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Recent concern over global pollinator declines has led to considerable research on the effects of pesticides on bees. Although pesticides are typically not encountered at lethal levels in the field, there is growing evidence indicating that exposure to field-realistic levels can have sublethal effects on bees, affecting their foraging behaviour, homing ability and reproductive success. Bees are essential for the pollination of a wide variety of crops and the majority of wild flowering plants, but until now research on pesticide effects has been limited to direct effects on bees themselves and not on the pollination services they provide. Here we show the first evidence to our knowledge that pesticide exposure can reduce the pollination services bumblebees deliver to apples, a crop of global economic importance. Bumblebee colonies exposed to a neonicotinoid pesticide provided lower visitation rates to apple trees and collected pollen less often. Most importantly, these pesticide-exposed colonies produced apples containing fewer seeds, demonstrating a reduced delivery of pollination services. Our results also indicate that reduced pollination service delivery is not due to pesticide-induced changes in individual bee behaviour, but most likely due to effects at the colony level. These findings show that pesticide exposure can impair the ability of bees to provide pollination services, with important implications for both the sustained delivery of stable crop yields and the functioning of natural ecosystems.
548 | NATURE | VOL 528 | 24/31 DECEMBER 2015
© 2015 Macmillan Publishers Limited. All rights reserved
Neonicotinoid pesticide exposure impairs crop
pollination services provided by bumblebees
Dara A. Stanley
, Michael P. D. Garratt
, Jennifer B. Wickens
, Victoria J. Wickens
, Simon G. Potts
& Nigel E. Raine
Recent concern over global pollinator declines has led to
considerable research on the effects of pesticides on bees
Although pesticides are typically not encountered at lethal levels
in the field, there is growing evidence indicating that exposure to
field-realistic levels can have sublethal effects on bees, affecting
their foraging behaviour
, homing ability
and reproductive
. Bees are essential for the pollination of a wide variety of
crops and the majority of wild flowering plants
, but until now
research on pesticide effects has been limited to direct effects on
bees themselves and not on the pollination services they provide.
Here we show the first evidence to our knowledge that pesticide
exposure can reduce the pollination services bumblebees deliver to
apples, a crop of global economic importance. Bumblebee colonies
exposed to a neonicotinoid pesticide provided lower visitation rates
to apple trees and collected pollen less often. Most importantly,
these pesticide-exposed colonies produced apples containing fewer
seeds, demonstrating a reduced delivery of pollination services. Our
results also indicate that reduced pollination service delivery is not
due to pesticide-induced changes in individual bee behaviour, but
most likely due to effects at the colony level. These findings show
that pesticide exposure can impair the ability of bees to provide
pollination services, with important implications for both the
sustained delivery of stable crop yields and the functioning of
natural ecosystems.
Biotic pollination is required by a large proportion of crops world-
, disproportionately including those with economically high
values and nutritional content
. The contribution of pollination services
to global agriculture has been steadily increasing and was estimated at
US$361 billion in 2009 (ref. 14). In addition, animal-vectored pol-
lination is required by an estimated 87.5% of all angiosperms to
, making this process fundamental to the functioning of
natural ecosystems. Therefore, any threats to the delivery of pollina-
tion services could have serious consequences for both food security
and wider ecosystem function. Neonicotinoid pesticides, the most
widely used group of insecticides worldwide
, are implicated as one
of the contributing factors in the global declines of bee pollinators
Although previous work has shown that bumblebee foraging activity,
colony growth and reproduction can be altered by sublethal exposure
to neonicotinoid pesticides
, all research on pesticide effects has
focused on bees as the service providers, but has not assessed the polli-
nation service itself. Therefore it is unknown whether pesticide exposure
actually results in changes to the delivery of pollination services to crops
and wild plants (for a discussion of potential mechanisms see ref. 17).
This information is essential to assess the severity of pesticide effects on
ecosystem services, and to inform actions to mitigate negative effects.
Apples are an important global crop, with 75 million tonnes har-
vested from 95 countries in 2012 and an estimated export value of
US$71 billion (Food and Agriculture Organisation statistics, http:// Apple crops benefit from insect pollination with
seed number, fruit set, fruit size and shape all improved with increased
pollination services
. Bumblebees are major pollinators of apples
and many other crops across the world
, and are exposed to low levels
of pesticides when foraging in agricultural areas. Here we investigated
how exposure to low, field-realistic levels of a widely used neonicoti-
noid insecticide (thiamethoxam) could affect the ability of bumblebees
to pollinate apple trees. We pre-exposed colonies to 2.4 parts per billion
(ppb) thiamethoxam, 10 ppb thiamethoxam or control solutions (con-
taining no pesticide; rationale for selecting pesticide concentrations
and relevance of results are outlined in Methods and Supplementary
Information) in their nectar source (artificial sugar water) for a period
of 13 days (8 colonies per treatment, that is, 24 colonies in total).
Subsequently, colonies were brought to the field and allowed access to
virgin apple trees of a dessert (Scrumptious) variety, along with trees
of a polliniser (Everest) variety, in pollinator exclusion cages in which
we observed both individual- and colony-level behaviour. At the end of
the season, apples from tested trees were collected to assess pollination
service delivery in terms of fruit and seed set.
When whole colonies were given access to apple trees we found
an effect of insecticide treatment on visitation rates to apple flowers
= 3.1, P = 0.05); colonies exposed to 10 ppb pesticide provided
lower visitation rates to apple flowers than controls (Fig. 1a; Extended
Data Table 1). We also found an effect of treatment on the number of
foraging trips from which bees returned carrying pollen (χ
= 9.65,
degrees of freedom (df) = 2, P = 0.008), with fewer bees from colo-
nies exposed to 10 ppb pesticide returning with pollen than work-
ers from control colonies (Fig. 1b). Apple abortion rate was affected
by treatment (χ
= 5.94, df = 2, P = 0.05), with trees pollinated by
School of Biological Sciences, Royal Holloway University of London, Egham TW20 0EX, UK.
Centre for Agri-Environmental Research, School of Agriculture, Policy and Development, University of
Reading, Reading RG6 6AR, UK.
School of Environmental Sciences, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Control 2.4 ppb 10 ppb
Visitation rate per patch
(no. visits per ower per min)
No. bees carrying pollen
Control 2.4 ppb 10 ppb
Figure 1 | Effects of pesticide treatment on colony-level behaviour.
a, b, Visitation rates provided by colonies to Scrumptious apple flowers
(number of visits per flower per minute) (a) and number of foraging trips
from which bees returned carrying pollen (b), from colonies exposed to
different pesticide treatments. Eight colonies were observed per treatment
group, and means ± s.e.m. are shown, *P < 0.05. NS, not significant.
Results from statistical models are given in Extended Data Table 1.
24/31 DECEMBER 2015 | VOL 528 | NATURE | 549
© 2015 Macmillan Publishers Limited. All rights reserved
2.4 ppb pesticide-exposed colonies aborting more fruit than controls
(Fig. 2a), although overall levels of fruit set did not differ (χ
= 4.1,
df = 2, P = 0.13) and there was no difference in the proportion of trees
that produced fruit among treatments (χ
= 1.2, df = 2, P = 0.55).
However, we found a significant effect of treatment on the number
of seeds produced per apple, an indicator of fruit quality, (χ
= 8.27,
df = 2, P = 0.02); flowers pollinated by colonies exposed to 10 ppb pesti
cide produced significantly fewer seeds than those pollinated by 2.4 ppb
colonies (Fig. 2b). These results show that colonies exposed to pesticide
can deliver reduced pollination services to apple crops.
These colony-level effects could be explained by several mecha-
nisms, including individual behavioural changes. Individual bees
exposed to 10 ppb pesticide spent longer foraging (F
= 3.72, P = 0.03;
Fig. 3a), visited more Scrumptious flowers (χ
= 12.79, df = 2,
P = 0.002) and switched more frequently between varieties dur
ing each trip (χ
= 11.32, df = 2, P = 0.003: Fig. 3b; Extended Data
Table 2), which suggests a modification of their floral preferences
Neonicotinoids target neurotransmitter receptors in insects and, as
well as causing neuronal inactivation
, some have been shown to be
partial neuronal agonists
; therefore increases in individual foraging
activity may be explained by acute increases in neuronal activity caus-
ing hormesis (a biphasic response in which low levels of an otherwise
toxic compound can result in stimulation of a biological process
However, we found no effect of treatment on whether flowers visited
by these individual bees produced apples (χ
= 0.88, df = 2, P = 0.64),
showed higher rates of fruit abortion (χ
= 0.42, df = 2, P = 0.81) or
different levels of seed set (χ
= 0.11, df = 2, P = 0.95). This suggests
that bees exposed to pesticide must somehow be behaving differently
on flowers, in a way that was not readily observable in our experiment
(for example, changes in stigmatic contact
), such that increased visit
frequency did not result in better pollination service delivery at the
individual level.
Our results suggest that effects on pollination service delivery are
not due to individual behavioural modification, but instead are most
likely due to changes in colony activity levels as evidenced by reduced
floral visitation rates and pollen collection. Bees collecting pollen may
be more effective pollinators as they can deposit more pollen on plant
; therefore if pesticide-exposed colonies are collecting less
pollen they are also likely to be depositing less on stigmas than bees
from control colonies. While individual bees exposed to pesticides
visited more flowers, overall pesticide-exposed colonies provided lower
visitation rates and collected less pollen, thus explaining why reduced
pollination services were delivered. Gill & Raine
found that control
(untreated) bees improved their pollen foraging performance over time,
whereas imidacloprid-treated bees became less successful foragers;
foragers in our colony-level experiment may have carried out multiple
trips and become more experienced foragers, potentially explaining
why we find effects on pollen collection here but not in the individual-
level experiment. Interestingly, for almost all parameters measured in
this study we found significant effects on both individual behaviour
and colony-level function following 10 ppb thiamethoxam exposure,
but not at the 2.4 ppb level. This suggests that there are dose-dependent
effects that lie between these two exposure levels. Both these exposure
levels are highly relevant as they are within the range measured in the
field, but further work is necessary to elucidate the lowest level at which
these effects become significant (for further discussion of rationale for
exposure and relevance of results, see Methods and Supplementary
A 36% reduction in the number of seeds produced in apples polli-
nated by colonies exposed to 10 ppb pesticide in comparison to control
colonies has important agronomic implications for crop production.
The number of seeds in apples is closely linked to fruit crop quality in
most, but not all, varieties
and the enhancement of fruit quality,
particularly the proportion of Class 1 fruit, underpins the economic
value of UK orchards
: growers must typically thin out their apple
crops making the quality of each fruit very important. Therefore
impacts on seed set and fruit quality have direct implications for apple
production value, and as seed set and fruit set are positively linked
in many varieties, reduced seed set can have direct negative implica-
tions for fruit set and total crop yield
. As certain apple varieties in
the UK currently experience pollination deficits
, mitigating the
effects of pesticides on bumblebee pollinators could improve polli-
nation service delivery. Apple crops are visited by a wide variety of
pollinator groups, and neonicotinoid pesticides differentially affect
insect taxa
. Apart from bumblebees, one of the other main polli-
nator groups that visit apple flowers are solitary bees
, and it has been
suggested that pesticide sensitivity of solitary bees is likely to be higher
than for larger, social species like bumblebees
. Therefore, apple
pollination in a field setting could be more vulnerable to pesticide
exposure than measured here.
Bumblebees are essential pollinators of many important crops other
than apples, including field beans, berries, tomatoes and oilseed rape
If exposure to pesticides alters pollination services to apple crops, it
is likely that these other bee-pollinated crops would also be affected.
Most importantly, the majority of wild plant species benefit from insect
Proportion of fruit set
2.4 ppb
10 ppb
Control 2.4 ppb 10 ppb
No. seeds produced per apple
Figure 2 | Effects of pesticide treatment on fruit and seed set.
a, b, The change in proportion of fruit set for trees (48 trees in total, 16 per
treatment) pollinated by colonies exposed to different pesticide treatments
measured early (May) and late (September), which represents fruit
abortion level (a), and number of seeds produced per apple (134 apples in
total; 53 in control, 46 in 2.4 ppb and 35 in 10 ppb pesticide treatments)
pollinated by colonies exposed to different pesticide treatments (b). Eight
colonies were observed per treatment group, and means ± s.e.m. are
shown, *P < 0.05, † indicates a difference of P = 0.06 between control and
10 ppb. NS, not significant. Results from statistical models are given in
Extended Data Table 1.
Control 2.4 ppb 10 ppb
Length of time spent foraging (s)
No. switches between varieties
Control 2.4 ppb 10 ppb
Figure 3 | Effects of pesticide treatment on individual bee behaviour.
a, b, Time spent foraging per foraging trip (seconds; n = 68 bees) (a) and
number of switches between Scrumptious and Everest apple varieties
(n = 93 bees) (b) for individual bees exposed to different pesticide
treatments. Means ± s.e.m. are shown, *P < 0.05, † indicates a difference
of P = 0.06 between control and 2.4 ppb. NS, not significant. Results from
statistical models are given in Extended Data Table 2.
550 | NATURE | VOL 528 | 24/31 DECEMBER 2015
© 2015 Macmillan Publishers Limited. All rights reserved
pollination services
. Therefore reduced pollination by pesticide-
affected colonies, as evidenced by reduced seed set, also has significant
implications for pollination in wild systems. Many wild plant species
are both self-incompatible and pollen limited
, so any reduction in the
delivery of pollination services could have substantial effects on wild
plant communities and therefore wider ecosystem function.
Concerns over global bee declines are strongly driven by the need for
the essential pollination services they provide to both crops and wild
plants. The use of neonicotinoid pesticides presents a potential threat to
bee health and, although the evidence base reporting sublethal (behav-
ioural) effects of pesticides on bees is mounting
, we have shown for the
first time that there is also an important effect of pesticide exposure on
the pollination services bees provide. This information provides a new
perspective when trying to fully understand the trade-offs involved
when using insecticides, showing that both the potential benefits and
the true costs of pest control options need to be considered.
Online Content Methods, along with any additional Extended Data display items and
Source Data, are available in the online version of the paper; references unique to
these sections appear only in the online paper.
Received 8 July; accepted 23 October 2015.
Published online 18 November 2015.
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Supplementary Information is available in the online version of the paper.
Acknowledgements We thank technicians at the University of Reading for
assistance in apple collection and seed counting, and E. van Leeuwen and
colleagues at Royal Holloway University of London for useful discussions. This
study was supported by UK Insect Pollinators Initiative grants BB/I000178/1
awarded to N.E.R. and BB/1000348/1 awarded to S.G.P. (funded jointly by the
Living with Environmental Change programme, Biotechnology and Biological
Sciences Research Council (BBSRC), Wellcome Trust, Scottish Government,
Department for Environment, Food and Rural Affairs (Defra) and Natural
Environment Research Council (NERC)). N.E.R. is supported as the Rebanks
Family Chair in Pollinator Conservation by The W. Garfield Weston Foundation.
Author Contributions D.A.S. and N.E.R. conceived the project, D.A.S., N.E.R. and
M.P.D.G. designed the research, D.A.S., J.B.W and V.J.W. carried out the research,
D.A.S., N.E.R., M.P.D.G. and S.G.P. contributed equipment for the research, D.A.S.
analysed the data, all authors were involved in writing the manuscript.
Author Information Reprints and permissions information is available at The authors declare no competing financial
interests. Readers are welcome to comment on the online version of the
paper. Correspondence and requests for materials should be addressed to
D.A.S. ( or N.E.R. (
© 2015 Macmillan Publishers Limited. All rights reserved
Pesticide preparation. A stock pesticide solution was made by dissolving 100 mg
thiamethoxam (PESTANAL, Analytical Standard, Sigma Aldrich) in 100 ml
acetone (1 mg ml
). Aliquots of stock solution were added to 40% sucrose to
create treatment solutions of 10 μg l
(10 ppb) and 2.4 μg l
(2.4 ppb) thiameth-
oxam. These concentrations were chosen as field-realistic; the lower concentration
(2.4 ppb) was based on thiamethoxam concentrations found in nectar pots of
bumblebee colonies foraging in agricultural areas in the UK
and in pollen
collected by honeybees
, and the higher concentration (10 ppb) is within the range
measured in pollen and nectar and of a variety of treated crops
and contam-
inated wild flowers
, and has been used in previous studies examining effects
of another neonicotinoid (imidacloprid) on bumblebee behaviour
. A control
solution was also made by repeating the process outlined above but using an aliquot
of 10 ppb acetone only (that is, no pesticide).
Experimental setup. Twenty-four commercially reared Bombus terrestris audax
colonies were obtained from Biobest (Westerlo, Belgium) at the start of April 2014,
each containing a queen and an average of 99 workers (range 57–133).
Colonies were weighed on arrival to estimate the overall colony size, and each
assigned sequentially to one of three treatment groups (2.4 ppb thiamethoxam,
10 ppb thiamethoxam and control) based on decreasing mass (but randomly
assigned within block). Each day, three colonies (one from each treatment)
were assigned to treatment groups, until after 7 days all colonies were receiv-
ing treated sucrose (16 colonies exposed to thiamethoxam and 8 to control
solution). We chose this sequential exposure regime to mimic subsequent
field testing and ensure all colonies had comparable durations of exposure
to their treatment. Colonies were fed treated sucrose solution from a gravity
feeder inserted at the base of the nest box. Feeders were initially refilled every
2–3 days, and then every 1–2 days when the colonies had grown significantly.
Untreated, defrosted honeybee-collected pollen was provided to colonies every
2–3 days. Colonies were exposed to treatments for an average of 13 days (range
12–15) before field testing. Before being moved to the field, colonies had access
to a feeder containing sucrose (40%) in a laboratory flight arena for 48 h to
become accustomed to leaving the nest to forage. There was no difference in
colony weights at the start (ANOVA: F
= 0.091, P = 0.91) or end (ANOVA:
= 0.88, P = 0.43) of the experimental period, indicating no treatment effect
on colony size.
Field testing. Cage experiments were carried out at Sonning Farm, University
of Reading, UK. 100 apple trees of a commercial dessert apple (Scrumptious
variety) were moved into holding pollinator exclusion cages in mid-March 2014
before flowering to prevent insect visitation. Field experiments began when
trees were entering full flower in mid-April. Each day, one colony from each
treatment was taken from the laboratory, placed individually in one of the
three test cages and observed simultaneously (with one observer per cage) in
a randomized block design (see below for details of observations). Each day a
different treatment was assigned to each observer. Cages were 4.8 × 2.1 × 2.1 m
frames covered in polyethylene mesh (gauge size = 1.33 mm, Extended Data
Fig. 1). Observations were carried out on 8 dry, bright days from 16–26
April 2014 spanning the peak flowering of apples (daily means: maximum
temperature 16 °C, rainfall 2.5 mm). This flowering period limited the number of
days on which testing could be carried out, and therefore the number of colonies
that could be tested; as a result no statistical methods were used to predetermine
sample size. The investigators were not blinded to allocation during experiments
and outcome assessment.
Individual-level measurements. Each morning, three cages were pop-
ulated with two virgin Scrumptious trees each from the holding cages
(mean ± s.e.m. = 130 ± 8.5 flowers per tree) as well as two polliniser trees (Everest
variety, mean ± s.e.m. = 305 ± 15 flowers per tree, Extended Data Fig. 1). The
number of flowers of each variety was standardized across cages to ensure equal
floral density each day, and 40 open and receptive flowers were marked with
cable ties on each Scrumptious tree for subsequent estimation of pollination ser-
vices (fewer flowers were marked on the last day of observations as there were
no longer 40 full-bloom flowers—flower numbers on these days were noted).
The nest boxes in each cage were then opened to allow a single worker to exit.
This bee was observed for the duration of its foraging trip (until it attempted
to return to the nest), or until 60 min had elapsed (Extended Data Fig. 2). The
duration of the foraging trip, the number of flowers of each apple variety visited,
and the handling time for each flower visit was recorded using Etholog software
(EthoLog: Behavioural observation transcription tool, University of Sao Paulo,
Brazil, 2011). If the individual bee did not visit any flowers within the first 20 min,
it was assumed not to be a forager and was captured, returned to the colony and
another bee released. All bees that foraged were paint-marked before they were
returned to the colony to ensure the same individuals were not observed twice.
This process was repeated until all cages had the same number of active foragers
recorded (3–5 bees per colony each day). Individual level observations took place
between 10:00 and 16:30.
Colony-level measurements. After individual-level observations, the two focal
Scrumptious trees in each cage were removed and replaced with two new virgin
trees. Again we standardized the number of flowers of each variety across cages
with 40 open and receptive flowers on each tree marked with cable ties. Colony
boxes were opened to allow free entry and exit to all active bees for a period of
60 min. This time period was chosen to avoid over-pollination of test flowers based
on pilot observations. Colony activity was monitored at the nest entrance using
video cameras. After an initial 10-min period to allow the bees to become accus-
tomed to the setup, four 10-min focal observations were carried out on separate
patches of Scrumptious flowers in each cage to estimate visitation rates. At the
end of the 60-min period, the Scrumptious trees were immediately removed to
prevent further visitation. Colony level observations were carried out between
14:30 and 18:30.
Estimation of pollination services. At the end of both the individual and colony
observation periods, all test trees were returned to holding cages in which they
were not visited by any other insects until apples were harvested at the end of the
season. An initial assessment of fruit set from marked flowers (indicating flowers
open during cage tests) was made at the end of May for all test Scrumptious trees to
assess how many flowers were proceeding to fruit set stage (and how many aborted,
Fig. 2a). Marked apples were collected on 27 August, and a final assessment made of
the proportion of marked flowers that had produced mature fruit (Extended Data
Fig. 2). In the lab, seed number was counted per apple for all collected fruit (274
apples from 96 trees across both experiments). Details of all data analyses carried
out are given in the supplementary information.
Data analysis. Individual level. Measures of the number of flowers visited,
numbers of switches between apple varieties, duration of total time in cage
(from when the bee left the colony box until it returned/end of 60 min period)
and time taken to visit the first flower (latency) were recorded for all indi-
vidual bees. For 68 of 93 bees observed (evenly distributed across cages and
treatments) a number of additional response variables were also recorded
including mean duration of the first 5 flower visits, number of inter flower
intervals longer than 60 s, mean duration of flower visits, mean period of time
between flower visits, length of time spent foraging (time between first and last
flower visit) and total time spent on flowers (sum of durations for all individual
flower visits). We tested for differences in these measures among treatments by
constructing mixed-effects models with pesticide treatment as a fixed effect.
As several variables differed among days, including weather, floral abundance
and the identity of colonies used, day of testing was included as a random
blocking factor in all models. Data were analysed in R version 3.1.0 (ref. 38),
using either linear mixed effects (LME) models with the lmer function in the
nlme package for continuous data
, generalized mixed effects (GLMM) models
with Poisson distribution used for response variables that were counts using the
glmer function in the lme4 package
, or the glmmPQL function in the MASS
when data were overdispersed. Models were validated by plotting
standardized residuals versus fitted values, normal qq-plots and histograms of
residuals, and continuous response variables were logarithmically transformed
(log (X + 1)) if necessary to improve residual fit. If treatment was significant,
Tukey’s post hoc tests were performed using the glht function in the multcomp
To assess differences in apple production on trees visited by pesticide exposed
and control bees, we examined a number of variables including the number
of fruits produced at the start of the season (May) and at the end (September;
Fig. 2a), the change in proportion of apples forming from marked flowers per tree
between the start and end measures (fruit abortion levels) and number of seeds
per apple (measured in early September; Fig. 2b). Models were run as described
previously with treatment as a fixed effect, although the tree on which fruits
were produced, the number of bees released and date of testing were included
as random effects. As a number of trees produced no fruit, seed set data were
analysed in two steps. First, we tested whether there was a treatment difference
in the number of trees that produced any fruit. Second, we tested for treatment
differences in seeds per apple (a measure that only included trees that had
produced some fruit).
Colony level. We tested for differences in colony activity levels (the combined
number of entries and exits by workers to the colony box) and the number
of bees carrying pollen among treatments using GLMM models in the MASS
, with Poisson distribution for count data. Treatment differences in
flower visitation rate to Scrumptious trees were tested using LME models
. Date
of testing was used as a random effect in all models (and patch included as a
random effect in the flower visitation rate model), and models were validated as
© 2015 Macmillan Publishers Limited. All rights reserved
described above. Fruit abortion and seed set variables were analysed as described
for the individual level experiment, using tree and date of testing as random
31. Thompson, H. et al. Eects of neonicotinoid seed treatments on bumble bee
colonies under eld conditions. (Food and Environment Research Agency
(FERA), 2013).
32. Pilling, E., Campbell, P., Coulson, M., Ruddle, N. & Tornier, I. A four-year eld
program investigating long-term eects of repeated exposure of honey bee
colonies to owering crops treated with thiamethoxam. PLoS One 8, e77193
33. Castle, S. J., Byrne, F. J., Bi, J. L. & Toscano, N. C. Spatial and temporal
distribution of imidacloprid and thiamethoxam in citrus and impact on
Homalodisca coagulata populations. Pest Manag. Sci. 61, 75–84 (2005).
34. Dively, G. P. & Kamel, A. Insecticide residues in pollen and nectar of a cucurbit
crop and their potential exposure to pollinators. J. Agric. Food Chem. 60,
4449–4456 (2012).
35. Botías, C. et al. Neonicotinoid residues in wildowers, a potential route of
chronic exposure for bees. Environ. Sci. Technol. 9, 12731–12740 (2015).
36. Krupke, C. H., Hunt, G. J., Eitzer, B. D., Andino, G. & Given, K. Multiple routes of
pesticide exposure for honey bees living near agricultural elds. PLoS One 7,
e29268 (2012).
37. Stewart, S. D. et al. Potential exposure of pollinators to neonicotinoid
insecticides from the use of insecticide seed treatments in the
mid-southern United States. Environ. Sci. Technol. 48, 9762–9769
38. R Development Core Team. R: A language and environment for statistical
computing. R Foundation for Statistical Computing, Vienna, Austria. (2011).
39. Pinheiro, J., Bates, D., DebRoy, S., Sarkar, D. & R Development Core Team.
Package “nlme”: Linear and nonlinear mixed eects models. R package
version 3.1-104 (2012).
40. Bates, D., Maechler, M., Bolker, B. & Walker, S. lme4: Linear mixed-eects
models using Eigen and S4. R package version 1.1-7 http://CRAN.R-project.
org/package= lme4 (2014).
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(Springer, 2002).
42. Hothorn, T., Bretz, F. & Westfall, P. Simultaneous inference in general
parametric models. Biom. J. 50, 346–363 (2008).
© 2015 Macmillan Publishers Limited. All rights reserved
Extended Data Figure 1 | An example of the experimental setup at the Sonning Farm field site. Experimental pollinator exclusion cages containing a
bumblebee colony (located in the corner of the cage) and potted experimental apple trees are shown. Photos: D.A.S.
© 2015 Macmillan Publishers Limited. All rights reserved
Extended Data Figure 2 | An experimental bumblebee (Bombus terrestris) worker visiting an apple flower (left), and an example of an apple
produced from a marked (yellow cable tie) apple flower (right; Scrumptious variety). Photos: D.A.S. and C. L. Truslove.
© 2015 Macmillan Publishers Limited. All rights reserved
Extended Data Table 1 | Results from the colony-level experiment
Signicant dierences (P 0.05) are highlighted in bold.
© 2015 Macmillan Publishers Limited. All rights reserved
Extended Data Table 2 | Results from the individual-level experiment
Signicance dierences (P 0.05) are highlighted in bold.
... However, in this scenario, honey bees and other beneficial insects can be exposed to residual pesticides by gathering pollen or nectar, causing "colony collapse disorder" (Cox-Foster et al. 2007;Zhang et al. 2020a) and resulting in bee damage or death. It is certainly alarming that the pollination pathways of crops will be affected by NNI application, which may prevent crops from normal growing (Stanley et al. 2015). Given the potential health impacts and ecological risks from NNI usage, the European Union (EU) has completely banned the outdoor usage of IMI, THM, and CLO since 2018 (EU 2018a(EU , 2018b(EU , 2018c. ...
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Neonicotinoid insecticides (NNIs) have been widely used to control insect pests, while their environmental residues and associated hazardous impacts on human and ecosystem health have attracted increasing attention worldwide. In this study, we examined the current levels and associated spatial and temporal patterns of NNIs in multiple environmental media across China. Concentrations of NNIs in surface water, sediment, and soil were in the range of 9.94–755 ng·L⁻¹, 0.07–8.30 ng·g⁻¹ DW, and 0.009–356 ng·g⁻¹ DW, respectively. The high levels of NNIs in surface water, such as in Yangtze River (755 ng·L⁻¹), North River (539 ng·L⁻¹), Nandu River (519 ng·L⁻¹), and Minjiang River (514 ng·L⁻¹), were dominated by imidacloprid, thiamethoxam, and acetamiprid due to their extensive use. The levels of NNIs in sediments were relatively low, and the highest concentration (8.30 ng·g⁻¹ DW) was observed in Dongguan ditch. Sediment–water exchange calculated from fugacity fraction indicated that NNIs in sediment can be released back into the water due to their high solubility and low KOW. Soils from agricultural zones contained the largest residual NNIs, with imidacloprid concentrations in cultivated soil reaching 119 ng·g⁻¹ DW. The calculated leaching potential showed that clothianidin has the highest migration potential to deep soil or groundwater. The monitored data of NNIs presented a decreasing trend from 2016 to 2018, which might be caused by the implementation of relevant control policies for NNI applications. The high levels of NNIs mainly occurred in southern China due to frequent agricultural activities and warm and humid meteorological conditions. The results from this study improve our understanding of the pollution levels and environmental behavior of NNIs in different environmental media across China and provide new knowledge that is needed for making future control policies for NNIs production and application. Graphical abstract
... In the last decades, a concern about a global decline of pollinators has resulted in an increasing focus on the effect of pesticides on honeybees (Apis mellifera sp.) and wild bees (Faita et al., 2018;Lundin et al., 2015;Mulvey & Cresswell, 2020;Stanley et al., 2015). Also, pesticide residues in nectar and pollen in flowers (Bonmatin et al., 2015;Botias et al., 2015;Gierer et al., 2019), and in the pollen and honey collected by the bees have attracted more attention (El-Nahhal, 2020;Mitchell et al., 2017;Mullin et al., 2010;Raimets et al., 2020;Simon-Delso et al., 2017). ...
Clopyralid is a systemic herbicide used in oilseed rape and other crops. It was found in Danish honey from 2026 in concentrations exceeding the maximum residue level (MRL) of 0.05 mg kg⁻¹. About 50% of the Danish honey is based on nectar from winter oilseed rape. In 2019 and 2020, winter oilseed rape fields were sprayed with clopyralid just before the assigned spraying deadline. At flowering, nectar and pollen samples were collected and the content of clopyralid was measured. Honey and pollen samples were also collected from beehives next to ten conventional winter oilseed rape fields sprayed with clopyralid. Clopyralid was found in nectar and pollen from the experimental fields, and in honey and pollen from beehives next to the conventional fields. For most samples the content in nectar and honey exceeded the MRL. The concentrations found, may not pose any health risk for consumers, as the MRL is based on the original detection limit and not on toxicological tests. However, it can have a significant economically consequence for the beekeepers, who are not allowed to sell the honey if the concentration of clopyralid exceeds 0.1 mg kg⁻¹. Reducing the acceptable applicable rate of clopyralid or implement an earlier deadline for spraying of clopyralid may reduce the risk of contaminating bee food products. However, if it is not possible to obtain a satisfactory effect of clopyralid on the weed flora under these conditions, spraying with pesticides containing clopyralid should be restricted in winter oilseed rape. Determination of an MRL value based on toxicological tests might result in a higher value and makes it acceptable selling the honey containing higher levels of clopyralid.
... To date, intensive agricultural landscapes have been increasingly simplified and pests are mostly controlled with pesticides. But, pesticides not only may prove to be increasingly ineffective and environmentally detrimental, but in addition they may also impact non-target organisms (Iyaniwura 1991;Pelosi et al. 2014;Zaller and Brühl 2019) and thus impede ecological functioning in such treated agroecosystems (Stanley et al. 2015;Prashar and Shah 2016). This raises several questions. ...
Central to the agroecological transition is biodiversity, which, once restored in agroecosystems, should provide multiple ecosystem services that result from the interactions between different organisms. However, the diversity of expected services raises questions about the capacity of agroecosystems to simultaneously provide many of them as well as the possible existence of synergies and antagonisms. In particular, the relationships between crop-associated biodiversity (i.e. biodiversity in action in crops) and spontaneous biodiversity (i.e. biodiversity not directly related to agriculture) remain unclear. In this article, we analyse the impact of hedgerows on the preservation of spontaneous biodiversity, on the promotion of biotic regulation services such as pollination and pest control in agriculture, and on the interactions between spontaneous and associated biodiversity. Our analysis of the scientific literature shows that hedgerows are unique assets for the preservation of spontaneous biodiversity, while they also provide biotic regulation services to adjacent crops but only under specific conditions, which need to be better understood. We propose a functional conceptual framework of the ecological effects of hedgerows on associated biodiversity and we highlight the possible synergistic and antagonistic effects related to hedgerow characteristics and the life-history traits of the organisms under consideration. Our analysis therefore seeks to overcome the cleavage between associated biodiversity and spontaneous biodiversity. This approach is in keeping with a more harmonious relationship between agriculture and biodiversity.
... In recent years, the negative effects of neonicotinoids in organisms have been studied, including their associated underlying risk to human health (Hallmann et al. 2014). It has been reported that neonicotinoids reduce the reproduction of bees and increase their mortality by inhibiting their homing ability (Rundlof et al. 2015;Stanley et al. 2015;Woodcock et al. 2017). The metabolism and dissipation fate of IMI and ACE on are relatively safe. ...
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Mixed pesticides have been broadly used in agriculture. However, assessing the combined effects of pesticides in the environment is essential for potential risk assessment, though the task is far from complete. Median lethal concentrations of pesticides as well as acetylcholinesterase (AChE) levels and cellulose activities were measured in earthworms (Eisenia fetida) individually and jointly exposed to pesticides imidacloprid (IMI), acetamiprid (ACE), chlorpyrifos (CRF), and abamectin (ABM)). A 3:1 mixture of CRF and IMI had additive effects, while a 3:1 mixture of CRF and ACE had synergic effects. The joint effects of ABM with IMI or with ACE were synergistic. As CRF concentration increased, AChE activities were significantly decreased. For high concentrations of IMI, AChE activities under combined CRF and IMI applications were significantly inhibited following increased exposure time. Moreover, the cellulase activities under combined applications of CRF with IMI or with ACE had similar effects. This study provides basic data for scientifically evaluating the environmental risk and safety of combined uses of pesticides. Graphical abstract
... There are a number of environmental factors that have been found to negatively affect pollinator populations worldwide: the most important stressors include the lack of a variety of food sources due to intensive agricultural production (Naug 2009;Donkersley et al. 2014), infection by parasites and diseases Cameron and Sadd 2020), extreme weather due to climate change (Kerr et al. 2015;Martinet et al. 2020), and exposure to various pesticides (Blacquière et al. 2012;Botías et al. 2021). As the current trends in pollinator decline have the potential to jeopardize future sustainability of crop pollination in many agroecosystems (Stanley et al. 2015;Whitehorn et al. 2017;Fijen et al. 2018), it has become essential to explore the pathways through which such environmental factors interfere with pollinator life histories and take actions to mitigate their detrimental effects. ...
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Due to recent changes in regulation, acetamiprid has become the only neonicotinoid that can be applied without restrictions and in open field cultivations in the EU from 2021. We provide an overview of the current knowledge on the effects of this insecticide on bumblebees and assessed whether available empirical evidence supports the claim that acetamiprid poses negligible risk to these pollinators. We found that there is limited data on the lethal and sublethal effects of this pesticide on bumblebees. While risk assessment results suggest that field-realistic concentrations of acetamiprid have minor acute and chronic toxicity, detrimental sublethal effects, including reduced reproductive output, have been observed when bumblebees were exposed to high doses of this insecticide. We propose that further research on the topic is warranted as the more extensive application of acetamiprid may lead to such high concentrations in the field.
... To meet the increasing food demand and effectively control rice diseases, insect pests and weeds, intensive agricultural management with monoculture, high-yield varieties and injudicious use of agrochemicals has been widely adopted since the Green Revolution (Pimentel, 1996;Saha & Schmalzer, 2016). However, these measures resulted in pesticide resistance in addition to the adverse impacts on non-target organisms and ecosystem services (Jin et al., 2017;Matsumura et al., 2010;Stanley et al., 2015), hindering sustainable food production (Lu et al., 2015;Veleva & Ellenbecker, 2001). Therefore, with people's concern about food security, there is a growing need to reduce the use of chemicals in the food production systems (Mac Loughlin et al., 2018;Wang et al., 2009;Zhu et al., 2017). ...
This study was conducted to evaluate whether conventional farming can be replaced by a lower chemical pollution farming system, integrated rice-duck farming in a suburban agricultural area in northern Zhejiang, Southeast China. We comprehensively assessed the impacts of integrated rice-duck farming on weeds and herbivorous insects along with their natural enemies during the rice-growing season. We found that the rice-duck system effectively controlled weeds within the paddy fields. Nevertheless, the abundance of herbivores, particularly planthoppers and leafhoppers was increased in the duck-roaming paddy fields compared to the control and conventional fields, indicating the inefficiency of the rice-duck system in the control of rice main herbivore pests. Rice-duck farming significantly reduced the abundance of predators, especially spiders and dragonflies, therefore an ‘enemy-free space’ was created for herbivores which might be responsible for the observed herbivore increase in the rice-duck fields. Despite the inefficiency of rice-duck farming in the control of rice main herbivore pests, the net income of the rice-duck treatment with an organic certification was 7.6 and 1.4 times higher compared to the conventional and control treatments, respectively. In the view of the favourable results obtained in this study, the integrated rice-duck farming system might be suggested as a high economic return organic farming method suitable for modern farmers.
A major challenge in sustainable agriculture is finding solutions to manage crop-damaging pests such as herbivores while protecting beneficial organisms such as pollinators. Squash is a highly pollinator-dependent crop that is also attractive to herbivores like the striped cucumber beetle. While synthetic insecticides can provide control of insect pests, they can also affect non-target organisms such as pollinators. Thus, growers need to balance pest management with pollinator protection to ensure optimal yield. Thiamethoxam is a commonly used systemic insecticide that translocates throughout plants, leaving residues in nectar and pollen. The aim of this study was to evaluate whether there are uses of this insecticide that provides efficient pest control while minimizing pesticide pollinator exposure. Specifically, we tested how different prophylactic application methods (seed treatments, in-furrow applications, and early foliar sprays) of commercially available thiamethoxam products impact pest control, bee visitation, yield, and pesticide residues in flowers of squash crops. We found that among the different methods of thiamethoxam application, in-furrow application best prevented defoliation and resulted in the highest fruit weight and number. However, it also produced the most frequent and highest concentrations of thiamethoxam in nectar and pollen, reaching lethal levels for squash bees. Our study provides evidence that under current application methods, thiamethoxam does not provide a sustainable solution for squash growers and further research is required on more efficient pesticide delivery methods, as well as non-pesticide pest control measurements.
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Hoverflies (Diptera: Syrphidae) and bees (Hymenoptera: Anthophila) are two key taxa for plant pollination. In the present research, the altitudinal distribution of these taxa was studied along two gradients (elevation range: 780–2130 m) in the Dolomiti Bellunesi National Park (Northeastern Italy). Pan traps were used as a sampling device to collect both hoverflies and bees. Other than altitude, the effect of landscape complexity and plant diversity were considered as potential predictors of hoverfly and bee richness and abundance along the two gradients. A total of 68 species of hoverflies and 67 of bees were collected during one sampling year, confirming the efficacy of pan traps as a sampling device to study these taxa. Altitude was the main variable affecting both hoverfly and bee distribution. The two taxa show different distribution patterns: hoverflies have a unimodal distribution (richness and abundance) with peak at middle altitude (1500 m), while bees have a monotonic decline (richness and abundance) with increasing altitude. Both hoverfly and bee populations change with the increasing altitude, but the change in hoverflies is more pronounced than in bees. Species turnover dominates the β-diversity both for hoverflies and bees; therefore, the hoverfly and bee communities at higher altitudes are not subsamples of species at lower altitude but are characterized by different species. This poses important conservation consequences. Some rare species, typical of an alpine habitat were recorded; the present research represents important baseline data to plan a monitoring scheme aimed at evaluating the effect of climate change on pollinators in these fragile habitats.
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Apple production in the UK is worth over £100 million per annum and this production is heavily dependent on insect pollination. Despite its importance, it is not clear which insect pollinators carry out the majority of this pollination. Furthermore, it is unknown whether current UK apple production, in terms of both yield and quality, suffers pollination deficits and whether production value could be increased through effective management of pollination services. The present study set out to address some of these unknowns and showed that solitary bee activity is high in orchards and that they could be making a valuable contribution to pollination. Furthermore, fruit set and apple seed number were found to be suffering potential pollination deficits although these were not reflected in apple quality. Deficits could be addressed through orchard management practices to improve the abundance and diversity of wild pollinators. Such practices include provision of additional floral resources and nesting habitats as well as preservation of semi-natural areas. The cost effectiveness of such strategies would need to be understood taking into account the potential gains to the apple industry.
Technical Report
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Description Fit linear and generalized linear mixed-effects models. The models and their components are represented using S4 classes and methods. The core computational algorithms are implemented using the 'Eigen' C++ library for numerical linear algebra and 'RcppEigen' ``glue''.
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There is compelling evidence that more diverse ecosystems deliver greater benefits to people, and these ecosystem services have become a key argument for biodiversity conservation. However, it is unclear how much biodiversity is needed to deliver ecosystem services in a cost-effective way. Here we show that, while the contribution of wild bees to crop production is significant, service delivery is restricted to a limited subset of all known bee species. Across crops, years and biogeographical regions, crop-visiting wild bee communities are dominated by a small number of common species, and threatened species are rarely observed on crops. Dominant crop pollinators persist under agricultural expansion and many are easily enhanced by simple conservation measures, suggesting that cost-effective management strategies to promote crop pollination should target a different set of species than management strategies to promote threatened bees. Conserving the biological diversity of bees therefore requires more than just ecosystem-service-based arguments.
A guide to using S environments to perform statistical analyses providing both an introduction to the use of S and a course in modern statistical methods. The emphasis is on presenting practical problems and full analyses of real data sets.
In recent years, an intense debate has been generated about the environmental risks posed by neonicotinoids, a group of widely-used, neurotoxic insecticides. When these systemic compounds are applied to seeds, low concentrations are subsequently found in the nectar and pollen of the crop, which are then collected and consumed by bees. Here we demonstrate that current focus on exposure to pesticides via the crop overlooks an important factor - throughout spring and summer, mixtures of neonicotinoids are also found in the pollen and nectar of wildflowers growing in arable field margins, at concentrations that are sometimes even higher than those found in the crop. Indeed the large majority (97%) of neonicotinoids brought back in pollen to honey bee hives in arable landscapes was from wildflowers, not crops. Both previous and ongoing field studies have been based on the premise that exposure to neonicotinoids would only occur during the blooming period of flowering crops and that it may be diluted by bees also foraging on untreated wildflowers. Here, we show that exposure is likely to be higher and more prolonged than currently recognized due to widespread contamination of wild plants growing near treated crops.