Parkinson's disease and residential exposure to maneb and paraquat from agricultural applications in the central valley of California.
ABSTRACT Evidence from animal and cell models suggests that pesticides cause a neurodegenerative process leading to Parkinson's disease (PD). Human data are insufficient to support this claim for any specific pesticide, largely because of challenges in exposure assessment. The authors developed and validated an exposure assessment tool based on geographic information systems that integrated information from California Pesticide Use Reports and land-use maps to estimate historical exposure to agricultural pesticides in the residential environment. In 1998-2007, the authors enrolled 368 incident PD cases and 341 population controls from the Central Valley of California in a case-control study. They generated estimates for maneb and paraquat exposures incurred between 1974 and 1999. Exposure to both pesticides within 500 m of the home increased PD risk by 75% (95% confidence interval (CI): 1.13, 2.73). Persons aged < or =60 years at the time of diagnosis were at much higher risk when exposed to either maneb or paraquat alone (odds ratio = 2.27, 95% CI: 0.91, 5.70) or to both pesticides in combination (odds ratio = 4.17, 95% CI: 1.15, 15.16) in 1974-1989. This study provides evidence that exposure to a combination of maneb and paraquat increases PD risk, particularly in younger subjects and/or when exposure occurs at younger ages.
- SourceAvailable from: Areski Chorfa[Show abstract] [Hide abstract]
ABSTRACT: The etiology of most human neurodegenerative disorders is believed to be multifactorial and consists of an interaction between environmental factors and genetic predisposition. Regarding Parkinson's disease (PD), the second most frequent neurodegenerative disease characterized by the aggregation of the alpha-synuclein (αS) protein, numerous epidemiological and experimental studies suggested a possible role of exposure to some pesticides. Whereas epidemiological studies largely failed to identify pesticides specifically involved in PD, it is of critical importance to set up in vitro toxicity studies of pesticides. We measured changes of αS levels following pesticide exposures of human cell lines in vitro, using either ELISA detection of endogenous αS or flow cytometry after overexpression using recombinant adenoviruses. We showed that three pesticides (paraquat, rotenone and maneb), which have frequently been associated with PD, produced a dose-dependent increase in cellular αS levels, but also of αS released into the culture medium. Examining an additional series of 20 pesticides from different families and chemical structures, we found that beside some insecticides, including an organophosphate and three pyrethroids, a majority of the 12 studied fungicides were also producing an αS accumulation, three of them (thiophanate-methyl, fenhexamid and cyprodinil) having similar or more pronounced effects than paraquat. A variety of pesticides can disrupt αS homeostasis in vitro; our data illustrate an experimental strategy that could help in the identification of chemicals that could be specifically involved in PD etiology.Archive für Toxikologie 10/2014; · 5.08 Impact Factor
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ABSTRACT: Treatment with rotenone, both in vitro and in vivo, is widely used to model dopamine neuron death in Parkinson's disease upon exposure to environmental neurotoxicants and pesticides. Mechanisms underlying rotenone neurotoxicity are still being defined. Our recent studies suggest that rotenone-induced dopamine neuron death involves microtubule destabilization, which leads to accumulation of cytosolic dopamine and consequently reactive oxygen species (ROS). Furthermore, the c-Jun N-terminal protein kinase (JNK) is required for rotenone-induced dopamine neuron death. Here we report that the neural specific JNK3 isoform of the JNKs, but not JNK1 or JNK2, is responsible for this neuron death in primary cultured dopamine neurons. Treatment with taxol, a microtubule stabilizing agent, attenuates rotenone-induced phosphorylation and presumably activation of JNK. This suggests that JNK is activated by microtubule destabilization upon rotenone exposure. Moreover, rotenone inhibits VMAT2 activity but not VMAT2 protein levels. Significantly, treatment with SP600125, a pharmacological inhibitor of JNKs, attenuates rotenone inhibition of VMAT2. Furthermore, decreased VMAT2 activity following in vitro incubation of recombinant JNK3 protein with purified mesencephalic synaptic vesicles suggests that JNK3 can inhibit VMAT2 activity. Together with our previous findings, these results suggest that rotenone induces dopamine neuron death through a series of sequential events including microtubule destabilization, JNK3 activation, VMAT2 inhibition, accumulation of cytosolic dopamine, and generation of ROS. Our data identify JNK3 as a novel regulator of VMAT2 activity. Copyright © 2014. Published by Elsevier Ireland Ltd.Toxicology 12/2014; · 3.75 Impact Factor
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ABSTRACT: Pesticides have been associated with Parkinson's disease (PD), and protective gloves and workplace hygiene can reduce pesticide exposure. We assessed whether use of gloves and workplace hygiene modified associations between pesticides and PD. The Farming and Movement Evaluation (FAME) study is a nested case–control study within the Agricultural Health Study. Use of protective gloves, other PPE, and hygiene practices were determined by questionnaire (69 cases and 237 controls were included). We considered interactions of gloves and hygiene with ever-use of pesticides for all pesticides with ≥ 5 exposed and unexposed cases and controls in each glove-use stratum (paraquat, permethrin, rotenone, and trifluralin). 61% of respondents consistently used protective gloves and 87% consistently used ≥ 2 hygiene practices. Protective glove use modified the associations of paraquat and permethrin with PD: neither pesticide was associated with PD among protective glove users, while both pesticides were associated with PD among non-users (paraquat OR 3.9 [95% CI 1.3, 11.7], interaction p = 0.15; permethrin OR 4.3 [95% CI 1.2, 15.6] interaction p = 0.05). Rotenone was associated with PD regardless of glove use. Trifluralin was associated with PD among participants who used < 2 hygiene practices (OR 5.5 [95% CI 1.1, 27.1]) but was not associated with PD among participants who used 2 or more practices (interaction p = 0.02). Although sample size was limited in the FAME study, protective glove use and hygiene practices appeared to be important modifiers of the association between pesticides and PD and may reduce risk of PD associated with certain pesticides.Environment International 11/2014; · 5.66 Impact Factor
American Journal of Epidemiology
ª The Author 2009. Published by the Johns Hopkins Bloomberg School of Public Health.
All rights reserved. For permissions, please e-mail: firstname.lastname@example.org.
Vol. 169, No. 8
Advance Access publication March 6, 2009
Parkinson’s Disease and Residential Exposure to Maneb and Paraquat From
Agricultural Applications in the Central Valley of California
Sadie Costello, Myles Cockburn, Jeff Bronstein, Xinbo Zhang, and Beate Ritz
Initially submitted September 12, 2008; accepted for publication January 6, 2009.
Evidence from animal and cell models suggests that pesticides cause a neurodegenerative process leading to
Parkinson’s disease (PD). Human data are insufficient to support this claim for any specific pesticide, largely
because of challenges in exposure assessment. The authors developed and validated an exposure assessment
tool based on geographic information systems that integrated information from California Pesticide Use Reports
and land-use maps to estimate historical exposure to agricultural pesticides in the residential environment. In
1998–2007, the authors enrolled 368 incident PD cases and 341 population controls from the Central Valley of
California in a case-control study. They generated estimates for maneb and paraquat exposures incurred between
1974 and 1999. Exposure to both pesticides within 500 m of the home increased PD risk by 75% (95% confidence
interval (CI): 1.13, 2.73). Persons aged ?60 years at the time of diagnosis were at much higher risk when exposed
to either maneb or paraquat alone (odds ratio ¼ 2.27, 95% CI: 0.91, 5.70) or to both pesticides in combination
(odds ratio ¼ 4.17, 95% CI: 1.15, 15.16) in 1974–1989. This study provides evidence that exposure to a combi-
nation of maneb and paraquat increases PD risk, particularly in younger subjects and/or when exposure occurs at
case-control studies; fungicides, industrial; geographic information systems; herbicides; maneb; paraquat;
Parkinson disease; pesticides
Abbreviations: CI, confidence interval; DDE, dichlorodiphenyldichloroethylene; GIS, geographic information system; MPPþ, toxic
metabolite of 1-methyl-4-phenylpyridinium; OR, odds ratio; PD, Parkinson’s disease; PLSS, Public Land Survey System; PUR,
Pesticide Use Reporting.
Parkinson’s disease (PD) has been reported to occur at
high rates among farmers and in rural populations, contrib-
uting to the hypothesis that agricultural pesticides might be
causal agents (1–4). Animal studies have linked certain pes-
ticides to Parkinsonism and dopaminergic cell death. The
pesticide rotenone can produce the behavioral and neuro-
pathologic features of PD in some rodent models through
chronic systemic inhibition of mitochondrial complex I (5,
6). Exposure to a combination of the fungicide maneb and
the herbicide paraquat in mice leads to increased substantia
nigra neuronal pathology (7), age-dependent motor degen-
eration, progressive reductions in dopamine metabolites and
turnover (8), and reduced tyrosine hydroxylase and dopa-
mine transporter immunoreactivity (9, 10).
Human evidence is insufficient to identify any particular
pesticide compound, including those implicated by animal
studies, as being responsible for causing PD (11). Method-
ological limitations have clouded the interpretation of most
epidemiologic studies exploring pesticide exposures and PD
in humans. Past studies have generally relied on self-reports
and recall of chemical usage, making them vulnerable to
information bias and differential recall bias (12).
Because pesticides applied from the air or ground may
drift from their intended treatment sites, with measurable
concentrations subsequently detected in the air, in plants,
and in animals up to several hundred meters from applica-
tion sites (13–15), accurate methods of estimating environ-
mental exposures in rural communities are sorely needed.
Correspondence to Dr. Sadie Costello, Department of Environmental Health Sciences,School of Public Health, University of California, Berkeley,
50 University Hall, #7360, Berkeley, CA 94720-7360 (e-mail: email@example.com).
919 Am J Epidemiol 2009;169:919–926
Geographic information system (GIS)-based methods of as-
sessing exposure to pesticides have become popular in re-
cent years and may prove an effective solution when
pesticide data exist. We developed and employed avalidated
GIS-based exposure assessment tool to estimate pesticide
exposure from applications to agricultural crops, relying
on California Pesticide Use Reporting (PUR) data, land-
use maps, and geocoded residential historical locations
(16). We investigated whether exposure to the pesticides
maneb and paraquat, alone and in combination, increased
the risk ofincident PD among residents of the CentralValley
of California, an area well-known for its intensive agricul-
ture and potential for pesticide exposure.
MATERIALS AND METHODS
All procedures described have been approved by the
University of California, Los Angeles, institutional review
board for human subjects, and informed consent was ob-
tained from all participants.
We used a population-based approach for recruiting cases
and controls from a largely agricultural population in Cali-
fornia. Details are provided elsewhere (17). Briefly, persons
with PD newly diagnosed between January 1998 and January
2007 who resided in 1 of 3 central California counties
(Fresno, Tulare, or Kern county) and had lived in California
for at least 5 years prior to diagnosis were recruited into our
study within 3 years of diagnosis. Altogether, 28 (90%) of
the 31 practicing local neurologists who provided care for
PD patients assisted in recruiting cases for this study. We
solicited collaboration from Kaiser Permanente Medical
California), and Visalia Medical Clinic (Visalia, California)
and from the Veterans Administration, PD support groups,
local newspapers, and local radio stations that broadcast
public service announcements.
Of the 1,167 PD cases who were initially invited, 604
were not eligible: For 397, the case’s diagnosis date fell
outside the 3-year range prior to contact, 51 denied having
received a PD diagnosis, 134 lived outside the tricounty
area, and 22 were too ill to participate. Of the 563 eligible
cases, 473 (84%) were examined by a University of Cali-
fornia, Los Angeles, movement disorder specialist at least
once and were confirmed to have clinically ‘‘probable’’ or
‘‘possible’’ PD; the remaining 90 potential cases could not
be examined or interviewed (54% withdrew, 32% were too
ill or died, and 14% moved out of the area prior to the
examination or did not honor a scheduled appointment).
We examined but excluded another 96 patients because they
had other causes of Parkinsonism. This left us with 377
cases; of these, 368 provided all information needed for
Controls aged 65 years or older were identified from
Medicare lists in 2001, but because of implementation of
the Health Insurance Portability and Accountability Act,
which prohibits the use of Medicare enrollees, 70% of our
controls were recruited from randomly selected tax assessor
residential units (parcels) in each of the 3 counties. We
mailed letters of invitation to a random selection of residen-
tial living units and also attempted to identify head-of-
household names and telephone numbers for these parcels,
using the services of marketing companies and Internet
We contacted 1,212 potential population controls by mail
and/or telephone for eligibility screening. Eligibility criteria
were: 1) not having PD, 2) being at least 35 years of age,
3) currently residing primarily in 1 of the 3 designated coun-
ties, and 4) having lived in California for at least 5 years
prior to the screening. Only 1 person per household was
allowed to enroll. Of the potential controls contacted, 457
were ineligible: 409 were too young, 44 were terminally ill,
and 4 resided primarily outside of the study area. Of the 755
eligible controls, 409 (54%) declined participation, were too
ill to honor an appointment, or moved out of the area prior to
interview; 346 (46%) were enrolled, and 341 provided all
information needed for analyses.
Assessment of environmental pesticide exposure
We conducted telephone interviews to obtain demo-
graphic and exposure information. Detailed residential his-
tory forms were mailed to subjects in advance of their
interview and were reviewed in person or over the phone.
We estimated pesticide exposures in the residential environ-
ment from applications to agricultural crops employing
a validated GIS-based system, which combined PUR data
and land-use maps (16, 18), to produce estimates of residen-
tial ambient pesticide applications within a set distance of
subjects’ homes. We recorded and geocoded lifetime resi-
dential histories and estimated ambient exposures for all
historical addresses at which participants had resided be-
tween 1974 and 1999, the period covered by the PUR data.
A technical discussion of our GIS-based approach is pro-
vided elsewhere (16); here we briefly summarize the data
sources and the exposure modeling process.
Residential addresses. Addresses were automatically
geocoded to TigerLine files (NAVTEQ (Chicago, Illinois),
unpublished data, 2006), and discrepancies were then man-
ually resolved in a multistep process similar to that de-
scribed by McElroy et al. (19). Resulting locations were
recorded, along with the relevant year range of residence,
so they could be matched to the appropriate year-specific
PUR and land-use data (below). For our GIS model, we
relied on addresses in Fresno, Kern, and Tulare counties
(the tricounty area) at which participants had resided be-
tween 1974 and 1999. Out of 9,568 total residential years
contributed by cases (26 years 3 368 cases), 7,593 years
(79%) were spent at addresses within the tricounty area as
compared with 6,757 (76%) of 8,866 years contributed by
controls (26 years 3 341 controls). We geocoded these
tricounty residential addresses for the period 1974–1999
with similar precision for cases and controls; that is, both
had spent 88% of their respective residential years at ad-
dresses we considered to have been mapped with high pre-
cision (i.e., at the level of a residential parcel, street
address, or street intersection rather than a zip code or city
920Costello et al.
Am J Epidemiol 2009;169:919–926
Pesticide use reporting. PUR data are recorded by the
California Department of Pesticide Regulation for any com-
mercial application of restricted-use pesticides (defined as
agents with harmful environmental or toxicologic effects
(20)) and, since 1990, for all commercial uses of pesticides
regardless of toxicologic profile. The location of each PUR
record is referenced to the Public Land Survey System
(PLSS), a nationwide grid that parcels land into sections
at varying resolutions. Each PUR record includes the name
of the pesticide’s active ingredient, the poundage applied,
the crop and acreage of the field, the application method,
and the date of application.
Land-use maps. Because the PUR records link an agri-
cultural pesticide application only to a whole PLSS grid
section, we added information from land-use maps to more
precisely locate the pesticide application, as described in
detail elsewhere (18). The California Department of Water
Resources periodically (every 7–10 years) performs county-
wide large-scale surveys of land use and crop cover, which
allowed us to identify the locations of specific crops within
each PLSS grid section. Digital maps from more recent
(1996–1999) surveys are available (21), and paper maps
were manually digitized for earlier periods (1977–1995).
The 1977 land-use survey was conducted closest in time
to 1974, when PUR data became available. We constructed
historical electronic maps of land use and crop type, and
using the PLSS grid section and the crop type reported in the
PUR record, we allocated pesticide applications to an agri-
cultural site to which we assigned a GIS-based location.
Deriving estimates of residential pesticide exposure. The
time-specific total exposure at each location, by pesticide,
was derived through summation of exposures over a fixed
500-m radius (suggested in previous literature (13, 15, 19))
around the home for the relevant years of residence. The
numbers of pounds of pesticide applied annually per acre
were summed for each residential buffer and weighted by
the proportion of treated acreage in each buffer, resulting in
pesticide application rates that could be averaged over spe-
cific calendar periods of each subject’s lifetime.
We estimated residential exposures to maneb and para-
quat, alone and in combination, for the following time win-
dows: 1) 1974–1999, 2) 1974–1989, and 3) 1990–1999, to
assess the possibility of an extensive induction period prior
to PD onset and the influence of age at exposure. We strat-
ified models by sex and age (?60 years, >60 years) and, in
additional sensitivity analyses, controlled for exposure to
some groups of pesticides suspected to increase PD risk.
We controlled for occupational exposure to pesticides
among subjects who had held jobs in the agricultural sector,
assigning them to categories of ‘‘likely exposed to pesti-
cides’’ when they reported pesticide handling and applica-
tions or fieldwork and ‘‘possibly exposed to pesticides’’
when they reported managerial, produce processing, and
other nonfield farm work; all other subjects were considered
‘‘not occupationally exposed to pesticides’’ (22). In some
models, we also adjusted for residential exposures to groups
of other pesticides that some studies have found to be linked
to dopaminergic cell damage or possibly PD (organochlo-
rines,organophosphates,and dithiocarbamates (23) and pro-
teasome inhibitors (24)).
We considered the following demographic variables as
potential confounders in all analyses: age (age at diagnosis
for cases and age at interview for controls), sex, race (white,
nonwhite), education (<12 years, 12 years, >12 years), and
cigarette smoking (current, former, never). We used SAS 9.1
(SAS Institute Inc., Cary, North Carolina) to perform un-
conditional logistic regression analyses.
Study participants were predominantly Caucasian, over
the age of 60, and without a family history of PD (Table 1).
Cases were slightly older than controls, were more often
male, and had completed fewer years of education. They
were also more likely to have been occupationally exposed
to pesticides and to be never or former smokers.
We did not find increased risks of PD among subjects
exposed to paraquat alone during the years 1974–1999
(Table 2). While the rarityof sole manebexposure (4subjects)
precluded any meaningful interpretation of the maneb-only
results, combined exposure to both maneb and paraquat in-
creased the risk of PD by 75% (odds ratio (OR) ¼ 1.75,
95% confidence interval (CI): 1.13, 2.73), an effect estimate
which was essentially unchanged after adjustment for
occupational pesticide exposure (OR ¼ 1.74, 95% CI:
When we examined 2 separate exposure time windows,
the years 1974–1989 and 1990–1999, the risk increase ob-
served for the whole period was found to be mainly attribut-
able to exposures incurred during the earlier window
(OR ¼ 2.14, 95% CI: 1.24, 3.68), while being exposed
during the later window did not seem to increase PD risk
(Table 2). Furthermore, for younger (?60 years) subjects,
exposure to both maneb and paraquat in both windows in-
creased PD risk as much as 4- to 6-fold (Table 3). Exposure
to either maneb or paraquat alone during 1974–1989 also
increased risk of PD in younger subjects (OR ¼ 2.27, 95%
CI: 0.91, 5.70). When we examined exposure windows
among our older subjects (>60 years), combined exposure
to both pesticides in the earlier window only (1974–1989)
was also associated with a 2-fold increase in PD risk
(OR ¼ 2.15, 95% CI: 1.15, 4.02), but no increase was found
for either the later window (1990–1999) or the combined
exposure periods (Table 3). Stratification by sex suggested
no differences in estimates between males and females.
In this population-based case-control study, agricultural
application of both maneb and paraquat within 500 m of
a residence during the period 1974–1999 greatly increased
the risk of developing PD, especially when exposure oc-
curred between 1974 and 1989 or when PD was diagnosed
at a younger age (?60 years). Exposure to both pesticides
during the earlier time window (1974–1989) also doubled
the risk for older cases. Associations were particularly
PD and Residential Maneb and Paraquat Exposure921
Am J Epidemiol 2009;169:919–926
strong for younger-onset patients (?60 years), who would
have been children, teenagers, and young adults during the
exposure period: Among those exposed in the earlier time
window, risk was increased more than 4-fold with exposure
to both pesticides and more than 2-fold with exposure to just
1 of the pesticides. Consistent with some theories regarding
the progression of PD pathology (25), these data suggest
that the critical window of exposure to toxicants may be
years before the onset of motor symptoms which lead to
Pesticide and herbicide exposures have previously been
implicated in idiopathic PD. Paraquat is structurally similar
to the toxic metabolite (MPPþ) of the 1-methyl-4-phenyl-
pyridinium ion (a metabolite of 1-methyl-4-phenyl-1,2,3,6-
tetrahydropyridine), an agent known to induce Parkinsonian
symptoms in humans that has been widely used to study
Characteristics, Central Valley of California, 1998–2008
Odds Ratio for Parkinson’s Disease According to Various Sociodemographic
(n 5 368)
(n 5 341)
No. or Mean% No. or Mean%
68.1 (34–88)67.6 (34–92)1.00 0.99, 1.02
Age group, years
51–60 47 135516
61–70 1113095 28
71–80 145 39121 35
Female sex 16144 165480.83 0.62, 1.12
First-degree relative with
55 1537111.440.93, 2.25
296 80279 821 Reference
72 2062 181.09 0.75, 1.60
Native American164 103
<12 6818 38111.15 0.69, 1.90
>1220054 239700.54 0.37, 0.77
Job exposure matrix
exposed to pesticides
232 63 240701Reference
exposed to pesticides
2672681.03 0.58, 1.83
exposed to pesticides
1103075221.52 1.08, 2.14
Cigarette smoking status
Never smoker 19553 146431Reference
Former smoker 15141161 470.70 0.52, 0.96
Current smoker226 34100.48 0.27, 0.86
0 19553 146 431Reference
9626 89260.81 0.56, 1.16
7721 106310.54 0.38, 0.78
aThe odds ratio was calculated for all nonwhites versus whites.
922 Costello et al.
Am J Epidemiol 2009;169:919–926
Parkinsonism in animal models (26). MPPþ is believed to
cause cell death by interfering with mitochondrial respira-
tion (27), because it concentrates in mitochondria and in-
hibits complex I of the electron transport chain (28). Many
lines of evidence point to possible mitochondrial dysfunc-
tion in PD. Several genes have been identified in familial
forms of PD that are linked to mitochondrial function
(PINK1 and DJ1), and in sporadic cases of PD, pathologic
free radical reactions that damage mitochondria and de-
crease electron transport activity have been described (29).
Impaired electron transport hampers adenosine triphosphate
production and leads to the diversion of electrons from their
normal electron transport recipients and, thus, further for-
mation of damaging free radicals (29).
Although paraquat is also used to induce Parkinsonism in
some animal models, the mechanism by which it produces
symptoms is not yet understood (30). Recent mammalian
and yeast-cell experiments suggest that mitochondria take
up paraquat actively across their membranes, where com-
plex I reduces it to the paraquat radical cation that subse-
quently produces mitochondria-damaging superoxide (31).
It has also been suggested that maneb may inhibit the ubiq-
uitin proteasome system, thereby damaging the dopaminer-
gic neuron (24, 32). Additionally, maneb has been linked to
Parkinsonism in mice also exposed to paraquat. In 3 recent
studies, investigators reported that only when mice were
exposed to a combination of the fungicide maneb and the
Residential Ambient Exposure to Maneb and/or Paraquat, Central
Valley of California, 1974–1999
Odds Ratio for Parkinson’s Disease According to
(n 5 368)
(n 5 341)
Missing data134 134
No exposure11531 126 371Reference
Paraquat only149 40 152451.01 0.71, 1.43
Maneb only31103.04 0.30, 30.86
8824 49 141.75 1.13, 2.73
Missing data 53 1452 15
No exposure 9325 113331Reference
14840 13740 1.250.85, 1.85
742039112.14 1.24, 3.68
Missing data 154 154
No exposure21558213 621Reference
113 3195280.96 0.64, 1.43
257185 0.930.45, 1.94
aOdds ratios were adjusted for age, sex, nonwhite race, education,
and smoking status. Results were mutually adjusted for exposure in
each time window.
Residential Ambient Exposure to Maneb and/or Paraquat, by Time
Window of Exposure and Age Group, Central Valley of California,
Odds Ratio for Parkinson’s Disease According to
1974–1999 Time Window
No exposure 18 2334391Reference
38 4842 481.77 0.84, 3.75
21 27785.07 1.75, 14.71
Missing data 11494
No exposure97 3492361Reference
114 39 111440.90 0.60, 1.34
1974–1989 Time Window
Missing data162020 23
364634 392.270.91, 5.70
14 18674.171.15, 15.16
Missing data371332 13
No exposure80 2886341Reference
60 2133132.151.15, 4.02
1990–1999 Time Window
No exposure43 5458671Reference
No exposure172 60 155 611Reference
aAge-stratified models with adjustment for sex, nonwhite race,
education, and smoking status. Results were mutually adjusted for
exposure in each time window.
PD and Residential Maneb and Paraquat Exposure923
Am J Epidemiol 2009;169:919–926
herbicide paraquat (paraquat þ maneb), not to either pesti-
cide alone, did they exhibit increased neuronal pathology
(7), age-dependent motor degeneration and progressive re-
ductions in dopamine metabolites and dopamine turnover
(8), and reduced tyrosine hydroxylase and dopamine trans-
porter immunoreactivity (9).
The fungicide maneb and the herbicide paraquat are both
used in the Central Valley of California and are often used
on the same crops, including potatoes, dry beans, and toma-
toes. The average amount of maneb applied near the homes
of these study subjects was relatively stable throughout both
time windows; however, annual paraquat exposure in-
creased during the later (1990–1999) time window. Persons
living near fields sprayed with maneb and paraquat may also
be exposed to a host of other agricultural chemicals. When
we controlled for the influence of other groups of pesticides
suspected a priori to be risk factors for PD in our study, the
odds ratios for combined maneb and paraquat exposure and
PD in theyounger subjects were still in the 3- to 6-fold range
and statistically significant; however, our precision de-
creased, probably because of correlated exposures. Correla-
tion between pesticides is an inherent problem when
assessing the effects of human exposure. However, since
adjustment for other pesticides did not remove the associa-
tion for maneb and paraquat, our data provide compelling
evidence that these 2 pesticides may in fact affect PD risk in
humans, as has been suggested by animal experiments.
Paraquat and maneb are applied by ground, aerial, and
backpack methods; however, paraquat has a much longer
field half-life of 1,000 days (33), as compared with only
12–36 days for maneb (34). Both chemicals bind strongly
to soil, though, and are not thought to be a threat to ground-
water (35, 36). Such strong binding could result in contam-
inated soil getting blown or tracked into homes by wind,
pets, and shoes, thereby increasing exposure for persons
who live closer to agricultural application sites (3, 37, 38).
In a previous validation study, our prediction model for a
serum measure of dichlorodiphenyldichloroethylene (DDE)
explained 47% of the biomarker’s variance (39). Addition-
ally, our GIS-derived measure of organochlorine exposure
identified persons with high serum DDE levels reasonably
well (specificity of 87%) (39).
Although our GIS model allowed us to calculate the num-
ber of pounds of each active ingredient applied per acre
within a 500-m buffer, these quantities are not comparable
across pesticides. That is, a pound of active ingredient does
not represent the same human neurotoxicity across pesti-
cides, and no information currently exists that would allow
us to standardize these measures. Thus, while we believe
that our model provided us with an accurate indicator of any
pesticide exposure from applications close to a residence,
our exposure measure cannot be considered quantitative
beyond a crude rank ordering of low/medium likelihood
of exposure and high likelihood of exposure. Since we hy-
pothesized that coexposure to 2 pesticides, maneb and para-
quat, would increase the risk of PD, we also lacked the
statistical power to perform extensive categorical analyses
(note that only 3 cases and 1 control were exposed solely to
maneb). We conducted additional analyses after dichoto-
mizing pounds per acre at their median and mean levels
and found that exposure to both pesticides at the highest
level was associated with PD, especially in persons aged
?60 years; however, wide confidence intervals surrounding
our point estimates rendered these results generally uninfor-
mative (results not shown).
In only 1 previous analysis, conducted within the Agri-
cultural Health Study cohort (40), did researchers assess the
effects of maneb and paraquat exposures. Statistical power
was limited by the small number (n ¼ 78) of incident cases
identified during follow-up and the very small number (n ¼
4–10)of cases exposedto maneb/mancozeb(OR ¼ 2.1) and
paraquat (OR ¼ 1.4). In a small Taiwanese study, the only
case-control study to date with sufficient statistical power to
examine exposure to the herbicide paraquat, Liou et al. (41)
reported a 4- to 6-fold increase in PD risk among long-term
applicators. In a case-control study from the Mayo Clinic
(Rochester, Minnesota), Brighina et al. (42) presented asso-
ciations between self-reported pesticide exposure and PD in
subjects younger than 60 years only (for all pesticides,
OR ¼ 1.80, 95% CI: 1.12, 2.87; for herbicides, OR ¼ 2.46,
95% CI: 1.34, 4.52).
Our exposure estimates did not depend on the subject’s
recall of pesticide exposure and are therefore unlikely to
have been biased by differential exposure misclassification.
Since all of our PD diagnoses were clinically confirmed, we
expect disease misclassification to have been minimal. Non-
differential exposure misclassification is a possibility in our
study and may have attenuated our effect estimates.
Our results may be biased if cases and controls selected
themselves into our study according to their potential for
pesticide exposure, but our subjects were not asked to self-
report environmental exposures and probably were unaware
of their true historical exposures. There is no reason to
suspect that cases and controls would have chosen to par-
ticipate on the basis of their historical residence near certain
agricultural plots. We saw no difference in estimated effects
when we restricted analyses to only those subjects with
more (?12 years) or less (<12 years) education. Similarly,
we saw no difference in our results when we restricted the
sample to persons whose addresses had been mapped with
high precision in the tricounty area during the period 1974–
1999 (363 cases, 336 controls).
Our analysis has confirmed 2 previous observations from
animal studies: 1) exposure to multiple chemicals may po-
tentiate the effect of each chemical (of interest, since hu-
mans are often exposed to more than 1 pesticide in the
environment) and 2) the timing of exposure is important.
To our knowledge, this is the first epidemiologic study to
provide strong evidence that 2 specific pesticides, suggested
by animal research as potentially acting synergistically to
become neurotoxic, strongly increase the risk of PD in
humans, especially given combined exposure and when
encountered earlier in life.
Author affiliations: Department of Environmental Health
Sciences, School of Public Health, University of California,
924Costello et al.
Am J Epidemiol 2009;169:919–926
Berkeley,Berkeley, California (Sadie Costello); Department
of Preventive Medicine, Keck School of Medicine, Univer-
sity of Southern California, Los Angeles, California (Myles
of Letters, Arts and Sciences, University of Southern
California, Los Angeles, California (Myles Cockburn,
Xinbo Zhang); Department of Neurology, School of Medicine,
University of California, Los Angeles, Los Angeles, Cali-
fornia (Jeff Bronstein); and Department of Epidemiology,
School of Public Health, University of California, Los
Angeles, Los Angeles, California (Beate Ritz).
This work was supported by the National Institute
U54ES12078, and 5P30 ES07048), the National Institute of
Neurological Disorders and Stroke (grant NS 038367), and
the Department of Defense Prostate Cancer Research Pro-
gram (grant 051037). In addition, initial pilot funding was
provided by the American Parkinson’s Disease Association.
The authors thank the participating neurologists and med-
ical centers in Fresno, Kern, and Tulare counties for their
Conflict of interest: none declared.
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