Toxic Effects of Pesticides Matthew T. Martin, Ph.D. National Center for Computational Toxicology...
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Transcript of Toxic Effects of Pesticides Matthew T. Martin, Ph.D. National Center for Computational Toxicology...
Toxic Effects of Pesticides
Matthew T. Martin, Ph.D.National Center for Computational Toxicology Office of Research & DevelopmentU.S. Environmental Protection AgencyRTP, NC 27711Email: [email protected]://www.epa.gov/ncct/
Biochemical and Molecular ToxicologyUNC ENVR/TOXC 442November 8th, 2011
Pesticides
• Prevent, destroy, repel or mitigate any pest ranging from insects, animals and weeds, to microorganisms such as fungi, molds, bacteria and viruses
• Fungicides, Rodenticides, Herbicides, Insecticides, Antimicrobials• Inert & Other Ingredients make up final pesticide formulation
• OPP (US) & PMRA (Canada) Regulate Pesticides
• FIFRA, FFDCA, FQPA – Covers Most of US Pesticide Legislation
• Why have a lecture on pesticide toxicology?
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OPP= USEPA Office of Pesticide ProgramsPMRA=Pest Management Regulation AgencyFIFRA =Federal Insecticide, Fungicide, Rodenticide ActFFDCA=Federal Food, Drug, and Cosmetic ActFQPA =Food Quality Protection Act
Pesticides
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• Toxicity data rich chemicals
• Potential for high human exposure
• Designed to be bioactive
• What makes a pesticide different from a commodity chemical?– Designed to be bioactive– Is any pesticide just a pesticide? (“biocide”)– Statutory authority to require toxicity tests– Indirect & direct application to food/crops
• What makes a pesticide different from a pharmaceutical?– No direct human exposure– Design intentions (destroy vs treat)… Are they really different?– Molecular target potencies & efficacy differences
Pesticide Mass
Pesticides (by the numbers)
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• About 8800 total pesticidal ingredients
• About 4200 active ingredients– 1800 conventional pesticides– 300 antimicrobial pesticides– 250 biopesticides– 400 food-use (direct or indirect
contact with the food supply)– Remaining are unsupported
• About 4600 inert or other ingredients
• ~200 Chemical class represented
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Pesticides in the News
August 23, 2009Debating How Much Weed Killer Is Safe in Your Water Glass By CHARLES DUHIGGFor decades, farmers, lawn care workers and professional green thumbs have relied on the popular weed killer atrazine to protect their crops, golf courses and manicured lawns. But atrazine often washes into water supplies and has become among the most common contaminants in American reservoirs and other sources of drinking water. Now, new research suggests that atrazine may be dangerous at lower concentrations than previously thought. Recent studies suggest that, even at concentrations meeting current federal standards, the chemical may be associated with birth defects, low birth weights and menstrual problems. Laboratory experiments suggest that when animals are exposed to brief doses of atrazine before birth, they may become more vulnerable to cancer later. An investigation by The New York Times has found that in some towns, atrazine concentrations in drinking water have spiked, sometimes for longer than a month. But the reports produced by local water systems for residents often fail to reflect those higher concentrations.
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Pesticides in the News
October 7, 2009Regulators Plan to Study Risks of Atrazine By CHARLES DUHIGGThe Environmental Protection Agency plans to conduct a new study about the potential health risks of atrazine, a widely used weedkiller that recent research suggests may be more dangerous to humans than previously thought.Atrazine — a herbicide often used on corn fields, golf courses and even lawns — has become one of the most common contaminants in American drinking water. For years, the E.P.A. has decided against acting on calls to ban the chemical from environmental activists and some scientists who argued that runoff was polluting ecosystems and harming animals. More recently, new studies have suggested that atrazine in drinking water is associated with birth defects, low birth weights and reproductive problems among humans, even at concentrations that meet current federal standards.The E.P.A. is expected to announce on Wednesday that it will conduct a new evaluation of the pesticide to assess any possible links between atrazine and cancer, as well as other health problems, such as premature births. The E.P.A. may determine that new restrictions are necessary.
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Pesticides in the Scientific Literature
Silent SpringPublished in 1962
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Pesticides in the Regulatory Process
0
1000
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1945
1950
1955
1960
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1970
1975
1980
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1995
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2005
PESTICIDE REGULATORY SUBMISSIONS BY YEAR
Major AmendmentTo FIFRA
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Resource for High Quality Pesticide Chemical Use/Class Annotation
http://www.alanwood.net/pesticides/
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Pesticide Regulation• Federal Insecticide, Fungicide, and Rodenticide Act
(FIFRA, 1947) administered by USDA– Major amendments in 1972 and 1988
• Federal Food, Drug, and Cosmetic Act (FFDCA, 1954) established pesticide tolerances on food– Delaney Clause, forbade the use of carcinogens as food
additives• Food Quality Protection Act (FQPA, 1996) reauthorized
FFIFRA provisions– Tolerances reassessed as part of re-registrations– single, health-based standard– aggregate risk from all routes of non-occupational exposure– evaluating endocrine effects– extra tenfold uncertainty factor for children/in utero
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Vulnerability of Children
Greater exposure• On a caloric consumption:body-weight ratio Children are
2.5x adults. Diet less varied (fruit and milk)• Hand to mouth activity• Skin surface area per body weight is double that of an
adult• Rate of respiration
Greater physiological susceptibility• Period of rapid development of nerve cells• Loss of organ function can be permanently imprinted• Absorption and elimination of pesticides• Metabolizing enzymes not fully developed
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Pesticide Testing- US EPAHarmonized Test Guidelines
810 - Product Performance Test Guidelines 830 - Product Properties Test Guidelines 835 - Fate, Transport and Transformation Test Guidelines 840 - Spray Drift Test Guidelines 850 - Ecological Effects Test Guidelines 860 - Residue Chemistry Test Guidelines 870 - Health Effects Test Guidelines 875 - Occupational and Residential Exposure Test Guidelines 880 - Biochemicals Test Guidelines 885 - Microbial Pesticide Test Guidelines 890 - Endocrine Distruptor Screening Program Test Guidelines
http://www.epa.gov/ocspp/pubs/frs/home/guidelin.htm
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http://www.epa.gov/opp00001/reregistration/status.htm
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What is the acute and chronic Point-of-Departure for pesticide toxicity?
Reference Dose (RfD) = NOAEL x UF
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ToxRefDB website: http://actor.epa.gov/toxrefdb/
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Chronic Rat & Mouse Endpoints
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Reproductive Toxicity Profiling
http://toxsci.oxfordjournals.org/cgi/reprint/kfp080
Systemic Toxicity&
Delayed SexualMaturation
DecreasedReproductivePerformance
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Pesticide Carcinogenicity• Roughly 50% of all conventional pesticides cause tumors
in rodents• Generally pesticides are non-genotoxic carcinogens
(screened out in development process)• Human relevance?
– Site and tissue specificity (liver tumor in rodent vs lymphoma inc in humans)
– Mechanistic relevance (peroxisome proliferators)– High dose vs real world exposure potential
122
68
37
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No Pathology
Proliferative Lesions
Pre-neoplastic Lesions
Neoplastic Lesions
248 Chemicals122 w/ No Liver Pathology126 w/ Liver Pathology
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Chemicals Evaluated for Carcinogenic Potential by US EPA
http://www.epa.gov/pesticides/carlist/
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Is a pesticide an endocrine disruptor?
• Estrogenic• (anti) Androgenic• Thyrotoxic• Other
Panzica et al 2005
Endocrine Disruptor Screening Program
• The Food Quality Protection Act (FQPA) of 1996 and subsequent amendments to the Federal Food, Drug, and Cosmetic Act (FFDCA) and Safe Drinking Water Act (SDWA) required EPA to “develop a screening program, using appropriate validated test systems and other scientifically relevant information, to determine whether certain substances may have an effect in humans that is similar to an effect produced by a naturally occurring estrogen, or other such endocrine effect as the Administrator may designate.”
• The first phase of EDSP assays are designated Tier 1 tests with a purpose of identifying chemicals that exhibit potential to interact with endocrine pathways or mechanisms (i.e. the estrogen, androgen, and/or thyroid hormone systems) and ultimately determine which chemicals should undergo more definitive in vivo testing (i.e., Tier 2).
First test orders have been issued: http://www.epa.gov/endo/pubs/regaspects/testorders.htm
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http://www.epa.gov/endo/pubs/regaspects/testorders.htm
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Reproductive and
Endocrine Organ Toxicity
Endpoints from
ToxRefDB
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Interpretation of In Vitro Assay Results is Challenging
ERa_TRANS ERE_CIS
Concentration (µM)
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Importance of Biotransformation
ERa radioligand binding assay
ERa cellular (HEK293) transactivation assay
Parent/Metabolite
-3 -2 -1 0 1 2-20
0
20
40
60
80
100
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Methoxychlor (human ER)
HPTE (human ER)Methoxychlor (bovine ER)
HPTE (bovine ER)
BottomTopLogIC50HillSlopeIC50
HPTE (hER)2.448= 100.0-1.349-0.91970.04476
HPTE (bER)1.725= 100.0-1.697-0.98770.02009
Conc (log M)
% o
f In
hib
itio
n
Parent/Metabolite
-3 -2 -1 0 1 2-20
0
20
40
60
80
100
BottomTopLogEC50HillSlopeEC50
Methoxychlor0.301741.860.57145.4663.727
HPTE-3.91235.70-0.71312.2580.1936
Methoxychlor
HPTE
Conc (log M)
% o
f C
on
tro
l
Pesticidal MOA vs. Toxicological MOA
27http://www.irac-online.org/wp-content/uploads/2009/09/MoA_Classification.pdf
*Methoxychlor was intended to be a replacement for DDT (“Silent Spring”)
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EnvironmentalChemicals
Molecularresponse
Cellularresponse
Tissueresponse
Cell fate
Proliferation
Death Apoptosis Necrosis
AdverseOutcome
Hyperplasia
Tumor
Cancer
Chemicals
Pesticides Conazoles Pyrethroids
Toxics DE-71 PCBs Phthalates PFOA/PFOS
NR-sig Gene-reg. Transcription
CARPXRPPARa
cis-reg.trans-reg.
Xen. Met.Phase I
Phase II
Phase III
Molecular Response (Early)
NR activators stimulate intracellular processes that lead to hyperplasia
Chronic stimulation increases the risk of neoplasms
Focus On a Mode of Action …
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HISTORY OF DDT1,1,1-trichloro-2,2-bis-(p-chlorophenyl) ethane
• WWII – DDT was used by the allies to suppress a typhus epidemic in Naples
• 1943-1944 DDT was applied directly to the head of humans to control lice
• Success with DDT hastened the development of aldrin, dieldrin, endrin, chlordane, benzene hexachloride etc.
DDT was discovered to be an insecticide in 1939 by Paul Muller. He was a scientist working for Geigy, a Swiss firm that was focused on the chemical development of agricultural insecticides. Products with DDT entered the Swiss market in 1941. Seven years later, in 1948, Muller received the Nobel Prize for medicine and physiology in recognition for the lives DDT saved.
DDT• DDT can take more than 15 years to break down• Found in animals far from where they were it is used • Bio-accumulates in fish and marine mammals. Found concentrations in
these animals are many thousands of times higher than levels in water • DDT can be absorbed by some plants and by animals and humans who
eat those plants • DDT is fat-soluble and is stored in adipose tissues of humans and
animals
CURRENT STATUS:• No US registration, most uses cancelled in 1972, all uses by 1989• No US production, import, or export• DDE (metabolite of DDT) is regulated as a hazardous air pollutant
(Clear Air Act)• Priority toxic pollutant (Clean Water Act)
HUMAN EXPOSURE FROM:• Eating contaminated fish and shellfish • Eating imported food exposed to DDT • Infant exposed through breast milk • Eating products from crops grown in contaminated soil
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Insecticide advantages of DDT• Low volatility• Chemical stability• Lipid solubility• Slow rate of biotransformation and degradation
Disadvantages of DDT• Persistence in the environment• Bioconcentration• Biomagnification in food chain• Profound effects on wild life (“Silent Spring”)
Health Effects of DDT• Paresthesia of tongue, lips, and
face• Irritability, dizziness, vertigo,
tremor, and convulsions• Hypersusceptibility to external
stimuli (light, touch, and sound)
• Hypertrophy of hepatocytes• Hepatic tumors• No epidemiological evidence linking DDT
to carcinogenicity in humans• Low rate of absorption through the skin• Human health effects minor
34Klaassen, CD. CASARETT AND DOULL's Toxicology: The Basic Science of Poisons. McGraw-Hill 2001
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Sites of DDT poisoning
Klaassen, CD. CASARETT AND DOULL's Toxicology: The Basic Science of Poisons. McGraw-Hill 2001
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Neurotoxicity: Inhibition of choline esterase or action potential
• Organochlorine Insecticides
• Organophosphate Insecticides
• Carbamates• Pyrethroid
insecticides• Botanical
Insecticides
Klaassen, CD. CASARETT AND DOULL's Toxicology: The Basic Science of Poisons. McGraw-Hill 2001
• Most chemical insecticides act by poisoning the nervous system of the target organisms
• CNS of insects are highly developed and similar to that of the mammal
• Chemicals that act on the insect nervous system may have similar effects on higher forms of life
Stenersen, J. Chemical pesticides: Mode of Action and Toxicology. CRC Press 2004
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General Modes of Action
Pesticides acting on the axon (impulse transmission):
• Interference with transport of, Na+, K+, Ca2+, or Cl- ions
Pesticides acting on synaptic transmission:
• Inhibition of specific enzyme activities:GABA-ergic (inhibitory) synapsesCholinergic synapses
• Contribution to the release or persistence of chemical transmitters at nerve endings
Stenersen J, Chemical Pesticides Mode of Action and Toxicology, CRC Press 2004
39
Pyrethroid Insecticides
• Newest class of insecticides• New analogs will be (hopefully):
– More stable in light and air– Better persistence– Low mammalian toxicity
Soderlund et al. (2002)
Importance of Structure-Activity-Toxicity
Relationships
Soderlund et al. (2002)
41
Pyrethroid Use
• Household sprays• Flea preparations for pets• Plant sprays for home• Plant sprays for greenhouses
• Similar to DDT• Not highly toxic in animals• Toxic ingredients
– Chrysanthemic acid– Pyrethric acid
Pyrethroid Poisoning
Figure 1. Nine neonicotinoid insecticides and four nicotinoids. The neonicotinoids are nitromethylenes (C==CHNO2), nitroguanidines (C==NNO2), and cyanoamidines(C==NCN). Compounds with 6-chloro-3-pyridinylmethyl, 2-chloro-5-thiazolylmethyl, and 3-tetrahydro-furanmethyl moieties are referred to as chloropyridinyls (or chloronicotinyls),chlorothiazolyls (or thianicotinyls), and tefuryl, respectively. The nicotinoidsare naturally occurring [(−)-nicotine and (−)-epibatidine] and synthetics (ABT-594 and desnitroimidacloprid).
Tomizawa & Casida (2004)
Tomizawa & Casida (2004)