The floral and the geographical origins of honey can be determined

by microscopic examination of the pollen present, however

Neonicotinoids are registered globally in more than 120 countries, and they are among the most
effective insecticides available for control of sucking insect pests such as aphids, white flies,
leaf- and plant-hoppers, thrips, some micro-lepidoptera, and a number of coleopteran (beetle)
pests. They have broad-spectrum activity, with contact, stomach and systemic activity. Their
physicochemical properties mean that they are very versatile in terms of application
methodology, being used for foliar, seed treatment, soil drench and stem applications in a wide
range of crops.

The neonicotinoids are the most significant chemical class of insecticides introduced to the
global market since the synthetic pyrethroids.

Imidacloprid was the first neonicotinoid insecticide to come into commercial use, in 1991. It is
currently the most widely used insecticide worldwide. It is applied against pests in/on soil, seed,
timber and animals, and is in products for foliar application to turf and food crops including
cereals, cotton, legumes, potatoes, pome fruits and vegetables. It is a systemic insecticide with
particular efficacy against sucking insects and has quite a long residual activity. Because of its
water solubility it can be added to the water used to irrigate plants.

In 1990 (before the launch of imidacloprid), the insecticide market was dominated by
organophosphorus insecticides (‘organophosphates’ or OPs) (ca. 43%), pyrethroids (ca. 18%)
and carbamates (ca. 16%). By 2008, neonicotinoids had gained around 24% share of the total
insecticide market, mainly at the expense of the OPs and carbamates (which fell to around 13.6%
and 10.8% of the insecticide market, respectively, at this time). In the past several years the use
of neonicotinoid insecticides has grown further; in 2009 imidacloprid was estimated to account
for ca. 41.5% of the global neonicotinoid market, making it the largest selling insecticide in the
world. Thiamethoxam was second in terms of sales, followed closely by clothianidin. Together,
the N-nitroguanidine class of neonicotinoids account for around 85% of the neonicotinoid
insecticide market. Nevertheless, sales of other neonicotinoids have grown as well (Jenschke et
al, 2011).

The application rates for neonicotinoid insecticides are commonly much lower than older classes
of insecticides (eg. Jeschke et al, 2010).

When compared with OPs and carbamates, neonicotinoids pose lower risks to humans and other
mammals.
As a result of their mechanism of action, there is no cross-resistance to other insecticide classes,
including the so-called organochlorines (now superseded), OPs, carbamates and pyrethroids.
Their introduction has increased the insecticide armentarium available to farmers and
horticulturalists, thus helping to prevent the build-up of resistance of insect pests to OPs and
pyrethroids. Nevertheless, pests can evolve resistance to neonicotinoids; the first reported
example was the silverleaf whitefly, Bemisia tabaci (Thany, 2010).

One problem with systemic insecticides is that the selection pressure placed on pests from the
ongoing presence of the insecticide, coupled with their site-specific, mode of insecticidal action,
may result in the development of resistant insect genotypes. Conversely, the benefits of using
systemic insecticides like the neonicotinoid seed-treatment insecticides include (1) plants are
continuously protected throughout most of the growing season without the need for repeat spray
applications of insecticides; (2) these insecticides are not susceptible to UV light degradation or
‘wash off’ during watering; (3) there is no surface residue on the crop and hence a reduced risk
to agricultural workers performing in-crop activities.

A comprehensive review of the value of neonicotinoid seed treatments has been published by the
Humboldt Forum for Food and Agriculture e.V. (Noleppa & Hahn, 2013). It investigated the
socioeconomic and environmental contribution made by this technology to the European Union
across major crops and key countries, and the significant economic and social impact should the
technology no longer be available because of bans or suspensions.

Neonicotinoids are registered globally in more than 120 countries, and they are among the most
effective insecticides available for control of sucking insect pests such as aphids, white flies,
leaf- and plant-hoppers, thrips, some micro-lepidoptera, and a number of coleopteran (beetle)
pests. They have broad-spectrum activity, with contact, stomach and systemic activity. Their
physicochemical properties mean that they are very versatile in terms of application
methodology, being used for foliar, seed treatment, soil drench and stem applications in a wide
range of crops.

The neonicotinoids are the most significant chemical class of insecticides introduced to the
global market since the synthetic pyrethroids.

Imidacloprid was the first neonicotinoid insecticide to come into commercial use, in 1991. It is
currently the most widely used insecticide worldwide. It is applied against pests in/on soil, seed,
timber and animals, and is in products for foliar application to turf and food crops including
cereals, cotton, legumes, potatoes, pome fruits and vegetables. It is a systemic insecticide with
particular efficacy against sucking insects and has quite a long residual activity. Because of its
water solubility it can be added to the water used to irrigate plants.

In 1990 (before the launch of imidacloprid), the insecticide market was dominated by
organophosphorus insecticides (‘organophosphates’ or OPs) (ca. 43%), pyrethroids (ca. 18%)
and carbamates (ca. 16%). By 2008, neonicotinoids had gained around 24% share of the total
insecticide market, mainly at the expense of the OPs and carbamates (which fell to around 13.6%
and 10.8% of the insecticide market, respectively, at this time). In the past several years the use
of neonicotinoid insecticides has grown further; in 2009 imidacloprid was estimated to account
for ca. 41.5% of the global neonicotinoid market, making it the largest selling insecticide in the
world. Thiamethoxam was second in terms of sales, followed closely by clothianidin. Together,
the N-nitroguanidine class of neonicotinoids account for around 85% of the neonicotinoid
insecticide market. Nevertheless, sales of other neonicotinoids have grown as well (Jenschke et
al, 2011).

The application rates for neonicotinoid insecticides are commonly much lower than older classes
of insecticides (eg. Jeschke et al, 2010).

When compared with OPs and carbamates, neonicotinoids pose lower risks to humans and other
mammals.

As a result of their mechanism of action, there is no cross-resistance to other insecticide classes,

including the so-called organochlorines (now superseded), OPs, carbamates and pyrethroids.
Their introduction has increased the insecticide armentarium available to farmers and
horticulturalists, thus helping to prevent the build-up of resistance of insect pests to OPs and
pyrethroids. Nevertheless, pests can evolve resistance to neonicotinoids; the first reported
example was the silverleaf whitefly, Bemisia tabaci (Thany, 2010).

One problem with systemic insecticides is that the selection pressure placed on pests from the
ongoing presence of the insecticide, coupled with their site-specific, mode of insecticidal action,
may result in the development of resistant insect genotypes. Conversely, the benefits of using
systemic insecticides like the neonicotinoid seed-treatment insecticides include (1) plants are
continuously protected throughout most of the growing season without the need for repeat spray
applications of insecticides; (2) these insecticides are not susceptible to UV light degradation or
‘wash off’ during watering; (3) there is no surface residue on the crop and hence a reduced risk
to agricultural workers performing in-crop activities.

A comprehensive review of the value of neonicotinoid seed treatments has been published by the
Humboldt Forum for Food and Agriculture e.V. (Noleppa & Hahn, 2013). It investigated the
socioeconomic and environmental contribution made by this technology to the European Union
across major crops and key countries, and the significant economic and social impact should the
technology no longer be available because of bans or suspensions.

OVERVIEW REPORT NEONICOTINOIDS AND THE HEALTH OF HONEY BEES IN

AUSTRALIA. FEBRUARY 2014. Australian Pesticides and Veterinary Medicines Authority
2013

Pollution and other threats

Air pollution hampers the symbiotic relationships between pollinators and flowers.
Although daytime insects depend primarily on vision to find flowers, pollutants affect
the chemicals that flowers produce to attract insects, which destroys vital scent trails.
Scents that could travel over 800m in the 1800s now reach less than some 200m
from the plant, which complicates pollinators’ ability to locate food sources31. McFrederick Q.
S., Kathilankal J. C. et al. 2008. “Air pollution modifies floral scent trails”. Atmospheric
Environment 42, 2336–2348.
Chemical drifts from spraying
Chemicals can poison pollinators or impair their reproduction, eliminate nectar
sources and destroy larval host plants for moths and butterflies and deplete bees’
nesting materials32. It is plausible that plant losses from chronic herbicide use may
be driving losses of pollinator species33. Additionally, various broad-spectrum
insecticides are not only applied on agricultural fields but also in residential
gardens, recreational areas, forests as well as mosquito-ridden marshes and
swamps. These chemicals can be equally toxic to beneficial insects as to the target
species34. Chronic or sub-lethal exposure to agricultural or beekeeper-applied
pesticides can weaken the honey bee’s immune system, and hamper bees’ ability to
fight infection. The indirect effects of pesticides on pollinator populations, particularly the
destruction of valuable plants and habitats after herbicide spraying, must also be
considered. The chemical destruction of habitats can have long-term consequences
particularly on the distribution of pollinators in agro-environments.

32.Nabhan G.P. and Buchmann S.L, 1997. “Pollination services: biodiversity’s direct link to world food stability”, in G. Daly, ed.
Ecosystem Services, Island Press, Washington, D.C.
33 Cane J. H. and V. J. Tepedino. 2001. “Causes and extent of declines among native North American invertebrate pollinators:

detection, evidence, and consequences”. Conservation Ecology 5(1): 1 at www.consecol.

org/vol5/iss1/art1/
34 Johansen C. A., and Mayer D. F. 1990. “Pollinator protection: a bee and pesticide handbook”. Wicwas Press, Cheshire,

Connecticut, USA.
35 Cox C, 2001. “Imidacloprid, Insecticide factsheet”. Journal of Pesticide Reform, Vol. 21, N°1 at www.pesticide.org/imidacloprid.pdf

36 Bonmatin, J.M., Marchand P.A, Charvet R, Colin M.E, 1994. “Fate of systemic insecticides in fields (Imidacloprid and Fipronil) and

risks for pollinators, in First European Conference of Apidology”. Udine 19-23 September
2004.
37 Colin M. E., Bonmatin J. M. et al. 2004. “Quantitative analysis of the foraging activity of honey bees: relevance to the sub-lethal

effects induced by systemic insecticides”. Arch. Environm. Contam. Toxicol. 47, 387-395.
38 Bonmatin J. M., Marchand P.A. et al. 2005. “Quantification of imidacloprid uptake in maize crops”. Journal of agricultural and food

chemistry , vol. 53, no13, pp. 5336-5341.

The toxicity and biochemical changes in honey bees (Apis mellifera ) treated with four
insecticides—acetamiprid, dinotefuran, pymetrozine, and pyridalyl—were evaluated under controlled laboratory
conditions. Foraging bees were exposed to different dosages of tested insecticides by oral feeding at different
dosages recommended by the manufacturers for agricultural crops in Egypt (0.01-, 0.02-, 0.04-, 0.1-, and onefold).
Moreover, the acute toxicity of these insecticides was evaluated by topical application on the thorax of foragers to
calculate the LD50 values. The specific activities of acetylcholinesterase (AChE), carboxylesterase, glutathione Stransferase
(GST), and polyphenol oxidase (PPO) were measured in different tissues of surviving foragers after 24 h
of treatment to explore the possible mode of action of insecticides and honey bees' strategies for detoxification and
tolerance. The results indicated that regardless of how the bees were exposed to insecticides, dinotefuran was
extremely toxic to adult A. mellifera (topical LD50 = 0.0006 μg/bee and oral feeding LC50 = 1.29mg/L). Pyridalyl
showed moderate toxicity compared to dinotefuran at the recommended application rate; however, acetamiprid and
pymetrozine were relatively less toxic to bees (<25%mortality at the recommended application rates). Data showed
that tested insecticides varied in their influence on AChE, carboxylesterase,GST, and PPOactivities that were highly
correlated to their toxicity against A. mellifera . The biochemical analysis of carboxylesterase and GST showed that
these enzymes detoxified the low doses of acetamiprid, pymetrozine, and pyridalyl, but not dinotefuran. Overall, our
results are valuable not only in evaluating the toxicity of common insecticides onto honey bees, but also in
highlighting the validity of enzymes activities as proper indicators for exposure to agrochemicals.

Analysis by Liquid Chromatography-Mass Spectrometry

Although the extraction steps are efficient and selective, it is necessary to perform a
separation by high performance liquid chromatography. Imidacloprid is thermo-labile,
thus HPLC allows direct injection of the extract, whereas gas chromatography
(GC) requires derivatization. The detection by mass spectrometry coupled to LC allows
clear identification of the parent ion at m/z = 256, a very high specificity using
two or more ions products (m/z = 209 and m/z = 175), and a very low limit of detection
as detailed in the next section.
J. M. Bonmatin · I. Moineau · R. Charvet · M. E. Colin · C. Fleche · E. R. Bengsch

Antipyrine was systematically used as an external standard between 10

injections of unknown samples. Unknown samples were also injected
between series of stan-dards and quality control samples. The retention
time was constant with a deviation never exceeding 2.5%. The retention
time was typically 2.4 min for the plants and 2.8 min for the pollen and soils
samples depending on the chosen chromatographic flow. Calibration graphs
were plotted for six standard solutions between 0.5 and 20 µg kg–1 of
imidacloprid added in extracted materials (blanks). For each calibration
point, three injections into the liquid chromatograph were performed.
Linearity was found in the range between 0.5 and 20 µg kg–1. The
correlation coefficients were bet-ter than 0.99. Fig. 44.2. Extraction schemes
applied to soils, plants and pollens prior the separation and detection processes. Schemes
were suited to obtain high recovery levels of imidacloprid at the level of the limit of
quantification (1 µg kg–1)

N
Extraction schemes applied to soils, plants and pollens prior the separation and detection
processes. Schemes were suited to obtain high recovery levels of imidacloprid at the level of the limit
of quantification (1 µg kg–1)
Antipyrine was systematically used as an external standard between 10 injections of unknown samples.
Unknown samples were also injected between series of standards and quality control samples. The retention
time was constant with a deviation never exceeding 2.5%. The retention time was typically 2.4 min for the
plants and 2.8 min for the pollen and soils samples depending on the chosen chromatographic flow. Calibration
graphs were plotted for six standard solutions between 0.5 and
20 µg kg–1 of imidacloprid added in extracted materials (blanks). For each calibration
point, three injections into the liquid chromatograph were performed. Linearity was
found in the rang