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Honey Bee (Apis mellifera) Exposure to Pesticide Residues in Nectar and

Pollen in Urban and Suburban Environments from Four Regions of the United
States

Article in Environmental Toxicology and Chemistry · January 2022

DOI: 10.1002/etc.5298

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Environmental Toxicology and Chemistry—Volume 00, Number 00—pp. 1–13, 2022
Received: 15 January 2021 | Revised: 17 February 2021 | Accepted: 19 January 2022 1

Environmental Chemistry

Honey Bee (Apis mellifera) Exposure to Pesticide Residues

in Nectar and Pollen in Urban and Suburban Environments
from Four Regions of the United States
Fabien J. Démares,a,b,1 Daniel Schmehl,c,d Jeffrey R. Bloomquist,a Ana R. Cabrera,c Zachary Y. Huang,e Pierre Lau,f,g
Juliana Rangel,f Joseph Sullivan,h Xianbing Xie,e,i and James D. Ellisd,*,1
a
Entomology and Nematology Department, Emerging Pathogens Institute, University of Florida, Gainesville, Florida, USA
b
Centre d'Écologie Fonctionnelle et Évolutive, Université de Montpellier, Centre National de la Recherche Scientifique, Ecole Pratique des Hautes Etudes,
Institut de Recherche pour le Développement, Montpellier, France
c
Bayer CropScience, Chesterfield, Missouri, USA
d
Honey Bee Research and Extension Laboratory, Entomology and Nematology Department, University of Florida, Gainesville, Florida, USA
e
Department of Entomology, Michigan State University, East Lansing, Michigan, USA
f
Department of Entomology, Texas A&M University, College Station, Texas, USA
g
US Department of Agriculture, Stoneville, Mississippi, USA
h
Ardea Consulting, Minford, Ohio, USA
i
Department of Laboratory Animal Science, Nanchang University, Nanchang, Jiangxi, China

Abstract: The risk of honey bee (Apis mellifera L.) exposure to pesticide residues while foraging for nectar and pollen is
commonly explored in the context of agroecosystems. However, pesticides are also used in urban and suburban areas for
vegetation management, vector control, and the management of ornamental plants in public and private landscapes. The
extent to which pesticides pose a health risk to honey bees in these settings remains unclear. We addressed this at a
landscape scale by conducting pesticide residue screening analyses on 768 nectar and 862 pollen samples collected monthly
over 2 years from honey bee colonies located in urban and suburban areas in eight medium to large cities in California,
Florida, Michigan, and Texas (USA). A risk assessment was performed using the US Environmental Protection Agency's
BeeREX model whenever an oral toxicity value was available for a compound. Chemical analyses detected 17 pesticides in
nectar and 60 in pollen samples during the survey. Approximately 73% of all samples contained no detectable pesticide
residues. Although the number of detections varied among the sampled regions, fewer pesticides were detected in nectar
than in pollen. Per BeeREX, four insecticides showed a potential acute risk to honey bees: imidacloprid, chlorpyrifos, and
esfenvalerate in nectar, and deltamethrin in nectar and pollen. In general, exposure of honey bees to pesticides via nectar
and pollen collection was low in urban and suburban areas across the United States, and no seasonal or spatial trends were
evident. Our data suggest that honey bees are exposed to fewer pesticides in developed areas than in agricultural ones.
Environ Toxicol Chem 2022;00:1–13. © 2021 SETAC

Keywords: Fungicide; Herbicide; Insecticide; Miticide; Urban landscape; Apis mellifera

INTRODUCTION
The western honey bee (Apis mellifera L.) is the most
commonly managed bee species in the world and is used to
provide pollination services in major cropping systems
This article contains online‐only Supporting Information.
1
Fabien J. Démares and James D. Ellis contributed equally to this work.
* Address correspondence to jdellis@ufl.edu
Published online 25 January 2022 in Wiley Online Library
(wileyonlinelibrary.com).
DOI: 10.1002/etc.5298
throughout the United States (Delaplane & Mayer, 2000) and
globally. Honey bees are also the backbone of an econom-
ically important industry composed of backyard, sideline, and
commercial beekeepers, some of whom keep their bees in
urban and suburban areas at least part of the year (Gallant
et al., 2014; Losey & Vaughan, 2006; Morse & Calderone,
2000). Managed honey bee colonies currently face different
stressors that affect their health. These include nutritional
deficiencies, pests, pathogens, poor queen quality, and
pesticides (Brodschneider & Crailsheim, 2010; Kulhanek
et al., 2017; Pirk et al., 2016). Among these factors, the role of

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2 Environmental Toxicology and Chemistry, 2022;00:1–13—F.J. Démares et al.
pesticides in the observed decline of honey bee health has
received considerable attention in the last decade (Chauzat
et al., 2009; Johnson et al., 2010; Kulhanek et al., 2017). As a
response, regulatory agencies worldwide have developed
detailed pesticide risk assessment frameworks for pollinators,
with A. mellifera serving as the predominant model/surrogate
organism for these assessments (European Food Safety
Authority [EFSA], 2013a; US Environmental Protection
Agency [USEPA], 2014).
The risk to bees that arises from pesticide exposure is a
function of both toxicity and exposure (Fischer & Moriarty,
2014). Toxicity is innate and is informed by laboratory and field
effect studies. Bee exposure to pesticides via pollen and nectar
collection depends on various factors including the physical
and chemical properties of the pesticide, bloom period of
treated plants, and application rates, methods, and timing
(USEPA, 2014, 2016a, 2016b; Fischer & Moriarty, 2014). These
factors may individually or concomitantly influence the sys-
temicity and environmental fate of a compound.
Most research efforts to date have focused on charac-
terizing honey bee exposure to pesticides in agricultural
settings (Calatayud‐Vernich et al., 2018; Mullin et al., 2010;
Ostiguy et al., 2019; Tosi et al., 2018), and rightly so, given
that agroecosystems are where most of the pesticide use
occurs for the control of food and feed crop pests (Brain &
Anderson, 2019; Popp et al., 2013). However, pesticides are
also used in nonagricultural settings such as homes, gar-
dens, and golf courses, as well as for vegetative manage-
ment and vector control (Brain & Anderson, 2019; Held &
Potter, 2012). Furthermore, a characteristic of urban and
suburban environments is the heterogeneity of floral re-
sources, including flowering trees, shrubs, potted plants, and
weeds (Bates et al., 2011; Čepelová & Münzbergová, 2012),
many of which can be treated with pesticides during bloom
(Losey & Vaughan, 2006).
Given the limited availability of pesticide use data in
developed landscapes, we conducted a study to determine
the exposure and risk to managed honey bees from pesti-
cides used in urban and suburban areas of the United
States. In particular, we were interested in the potential risk
to honey bees when colonies were exposed through nectar
and pollen collection. Consequently, we conducted a 2‐year
survey (July 2014 to June 2016) of nectar and pollen sam-
ples collected from colonies located in California (CA),
Florida (FL), Michigan (MI), and Texas (TX) to characterize
pesticide exposure at the developed landscape level. In
addition, when toxicity data were available for detected
compounds, we used the BeeREX model developed by the
USEPA's Office of Pesticide Programs (USEPA‐OPP) to
identify cases in which acute risks to bees may require
mitigation or further data generation (USEPA, 2021). The
present study constitutes a starting point in identifying po-
tential spatial and/or temporal trends for honey bee ex-
posure to pesticides in urban and suburban environments. It
also allows us to identify, at a landscape scale, specific ex-
posure scenarios in developed areas that pose risks to bees
and might warrant further investigation.
© 2022 SETAC
MATERIALS AND METHODS
Hive locations
Colonies managed by volunteer beekeepers were identi-
fied in urban and suburban areas in CA (San Francisco/Bay
Area and Sacramento), FL (Tampa Bay Area and Orlando),
MI (Detroit and Lansing), and TX (Austin and Bryan).
The criteria for accepting a colony into the study included:
(1) a minimum area of primary land development sur-
rounding the apiary, (2) a distance of at least 3.2 km be-
tween focal study colonies to maintain independence among
samples, and (3) having at least two colonies in the same
location to have a replacement in case the monitoring
colony died or became too weak for monthly collections. If
beekeepers ended their participation in the study, their
colonies were replaced by colonies from other volunteers
following the same criteria. In all, 20, 19, 15, and 18 apiary
locations were used for sampling in CA, FL, MI, and TX,
respectively. More information on site colony selection can
be found in Lau et al. (2019).
Land‐use analysis
A land‐use analysis was conducted around each colony
using ArcGIS Ver 9.2 and Ver 10.4. There were two primary
reasons for performing this analysis. First, we wanted to
ensure that colonies were not close to land used for agri-
cultural purposes. Second, we wanted to determine the
amount of developed land near the colonies to confirm that
they were located in urban/suburban areas. The 2011 anal-
ysis included the area around each colony at radii of 0.8,
1.6, and 4 km. The National Land Cover Database (NLCD)
was used to classify the land cover types around each
colony. The NLCD is a Landsat‐based, 30‐m resolution land
cover database for the entire United States. The NLCD
products are created by the Multi‐Resolution Land
Characteristics (MRLC) Consortium, a partnership of Federal
agencies led by the US Geological Survey. All NLCD
data products are available for download at no charge to
the public from the MRLC website (2021). This dataset was
supplemented with recent aerial images for the areas
around each colony by overlaying the aerial images and
manually reclassifying changes in land cover development
to encompass the recent development in suburban/
urban areas. The categories of land use included in the
analysis were: developed–high intensity, developed–medium
intensity, developed–low intensity, developed–open space,
recently disturbed or modified, cultivated cropland, forest
and woodland, shrubland and grassland, wetland systems,
aquatic, and other. The analysis focused on two categories
of land cover classes: developed areas and agricultural
areas. For purposes of characterizing the area surrounding
each study colony as urban/suburban, the four developed
categories were pooled, and the remaining categories
were pooled as nondeveloped, as was done in Lau
et al. (2019).
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Honey bee exposure to pesticides in urban environments—Environmental Toxicology and Chemistry, 2022;00:1–13
Sample collections
Colonies were sampled monthly during the standard bee-
keeping season for each state (22 months over 2 years in CA;
24 months over 2 years in FL; 13 months over 2 years in MI; and
12 months over 2 years in TX; Supporting Information,
Table S1). Pollen traps (Brushy Mountain Bee Farm's item
ND#464 for 10‐frame hives, or Brushy Mountain Bee Farm's
item #509 for 8‐frame hives) were activated at the entrance of
each hive approximately 2–3 days prior to collecting pollen
samples. In addition, a frame containing an empty drawn comb
was placed inside the honey super of each hive 1–7 days prior
to sample collection to provide an area from which fresh nectar
could be collected. Pollen was collected from the traps and
placed into a 50‐ml plastic centrifuge tube. Nectar was col-
lected from the designated comb using a disposable plastic
pipette and transferred into a 50‐ml plastic centrifuge tube.
The pipettes and tubes originated from newly opened pack-
ages and were not rinsed or cleaned prior to use. The bees did
not always store nectar in the frame provided to them. In these
few instances, uncapped nectar was collected from neigh-
boring combs in the same hive. Honey bees may move honey/
nectar from the brood area into empty combs placed in the
honey super. Thus, although our intention was to collect fresh
nectar, we can only confidently say that it was uncapped nectar
and/or honey.
The tubes were placed on ice in a cooler and transported
from the field to the laboratory where the pollen and nectar
samples were weighed. Samples were then stored at −20 °C
until they were sent off for chemical analysis. Pollen samples
were separated into two parts: a 3‐g sample was used for
pesticide residue screening, and a 1‐g sample was sent to
colleagues at Texas A&M University (College Station, TX, USA)
for purposes of plant taxonomic identification (Lau et al., 2019).
Priority was given to the pesticide residue analysis when not
enough pollen was collected for both pesticide and plant
source analyses. In total, 275, 273, 190, and 124 pollen sam-
ples were collected from colonies in CA, FL, MI, and TX, re-
spectively. In addition, 224, 264, 193, and 87 nectar samples
were collected in CA, FL, MI, and TX, respectively (Supporting
Information, Table S1).
Chemical analysis
Nectar and pollen samples were submitted periodically to
the US Department of Agriculture–Agricultural Marketing
Service–National Science Laboratories (USDA–AMS–NSL)
facility in Gastonia, North Carolina for the Apiculture Pesticide
Screen analysis that quantifies residues for 173 compounds,
including pesticide active ingredients and metabolites. These
represent pesticides of interest for honey bee toxicology and
active ingredients to which honey bees are likely to be ex-
posed. The methodology for the multiresidue analysis, in-
cluding the use of high‐purity standards, was previously
described in detail in Mullin et al. (2010). It is based on a
modified quick, easy, cheap, effective, rugged, and safe
(QuEChERS) method (Lehotay et al., 2005) analyzed via liquid
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3
chromatography–tandem mass spectrometry, allowing for the
detection of pyrethroids, whereas other methods may not.
Pesticides were classified into four main categories: in-
secticides (89), fungicides (39), herbicides (27), and miticides/
acaricides (13). The list also included one inert ingredient,
three synergists, and one repellent. Levels of detection (LODs)
ranged from 1 to 100 ppb depending on the pesticide
(Supporting Information, Tables S2 and S3). It is important to
note that the LOD for some pesticides could be higher than the
level of concern (LOC).
Pollinator risk assessment
The BeeREX model was developed by the USEPA‐OPP and
is used as the first step in the risk assessment process for
pesticide exposure. It calculates a risk quotient for any pesti-
cide based on dietary exposure and contact exposure for
honey bees at different ages and classes (USEPA, 2014). The
BeeREX model considers the primary routes of dietary and
contact exposure to bees, with the application rate and
method of application as the starting point for exposure. When
exposure is refined via residue analysis of bee‐relevant matrices
as in our study, the total dose is calculated by multiplying the
estimated environmental concentration (i.e., the detected res-
idues in pollen and/or nectar) by the estimated dietary con-
sumption for a particular age of bee. For example, BeeREX
estimates that forager bees consume 292 mg of nectar/day and
0.041 mg of pollen/day. In our study, the maximum detected
residue was converted to a total dose based on dietary con-
sumption estimates, and then was divided by the acute oral
median lethal dose (LD50) value to derive a risk quotient (RQ).
Values of RQ above 0.4 for an acute route of exposure indicate
a potential risk of the pesticide at the laboratory screening level
for individual bees. This risk characterization may require the
need for additional data, especially frequency of exposure, at
the colony level (i.e., increased realism) to characterize the risk
of a particular pesticide at the colony level or develop miti-
gation options such as changes to the label for the use of a
product. The calculated RQs indicate acute risks by con-
sumption of residues in food (i.e., short exposures). The risks of
chronic exposure to such residues are higher for most com-
pounds. Although not possible currently, our residue data can
be used to calculate chronic RQs when chronic toxicity data
become publicly available.
RESULTS
Landscape analysis
The proportion of developed land near the colonies varied
somewhat across the different regions. Some study locations,
particularly those in coastal FL and CA, were near marine or
estuarine habitats. To estimate the proportion of managed
landscapes near each colony, eliminating the aquatic habitat
provided a better idea of the proportion of foraging habitat
that was developed and might be treated with pesticides. More
than 80% of the landscape within 1.6 km of the colonies and
© 2022 SETAC
4 Environmental Toxicology and Chemistry, 2022;00:1–13—F.J. Démares et al.
more than 75% of the landscape within 4 km of the colonies
was classified as developed when the aquatic habitats were
excluded. Within 1.6 km of the colonies, developed lands
ranged from 52% to 100%, 57% to 100%, 58% to 100%, and
13% to 100% in CA, FL, MI, and TX, respectively. Within 4 km of
the colonies, developed lands ranged from 40% to 99%, 30%
to 99%, 39% to 100%, and 42% to 97% in CA, FL, MI, and TX,
respectively. Colonies in CA and FL had essentially no cropland
(one colony in both regions had 0.2% cropland) within 4 km. In
MI, cropland (up to 10.8%) existed within 4 km of seven colo-
nies. Three colonies in TX had cropland within 4 km, with one
site showing 2.0% cropland.
Pesticide residue analysis
A total of 1630 samples were collected and analyzed across
the four states over 24 months (Supporting Information,
Table S1), making this the largest pesticide residue screening
for honey bee colonies in urban and suburban landscapes to
date. Of 768 nectar samples collected throughout the study,
654 (85.2%) had no pesticide residues above their respective
LOD, whereas 114 (14.8%) had one or more pesticide residues
detected (Figure 1A). Comparatively, 534 of 862 (61.9%) pollen
samples had no pesticide residues above their respective
LODs, whereas 328 (38.1%) samples contained one or more
pesticides (Figure 1A). Of the 114 residue‐positive nectar
samples, 89 samples (78.1%) had only one compound de-
tected, 16 (14%) and 9 (7.9%) samples had two and three
compounds detected per samples respectively, and none had
more than three residues per sample (Figure 1B).
Of the 328 residue‐positive pollen samples, 189 samples
(57.6%) had one compound detected. In addition, two and
three residues per sample were detected in 88 (26.8%) and
FIGURE 1: Survey of pesticides detected in nectar and pollen samples
collected from US honey bee hives. (A) Total number of samples
containing one or more pesticides (nectar: green; pollen: blue), or
those with no detectable pesticide residues (gray). The number of
samples collected in each category is indicated inside the bars. (B) The
number of nectar (top graph) and pollen (bottom graph) samples with
positive detection of pesticide residues. The number of residue‐
positive samples containing 1, 2, 3, or more individual residues is in-
dicated above each bar.
© 2022 SETAC
38 (11.6%) of the samples analyzed, respectively. Only 13 (4%)
pollen samples had four or more compounds detected. Of the
173 pesticides screened, 63 (36.4% of the total) were detected
at least once in bee‐collected nectar and/or pollen across the
four states during the 2‐year survey (Supporting Information,
Tables S2 and S3). Of these, three pesticides were detected
only in nectar samples, 46 were detected only in pollen sam-
ples, and 14 were found in both nectar and pollen samples
(Supporting Information, Table S4).
The 63 pesticides detected through the analytical screen
were divided into four classes: fungicides (18 detected), her-
bicides (11), insecticides (29), and miticides (9). Four com-
pounds belonged to two classes: thymol, which is both a
miticide and a fungicide; and bifenthrin, coumaphos, and flu-
valinate, which are used as miticides and insecticides. When we
considered all four states together, insecticides were the most
commonly detected class of compound in both nectar samples
(121 out of 148 residue detections) and pollen samples (275
out of 537 residue detections). For nectar samples, miticides
were the second most detected class (41 out of 148 residue
detections), whereas for pollen samples, herbicides were
the second most detected class (160 out of 537 residue
detections). Interestingly, the distribution and ranking of
pesticide classes differed among individual states (Supporting
Information, Table S4).
For nectar samples in CA urban areas, insecticides (38 de-
tections out of 39 total) were more represented than were
herbicides (1), and no miticide or fungicide residues were de-
tected. In FL urban areas, insecticides (10 detections out
of 24 total), miticides (17), and fungicides (4) were detected,
whereas herbicides were not detected. Insecticides were the
most common pesticides detected in MI nectar samples
(73 detections out of 81 total), followed by miticides (24), and
then fungicides (4). In TX urban areas, the opposite situation
was observed, with only fluridone being detected in nectar
samples, making herbicides (four detections in total) the only
class found in TX nectar (Supporting Information, Table S4).
For pollen samples, each pesticide class was detected in
different numbers for every state. In CA, insecticides were the
most frequently detected pesticide residue class (99 detections
out of 165), followed by fungicides (38), herbicides (24), and
miticides (15). Pollen samples from FL and MI urban areas
showed the same class rankings, with insecticides being de-
tected most frequently (67 detections out of 148 in FL, and 100
detections out 179 in MI), followed by herbicides (FL: 51;
MI: 58), miticides (FL: 51; MI: 22), and fungicides (FL: 20; MI:
20). The observations were different in TX urban areas, where
the major presence of fluridone made herbicides the most
frequently detected pesticide class (27 detections out of 45).
This was followed by insecticides (9), fungicides (9), and miti-
cides (9; Supporting Information, Table S4).
Pesticide detection across states
The number of nectar and pollen samples collected varied
by state due to different climatic conditions and local floral
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Honey bee exposure to pesticides in urban environments—Environmental Toxicology and Chemistry, 2022;00:1–13 5

resources. For example, nectar samples in FL were collected
year‐round, whereas nectar collection in CA was interrupted in
December or January. The winter season was longer in MI. In
addition, we began collecting samples in TX later in the study,
resulting in only 12 months of collection during the 2‐year
survey (Supporting Information, Table S1). Thus, more nectar
samples were collected in FL and CA (264 and 224 samples,
respectively) than in MI and TX (193 and 87, respectively).
Nevertheless, a higher percentage of residue‐positive nectar
samples was found in MI (31.1%) compared with those of the
other three states (CA: 12.1%; FL: 8.7%; and TX: 4.6%;
Figure 2A). In addition, most of the residue‐positive nectar
samples in each state contained only one pesticide (CA: 74.1%;
FL: 95.7%; MI: 71.6%; and TX: 100%; Figure 2B).
Similarly, more pollen samples were collected in FL and CA
(273 and 275 samples, respectively) than in MI and TX (190 and
124, respectively). A higher percentage of residue‐positive
pollen samples was found in MI (52.6%) compared with those
of the other three states (CA: 34.2%; FL: 35.9%; and TX: 29%;
Figure 3A). In total, the number of residue‐positive pollen
samples (328 vs. 114), the total number of positive detections
(537 vs. 148), and the different compounds detected (60 vs. 17)
were greater for pollen samples than for nectar samples, re-
spectively (Figure 1 and Supporting Information, Table S4).
Most of the residue‐positive pollen samples only contained one
pesticide (CA: 51.1%; FL: 63.3%; MI: 48%; and TX: 86.1%), and
when positive, they were found at a lower percentage com-
pared with residue‐positive nectar samples. This was due to a
higher number of residue‐positive samples containing two or
more pesticides in pollen samples compared with nectar
samples (Figure 3B).
Pesticide screening and identification
Imidacloprid was the most consistently detected compound
in nectar among the 17 different pesticides detected (Table 1
and Supporting Information, Tables S3 and S4). This neon-
icotinoid appeared in nectar in three states (CA, FL, and MI),
with a maximum detection of 25 imidacloprid‐positive nectar
samples (11.2%) out of 224 total samples analyzed in CA
(64.1% of 39 total pesticide detections in CA). Furthermore,
esfenvalerate was the most detected residue in MI, with 50
positive detections (25.9%) in nectar samples out of 193 total
samples analyzed (61.7% of 81 total pesticide detections in MI).
In addition, we obtained positive detections (but less fre-
quently) in MI for the organophosphate chlorpyrifos (1 positive
sample, or 0.5%) and the pyrethroids deltamethrin (2 positive
samples, or 1%) and fluvalinate (11 positive samples, or 5.7%)
out of 193 nectar samples analyzed (respectively, 1.2%, 2.5%,
and 13.6% of 81 total pesticide detections).
Single pesticide detections (189 pollen samples with one
pesticide residue) occurred more often in pollen (862 total
pollen samples) than did multiple pesticide detections
(139 pollen samples with multiple pesticides). California had
the highest unique‐detect count (35 different pesticides de-
tected) compared with the rest of the states: FL (21), MI (29),
and TX (14). Imidacloprid, the most common pesticide found in
CA pollen, was detected in 20.7% of the analyzed samples
(57 detections out of 275 total samples, 34.5% out of 165 total
pesticide detections). Imidacloprid was also detected in 5.5%
of pollen samples collected in FL (15 detections out of 273 total
samples, 10.1% of 148 total pesticide detections), 4.7% of
those collected in MI (9 detections out of 190 total samples, 5%
of 179 total pesticide detections), and 0.8% of those collected
in TX (1 detection out of 124 total samples, 2.2% of 45 total
pesticide detections). Also, the herbicide fluridone was the
most commonly detected compound in TX, with 21 detections
(16.9%) out of 124 samples analyzed (46.7% of 45 total pesti-
cide detections), and it was not detected in samples from other
states. Esfenvalerate was the most commonly detected pesti-
cide in pollen samples from MI, with 50 positive detections
(26.3%) out of 190 samples analyzed (27.9% of 179 positive

FIGURE 2: Survey of pesticides detected in nectar samples collected from US honey bee hives. (A) The number of samples containing one or more
pesticides in nectar (green), or no detectable pesticide residues (gray) in California (CA), Florida (FL), Michigan (MI), and Texas (TX). The number of
nectar samples collected per category is indicated within the bars for each state. (B) The state‐specific number of samples with positive residue
detection within each frequency category. Graphs from left to right, top to bottom, represent data from CA, FL, MI, and TX. For each state, the
number of residue‐positive nectar samples containing 1, 2, 3, or more individual residues is indicated above each bar.

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6 Environmental Toxicology and Chemistry, 2022;00:1–13—F.J. Démares et al.

FIGURE 3: Survey of pesticides detected in pollen samples collected from US honey bee hives. (A) The number of samples containing one or more
pesticides in pollen (blue), or no detectable pesticide residues (gray) in California (CA), Florida (FL), Michigan (MI), and Texas (TX). The number of
pollen samples for each category is indicated within the bars for each state. (B) The state‐specific number of samples with positive residue detection
within each frequency category. Graphs from left to right, top to bottom, represent data from CA, FL, MI, and TX. For each state, the number of
residue‐positive pollen samples containing 1, 2, 3, or more individual residues is indicated above each bar.

detections). Other residues detected in pollen samples from all
four states included bifenthrin, carbendazim, chlorpyrifos,
pendimethalin, and thymol.
Pollinator risk assessment
We report the mean concentration of residues in samples
with positive pesticide detections in the Supporting In-
formation, Table S2. If the LOD was not reached in a sample,
the residue concentration value was assumed to be half of the
highest LOD when the mean was calculated across all analyzed
samples. The maximum residue concentrations detected were
used as exposure estimates to calculate an acute RQ via the
USEPA's risk assessment tool BeeREX. For all pesticide resi-
dues for which an oral LD50 was publicly available, an RQ value
was calculated and is reported in the Supporting Information,
Table S2. Twenty‐two detections (of 685 detections in 1630
samples) representing four compounds reached a calculated
acute RQ value above an LOC of 0.4, indicating potential acute
risk to honey bees (Table 1). Therefore, per the USEPA risk
assessment process, these compounds require further evalua-
tion of risks in higher tier tests. All were insecticides detected at
different levels, mostly within nectar, with one being present in
pollen. Imidacloprid (9 samples exceeding the LOC) was de-
tected at different mean concentrations in CA (0.85 ppb), FL
(0.54 ppb), and MI (0.62 ppb) samples. These mean concen-
trations were lower than the LOD for imidacloprid (1 ppb) be-
cause more than 90% of the collected samples did not contain
any detectable imidacloprid residues. When the mean con-
centrations in samples with positive detects were calculated (as
was done in Mullin et al., 2010), imidacloprid levels exceeded
the LOD in CA (3.6 ppb, 25 detects out of 224 samples), FL
(5.4 ppb, two detects out of 264 samples), and MI (8.4 ppb,
three detects out of 193 samples). The RQ values based on the
maximum detection of imidacloprid in nectar were 0.85 in CA,
0.42 in FL, and 0.96 in MI. The organophosphate chlorpyrifos
(1 detect out of 193 samples, 0.5% detection rate) and the
pyrethroid esfenvalerate (50 detects out of 193 samples, 25.9%
detection rate, 7 samples exceeding the LOC) were detected in
nectar samples in MI at levels above their LOC of 342 and
1095 ppb, respectively. The calculated RQ values based on the
maximum detection of chlorpyrifos and esfenvalerate in nectar
were 0.62 and 6.5, respectively. Another pyrethroid insecticide,
deltamethrin, was detected in MI at concentrations above the
LOC in both nectar (2 samples) and pollen (3 samples) in 1.3%
of the total samples (five detects in 383 pollen/nectar samples).
The calculated RQ values varied considerably based on the
dietary route of exposure (bees consume more nectar than
pollen), with an RQ value of 69.88 for deltamethrin in nectar
and 1.35 in pollen despite the overall similar concentrations
that we detected in the nectar and pollen samples.
Seasonality analysis
We present residue data composited by month and season
for each state in the Supporting Information, Table S5. Overall,
residue‐positive nectar samples were more commonly found
from April to October in most states. Residue‐positive nectar
samples were more common during spring in CA, summer in
FL, spring in MI, and winter/spring in TX. No apparent overall
seasonal pattern was evident for residue‐positive pollen sam-
ples. Nevertheless, residue‐positive pollen samples were more
common during summer in CA, summer/autumn in FL, spring in
MI, and spring in TX (Supporting Information, Table S5).
Individual colony exposure over time
Our data permitted us to describe in detail the repeated
exposure of individual honey bee colonies to pesticides in
nectar and pollen monthly at levels above and below the LOC.
Those data are reported in the Supporting Information,

© 2022 SETAC wileyonlinelibrary.com/ETC

Honey bee exposure to pesticides in urban environments—Environmental Toxicology and Chemistry, 2022;00:1–13 7

Table S6, for exposure through nectar and in the Supporting

parts per billion [ppb]), total number (no. of samples with residues) and percentage (% samples with residues) of samples with residues, mean (overall and of detected samples), maximum detection (max.), level of concern

Oral LD50 values for acute exposure of honey bees were obtained from public databases from the US Environmental Protection Agency (USEPA), the North Central Integrated Pest Management Center, and France's
Mean overall: If a sample had a residue level lower than the limit of detection (LOD), half of the highest LOD was assumed for mean calculation purposes. Correspondingly, this is the mean across all samples, including
The data include the number of samples collected and number of colonies sampled in each state (California, CA; Florida, FL; Michigan, MI; Texas, TX), sample type (nectar or pollen), compound, limit of detection (LOD; in

(LOC), no. of samples that exceeded the LOC (no. of colonies with samples exceeding the LOC), reported oral LD50 (lethal dose that kills 50% of the population, expressed in µg active ingredient [ai]/bee), and calculated

BeeREX: Terrestrial model for pesticide risk assessment developed by the USEPA, Canada's Pest Management Regulatory Agency, and the California Department of Pesticide Regulation (USEPA, 2021). RQ values ≥ 0.4
RQ‐BeeREXe
Information, Table S7, for exposure through pollen. In all, 14 of

0.85
0.42
0.96
0.62

69.88
1.35
6.5
74 colonies (18.9%) in the study had at least one residue that
exceeded the LOC, whereas 5 colonies (6.8%) had two residues
that exceeded the LOC. The percentages were particularly
Oral LD50d

high in MI, where 9 of 15 colonies had at least one residue that

0.0038
0.0038
0.0038

0.0015
0.0015
(µg ai/
bee)

0.25
exceeded the LOC (60% of all colonies) and 5 colonies (33%)
0.8 had two compounds that exceeded the LOC. Only three col-
onies throughout the study (one with imidacloprid in nectar in
No. of samples >

samples > LOC)

both CA and FL, one with esfenvalerate in nectar in MI) had

colonies with
LOC (no. of

residues that exceeded the LOC in nectar during back to back

(4)
(1)
(2)
(1)
(6)
(2)
(3)

months. No similar pattern was seen for residues in pollen.

5
2
2
1
7
2
3

In all, 5 colonies in CA (25% of all colonies), 3 in FL (15.8%),

and 15 in MI (100%) had residues detected in nectar over
multiple months. No colony had residues detected in nectar

indicate there is potential risk at the Tier I level (individual bees), and higher tier assessments are necessary to characterize risk to honey bees at the colony level.
over multiple months in TX. In contrast, 13 colonies in CA
c

(ppb)

1095
LOC

342
5.2
5.2
5.2

62
2

(65%), 17 in FL (89.5%), all in MI (100%), and 12 in TX (60%) had

residues detected in pollen over multiple months.
17800
(ppb)
Max.

12.5
532

359
211
5.5
11

DISCUSSION
detectedb
TABLE 1: Pesticides detected in nectar and pollen collected from US honey bee hives with risk quotient values ≥0.4

Mean

(ppb)

1026
532

259
194
3.6
5.4
8.4

The primary objective of our study was to focus on honey

bee colony exposure to pesticides in nectar and pollen in
managed and/or developed landscapes within urban/suburban
areas. To do this, it was important to confirm some basic cri-
risk quotient (RQ) using BeeREX (derived from the maximum detected residue). All residue levels are expressed in ppb.
overalla

teria. First, we needed to evaluate the level of land develop-

Mean

(ppb)

0.85
0.54
0.62

27.4
27.7
265

The LOC equates to the concentration in nectar or pollen that would result in an RQ = 0.4 (40% of the oral LD50).
3.2

ment that existed around each focal colony to confirm that the
criterion of land development for urban/suburban sites was
met. Colonies in each region were located in areas dominated
with residues
% Samples

by developed landscapes, averaging more than 80% devel-

11.2

25.9
0.8
1.6
0.5

1.6
1

opment within 1.6 km and more than 75% development within

4 km of each colony. Thus, our first assumption was met.
Second, it was important to eliminate or reduce croplands from
samples with

the studied landscapes so that we could definitively identify

residues

AGRITOX. If more than one LD50 value was available, the lowest one was selected.
No. of

n = 25

n = 50
n=2
n=3
n=1

n=2
n=3

exposure of honey bees to pesticides used in urban/suburban

landscapes rather than in agroecosystems. Colonies in CA and
FL had essentially no cropland nearby, and only a few colonies
in MI or TX had any cropland nearby, with no one colony being
(ppb)
LOD

Mean detected: This is the mean of residue‐positive samples only.

50
50

exposed to more than 11% of cropland. Given the foraging

1
1
1
1
2

radius of a honey bee colony of 6 km or further (Visscher &

Esfenvalerate
Deltamethrin
Deltamethrin
Imidacloprid
Imidacloprid
Imidacloprid

Seeley, 1982), it remains possible that some bees in the study

Chlorpyrifos
Compound

those where residues above the LOD were not detected.

occasionally visited croplands. However, we feel that in our

study the impact of agricultural pesticide application was
minimized, and likely eliminated, for a few key reasons. First,
the pesticides we detected are labeled for use in urban/
Sample

Nectar

Pollen

suburban settings within the United States. Second, an analysis

type

of the pollen collected by our focal colonies showed that bees

were foraging on plants that one would find in urban/suburban
areas, rather than on plants found in agricultural areas (Lau
State colonies sampled)
collected (no. of
No. of samples

et al., 2019). Third, there were no seasonal patterns to pesti-

(20)
(19)
(15)
(15)
(15)
(15)
(15)

cide residues in pollen, as one might expect if bees were for-

224
264
193
193
193
193
190

aging on agricultural land when plants are blooming, primarily

in the spring or early summer. The exposures were sporadic
and likely linked to local conditions that were not clear from
other types of information we obtained regarding the study
CA

MI
MI
MI
MI
MI
FL

sites (e.g., land‐use analysis). This, coupled with the geographic

b

e
a

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8 Environmental Toxicology and Chemistry, 2022;00:1–13—F.J. Démares et al.
information system analysis, supported our premise that most
of the pesticides detected in our nectar and pollen samples
had been applied in urban/suburban landscapes.
Next, we used BeeREX to calculate acute RQs, rather than
hazard quotients (HQs), for the residues found in nectar and
pollen in our study. Historically, HQs have been used to set a
benchmark to compare the exposure effects that pesticides can
have on different organisms, including honey bees (Frazier
et al., 2015; Mullin et al., 2010; Stoner & Eitzer, 2013). Hazard
quotients have also been used to calculate and identify risk to
an organism by comparing a concentration of a chemical to its
acute toxicity value (Ostiguy et al., 2019; Stoner & Eitzer, 2013).
However, this approach fails to convert the concentration to an
actual dose. In the United States, the BeeREX model is the
screening‐level pollinator risk assessment tool created by the
USEPA to translate the concentration of a pesticide to a dose
using an individual's nectar and pollen consumption estimates.
In our study, the resulting dose was divided by the acute oral
LD50 value to calculate an RQ, which, when compared to the
LOC, indicates the potential risk of a pesticide at the laboratory
screening level for individual bees. In addition, it has been
common practice when conducting pesticide surveys to ex-
clude nondetects when calculating the mean, median, and
percentile values for pesticide concentrations found in a
sample (see Mullin et al., 2010). Our analyses considered the
entire data set of samples when reporting the mean, median,
and 90th and 95th percentiles (Supporting Information,
Table S2) to represent the distribution of pesticide residues in
bee matrices within urban and suburban landscapes more
accurately.
Generating an acute HQ or RQ without considering ex-
posure frequency, route, or time‐cumulative effects does not
capture chronic and sublethal effects at the colony level, a
concern that has been expressed previously (Aupinel et al.,
2007; Thompson & Thorbahn, 2009). Indeed, there has been
growing support for chronic, rather than acute, risk assess-
ments of pollinator exposure to pesticides (EFSA, 2014; Halm
et al., 2006; Hesketh et al., 2016; Mommaerts et al., 2010;
Simon‐Delso et al., 2018; Spurgeon et al., 2016). We agree that
such assessments are warranted and provide additional data
to support the assessments done from an acute toxicity per-
spective. Nevertheless, we suggest that chronic risk assess-
ments for this type of study cannot be met easily for multiple
reasons. First, standardized methods to generate chronic
toxicity endpoints have only recently been developed
(Organisation for Economic Co‐operation and Development,
2016, 2017). Thus, the data are only just becoming available.
Quignot et al. (2015) found that few of the available studies
(~20%) report chronic effects, whereas the majority report acute
topical (61%) or oral (19%) toxicity data (Carnesecchi et al.,
2019). Second, using chronic HQs is inferior to using BeeREX‐
derived acute RQs because chronic HQs overestimate the
potential impact of pesticides to bees (Wen et al., 2021). The
acute RQs we calculated are based on the maximum measured
concentrations, thus providing a conservative approach to es-
timate risk across all colonies and locations. Ultimately, acute
RQs at the Tier 1 level trigger higher tier testing (semifield
© 2022 SETAC
and field conditions), whereby chronic impacts to bees are
screened. Third, the USEPA continues to use acute RQs values
for their assessments. Correspondingly, other authors continue
to publish research using acute RQ methods (see Traynor et al.,
2021; Tosi et al., 2018). Finally, and more related to our present
study, we noticed a random temporal variation of pesticide
exposure at each site. These exposures fluctuated significantly
on a month to month basis. Consequently, we argue that using
an acute oral assessment would be more accurate because of
the variability and overall low quantity of pesticides that we
detected in suburban/urban environments. Our raw data
(Supporting Information, Tables S6 and S7) are available for
future chronic RQ calculations as more chronic toxicity data
become publicly available.
Our data suggest that acute risk to honey bees from in-
dividual pesticide residues in nectar and pollen is generally low
in urban and suburban areas of the United States, at least in the
locations we tested. Although pesticide residues are generally
higher in pollen, the relative acute risk to bees is higher from
the nectar route of exposure because of a higher estimate of
nectar consumption compared with that of pollen. The BeeREX
model estimates that a forager bee consumes 292 mg of
nectar/bee/day on average, whereas the greatest pollen con-
sumption is estimated to be 9.6 mg/bee/day for nurse bees.
Those estimates were important factors for the risk character-
ization that we calculated. Furthermore, the BeeREX model
presents the highest RQ, because it uses either the RQ for
foraging bees or for nurse bees, whichever is higher. It is
important to note that the USDA NSL Gastonia Laboratory
Apiculture Pesticide analysis screened for 173 compounds (in-
cluding some metabolites) of nearly 1300 registered active in-
gredients at the time of the present publication. Nevertheless,
only a few of the compounds screened by the USDA laboratory
may pose a risk to honey bees at the levels found in hives when
considering exposure to one compound at a time. Of the 173
pesticide compounds included in the nectar and pollen anal-
ysis, 63 (36.4% of the total) were detected across our sampling
sites. All the detected pesticides were observed at rates lower
than 6% of all samples (Table 1 and Supporting Information,
Table S2), except for imidacloprid in CA, which was detected in
11.2% of all the nectar samples, and esfenvalerate in MI, which
was detected in 25.9% of all the nectar samples. From these
results, we compared existing toxicological data reported on
bees and calculated the acute risk associated with each com-
pound individually using the USEPA's BeeREX model. Only 4 of
the 63 pesticides detected were found to pose an acute risk to
individual honey bees by exceeding the LOC according to our
BeeREX screening‐level assessment. The four compounds were
chlorpyrifos, deltamethrin, esfenvalerate, and imidacloprid,
which represented 6.3% of the total number of pesticides de-
tected (4 out of 63) and 2.3% of the 173 pesticides included in
the USDA analysis.
The effects of imidacloprid on honey bee behavior and
metabolism have been extensively reported since the late
2000s (for review, see Godfray et al., 2014). Imidacloprid is a
member of the neonicotinoid class of insecticides, a class that
acts on the insect's central nervous system as an agonist of
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Honey bee exposure to pesticides in urban environments—Environmental Toxicology and Chemistry, 2022;00:1–13
nicotinic acetylcholine receptors (Matsuda et al., 2020). Ne-
onicotinoids affect many physiological processes in honey
bees, including flying ability, thermoregulation, sugar sensi-
tivity, learning, and memory (Démares et al., 2018; Henry et al.,
2012; Tosi et al., 2016; Williamson et al., 2013). Imidacloprid
has been found at different concentrations from agricultural
fields to urban and suburban areas, ranging from 0.1 to 10 ppb
in nectar, and from 6 to 51 ppb in pollen (Feltham et al., 2014;
Henry et al., 2015; Mullin et al., 2010; Ostiguy et al., 2019;
Stoner & Eitzer, 2012; also see Cresswell, 2011 for a meta‐
analysis on imidacloprid field‐realistic doses in nectar and
pollen). These values are consistent with our reported ob-
servations in urban and suburban areas (Table 1). Interestingly,
Dai et al. (2019) reported a calculated RQ value of 0.01 for
larvae reared in vitro and exposed to imidacloprid through
food. If we compare that RQ value with our calculated RQ
values for adult bees (from 0.42 to 0.96, Table 1), this suggests
that larvae could be at lower risk than adult bees when exposed
to imidacloprid.
Deltamethrin and esfenvalerate are type II pyrethroids and
act on voltage‐gated sodium channels, thus disturbing sodium
conductance and altering action potential generation
(Soderlund, 2010, 2012; Zlotkin, 1999). Pyrethroids impact
learning ability and foraging/homing flights in honey bees
(Decourtye et al., 2004; van dame et al., 1995). Both esfen-
valerate and deltamethrin were previously detected in pollen
(3.3–16.9 and 4.5–66 ppb, respectively), and deltamethrin was
detected in honey/nectar at 4.6 ppb in crop fields (Mullin et al.,
2010; Paradis et al., 2014; Pettis et al., 2013; and see Sanchez‐
Bayo & Goka, 2014 for a comprehensive meta‐analysis). In our
study, the mean detection value of deltamethrin in MI was
27.7 ppb for pollen and 27.4 ppb for nectar (Table 1). Although
deltamethrin was only detected in 2 of the 193 MI nectar
samples, the RQ was the highest reported (69.88) because the
associated LD50 value in honey bees is low (1.5 ng/bee), and
the BeeREX model does not consider the frequency of de-
tection. The reported mean values of esfenvalerate in nectar
and pollen detected in MI urban/suburban areas were 265 and
343 ppb, respectively (Supporting Information, Table S2),
which was much higher than the values reported in previous
field studies (Mullin et al., 2010; Ostiguy et al., 2019). Inter-
estingly, only the nectar route of exposure resulted in an RQ
higher than 0.4 (RQ = 6.5; Table 1), which could be explained
by the fact that nectar consumption values in the BeeREX
model result in a higher calculated dose within the screening‐
level risk assessment.
Chlorpyrifos is an organophosphate pesticide used in food
crop protection, and it inhibits the acetylcholinesterase enzyme
(Solomon et al., 2014). By disrupting cholinergic neuro-
transmission, chlorpyrifos and other organophosphates affect
physiological processes such as locomotion, grooming, and
odor learning in bees (Urlacher et al., 2016; Williamson et al.,
2013). In previous studies, chlorpyrifos was detected in pollen
(11–55 ppb), nectar (3.9 ppb), and honey (46 ppb) in crop fields
and suburban areas (Cutler et al., 2014; Mullin et al., 2010;
Ostiguy et al., 2019). Our reported values of chlorpyrifos de-
tected in pollen in all four states are consistent with the
wileyonlinelibrary.com/ETC
9
aforementioned values (Supporting Information, Table S2).
Our sole detection of chlorpyrifos in nectar (MI, 3.2 ppb,
Table 1) was also consistent with the value reported by Cutler
et al. (2014). In a fashion similar to imidacloprid, chlorpyrifos
was attributed a calculated RQ value of 0.1 for larvae reared in
vitro and exposed through food (Dai et al., 2019). Once again,
compared with our calculated RQ values for adults (0.62 in
nectar; Table 1), honey bee larvae could be at lower risk from
chlorpyrifos than adults if the compound is present in nectar.
The USDA NDL Gastonia Apiculture Pesticide Screen for
173 compounds (active ingredients and metabolites) of nectar
and pollen samples exhibit limits of detection ranging from 1 to
100 ppb. Although the number of compounds included in the
present study is substantial, it is not exhaustive, given there
were approximately 1300 registered pesticides in the United
States (including both synthetic and organic active ingredients)
at the time of the present publication. The LOD for a given
compound is also impacted by its chemical properties and the
matrix analyzed. As the LOD is improved, the frequency of
positive detections will likely increase in future similar studies.
Also, there is a limitation in LOD with multiresidue screenings
like QuECHeRS, which poses a trade‐off in detection sensitivity
as the number of compounds increases. Our data only per-
mitted us to address compounds that exceeded the LOD in the
screen. For compounds for which the LOD was less than the
LC50, the method we used is likely sufficient to estimate risk.
Nevertheless, the Apiculture Pesticide Screen was developed
to screen for compounds most likely to be a threat to honey
bees based on the frequency of use and the historic lack of
detection (Mullin et al., 2010), therefore weighting the analysis
to detect compounds historically perceived to be of im-
portance to honey bee health.
The extent to which our data can be extrapolated to non‐
Apis bees also needs to be evaluated. Honey bees are a rec-
ognized surrogate species for the USEPA pollinator risk as-
sessment framework, but uncertainties do exist regarding the
extent to which the honey bee risk assessment is protective for
other bee species (Cutler et al., 2014; Hinarejos et al., 2019).
Differences in exposure can occur through different exposure
routes (e.g., soil), consumption estimates, and floral visitations.
Whereas some non‐Apis bees are generalist pollinators, other
species, such as oligolectic bees, are restricted in their floral
visitations due to seasonality or floral source, which may either
reduce or increase their exposure to a given pesticide. In our
study, we analyzed residues of pollen and nectar that were
collected by honey bees, possibly excluding or limiting some
floral sources that are exclusively visited by non‐Apis bee
species. There is also limited information on the non‐Apis
species distribution in urban/suburban areas. During the first
year of collection, pollen samples were submitted for identi-
fication (Lau et al., 2019), and this can provide some per-
spective regarding exposure to non‐Apis species. However,
exposure to pesticides by non‐Apis bee species is an under-
studied area that could benefit from additional research efforts.
The current regulatory framework in the United States does
not consider pesticide mixtures when characterizing the polli-
nator risk associated with multiple compounds. Of the 328
© 2022 SETAC
10
pollen samples that had a positive detect (38.1% detection rate
out of 862 samples), 139 samples (42.1% of the positive de-
tects) had multiple pesticides detected. To a lesser extent, of
the 114 nectar samples that had a positive detect (14.8% of the
768 samples), 25 samples (21.9% of the positive detections)
had multiple pesticides detected. The potential for synergisms
among pesticides has been demonstrated in laboratory studies
in which some treatments for the parasitic mite Varroa de-
structor contributed to a greater‐than‐additive toxicity to honey
bees in combination with other compounds (Johnson et al.,
2006; Rinkevich et al., 2017). Our data do not exclude the
possibility of pesticide interactions, yet the frequency of syn-
ergistic interactions between two or more chemicals under
realistic environmental exposure scenarios is less common than
impacts from exposure to a single compound (Cedergreen,
2014; Levine & Borgert, 2018).
The topic of synergism has been researched extensively with
other taxa, but only more recently with honey bees. First, it is
important to define what constitutes synergy. Typically, it means
a deviation from an expected outcome assuming additive ef-
fects. The general consensus is that a minimum toxicity factor
of 2 to 5× is required for practical biological relevance in
an environmental risk assessment (Belden & Brain, 2018;
Cedergreen, 2014; EFSA, 2013b). From that approach, there are
available equations that can be used to estimate these model
deviation ratios, such as the Finney equation included in the
USEPA's Guidelines for the Health Risk Assessment of Chemical
Mixtures (1986). Many studies have been published on possible
synergistic impacts of pesticides on bees (Biddinger et al., 2013;
Gill et al., 2012; Iverson et al., 2019; Sgolastra et al., 2017;
Thompson et al., 2014; Zhu et al., 2017, among others). How-
ever, several of these studies (Gill et al., 2012; Zhu et al., 2017)
present results that can be characterized as additive effects, not
synergism (more than 5×). The other studies report results about
combinations of compounds that have long been known to have
synergistic effects. In most instances, proposed cases of synergy
can be explained mechanistically, and the exposure scenario can
be mitigated. There are some label restrictions in place when
true synergisms are known, aimed at limiting their co‐occurrence
in the environment. Of course, violations of the label could result
in off‐label product mixing, possibly leading to synergistic in-
teractions between compounds.
Our study is limited in characterizing how the frequency of
pesticide use translates to realistic compound residues in nectar
and pollen. In our study, CA is the state with the most stringent
regulation of pesticides, requiring an annual report of crop pro-
tection pesticides used to the Department of Pesticide Regu-
lation (the state's regulatory agency) through the Pesticide Use
Regulation program. However, even when some information on
nonagricultural uses of pesticides may be available for certain
counties, there are limitations to determining application rates
(e.g., amount of active ingredient used per hectare or per plant),
consumer products, plant species treated, method and time of
application, and bee visitation rates to the exposed nectar and
pollen matrices. Thus, it is not possible to associate pesticide use
rates with the resulting residue levels in the nectar and pollen
samples analyzed in our study.
© 2022 SETAC
Environmental Toxicology and Chemistry, 2022;00:1–13—F.J. Démares et al.
Our study provides critical information needed to charac-
terize pesticide exposure for honey bees in urban/suburban
landscapes, which have historically received little attention
(Kavanagh et al., 2021). We detected pesticide residues in 27%
(442/1630) of the pollen and nectar samples collected by for-
aging bees, but the vast majority of the pesticide residues
found in nectar and pollen posed minimal or no acute risk to
individual honey bees. Four pesticides did exceed the
screening level of concern for potential acute toxicity at the
maximum detected concentrations, although chronic risk
cannot be excluded, and may be greater, for the other com-
pounds detected. No major temporal or spatial patterns were
identified across the four sampling regions of FL, CA, MI, and
TX. All four compounds that exceeded the LOC are active in-
gredients in products approved for use in urban/suburban
settings. Thus, it is important that the products be applied per
label guidelines to minimize their impact on pollinators. Such
guidelines often include the prohibition of applying the prod-
ucts onto blooming plants. Ultimately, our study provides a
foundation for additional research on the major drivers of
honey bee colony mortality in nonagricultural landscapes.
Supporting Information—The Supporting Information is avail-
able on the Wiley Online Library at https://doi.org/10.1002/
etc.5298.
Acknowledgment—S. Luo helped with the Michigan sample
collection, and B. Stanford, S. Nease, S. Gamez, M. Bammer,
A. Wedde, B. Stanford, and R. Diaz (all University of Florida)
assisted with the Florida sample collection and preparation.
K. Kleckner and B. Johnson assisted with data curation. We
thank all beekeepers who allowed us to sample their colonies.
We also acknowledge the contributions of J. Barber and
R. Simonds of the National Science Laboratory, US Department
of Agriculture (USDA‐NSL), who conducted the multiresidue
analysis. Bayer CropScience and Syngenta Crop Protection
were the primary sponsors for the project through awards to
James D. Ellis, University of Florida (award 00115707), Juliana
Rangel, Texas A&M University (award M1402691), Zachary Y.
Huang, Michigan State University (award NSNTN015), and
Joseph Sullivan (Ardea Consulting). Additional sponsorship for
the University of Florida (to James D. Ellis) was provided by the
USDA National Institute of Food and Agriculture (Multi‐State
Hatch Project 1005822), Juliana Rangel's Texas AgriLife
Research Hatch Project (TEX09557), and a USDA National In-
stitute of Food and Agriculture Multistate Project (2015‐67013‐
23170). The USDA is an equal opportunity provider and
employer.
Disclaimer—Daniel Schmehl and Ana R. Cabrera are Bayer
CropScience employees.
Author Contributions Statement—James D. Ellis: Conception;
design of experiments; performance of experiments; statistical
analysis; writing—first draft. Daniel Schmehl: Conception;
design of experiments; performance of experiments; statistical
analysis; writing—first draft. Ana R. Cabrera: Conception;
wileyonlinelibrary.com/ETC
Honey bee exposure to pesticides in urban environments—Environmental Toxicology and Chemistry, 2022;00:1–13
design of experiments; statistical analysis; writing—first draft.
Zachary Y. Huang: Conception; design of experiments;
performance of experiments; writing—technical and editorial
assistance. Juliana Rangel: Conception; design of experi-
ments; performance of experiments; writing—technical and
editorial assistance. Joseph Sullivan: Conception; design of
experiments; performance of experiments; writing—technical
and editorial assistance. Pierre Lau: Performance of experi-
ments; writing—technical and editorial assistance. Xianbing
Xie: Performance of experiments; writing—technical and
editorial assistance. Fabien J. Démares: Statistical analysis;
writing—first draft. Jeffrey R. Bloomquist: Statistical analysis;
writing—first draft. All authors approved the final submitted
manuscript and accompanying materials.
Data Availability Statement—Data, associated metadata, and
calculation tools are available from the corresponding author
(jdellis@ufl.edu).
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