of full and partial agonists. Toxicol. Sci. 80:134-150. 0

0.01 0.1 1 10 100
0
0.01 0.1 1 10 100 Resmethrin I 210 70.0 All Chemicals
Dose (mg/kg) Dose (mg/kg) o Work with OPP and NERL staff to design mixtures
Dose-Response assessment of 7 Carbamate Insecticides using Brain Cholinesterase of pyrethroids or carbamates based on food residue
○ Gray AJ, Rickard J (1982) The toxicokinetics of Inhibition as an Endpoint: Brain cholinesterase was assessed using two different Flow Adjusted and household residue (non-food) exposures.
deltamethrin in rats after intravenous administration methods, the radiometric method (left panel) and the Ellman method (right panel). The Limited PC (-30x) o Future work will include testing a series of relevant
of a toxic dose. Pestic. Biochem. Physiol. 18:205- radiometric method is superior for analysis of tissues from carbamate treated animals mixtures, and analyzing the neurotoxicology data
215. because that method limits reactivation and is less variable. for additivity.

*Data of Gray and Rickard (1982)

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