Journal of Phytopathology and Pest Management 7(1): 31-42, 2020
pISSN:2356-8577 eISSN: 2356-6507
Journal homepage: http://ppmj.net/
∗
Corresponding author:
Gamal A. Abdu-Allah,
E-mail: gamalan@aun.edu.eg
31
Copyright © 2020
Comparison between the toxicity of nano and bulk
formulations of imidacloprid against wheat aphid,
bird cherry-oat aphid,
Rhopalosiphum padi
L
Hassan M. Abd Elwahab
1
, Youssef A. Darwish
1
, Hosam El-Din A. Ezz El-Din
1
, Ali A. Othman
2
, Gamal A. Abdu-Allah
1*
1
Plant Protection Department, Faculty of Agriculture, Assiut University, 71526 Assiut, Egypt
2
Physics Department, Faculty of Science, Assiut University, 71526 Assiut, Egypt
Abstract
Keywords: nano- insecticides, toxicity, seed treatments, grain yield.
32
1. Introduction
Wheat,
Triticum aestivum
L. (Family:
Poaceae) is one of the most important
food crops for human and animal feed.
The Egyptian cultivated area of the wheat
crop is around 3,258969 million feddans,
producing about 9 million tons in Egypt
(FAO, 2016). Wheat crop is
attacked by
many insect pests that cause great
damages
and
reductions in the yield
.
Five
cereal
aphid species (
Rhopalosiphum
padi, Schizaphis graminum,
Rhopalosiphum maidis, Sitobin avenae,
Metopolophium dirhodum
are the most
serious key insect pests attacking wheat
in Upper Egypt
.
(Abd-Ella, 2016; Abdel-
Rahman, 1997). In addition to, the direct
damages of aphids by producing
honeydew that encouraging the growth of
softy moulds, which considerably
interface with
the
host photosynthesis
process by inserting their stylet on plant
cells and tissues, triggering reactions and
cell death by the toxins and enzymes in
their saliva, and by sucking plant sap.
Rhopalosiphum padi
causes indirect
damages by transmission of
Barley/Cereal Yellow Dwarf Virus
(B/CYDVs) the causal agents of Yellow
Dwarf Disease (YDD) (Parizoto et al
.
,
2013). Insecticides are a widely
traditional method used for controlling
the cereal aphids. Foliar application of
broad-spectrum insecticides may reduce
natural enemies(Galvan
et al.
,
2005),contributing to pest resurgence.
Seed treatment, another application
method, with neonicotinoids has
advantages especially the long protection
time when compared to certain
application of pesticides, particularly
when used to control insect pests that
feed on seeds or seedlings early in the
season. Moreover, the neonicotinoids
have a combination of high toxicity to
insects and low toxicity to mammals that
make them safer than many of the older
insecticides. Seed application allows the
amount applied per area to be greatly
reduced compared to soil or foliar
applications ( Nuyttens et al., 2013;
Elbert et al., 2008). Imidacloprid is
belong to neonicotinoid group of
chemicals which act as systemic
insecticide against sucking pests. The
neonicotinoids have quick knockdown
effect on target pest by interfering with
transmission of impulse in the nerve
system. The quick and excellent systemic
and translaminar activity of these
insecticides make them able to control
sucking pests such as aphids, whiteflies
and other virus transmission insects.
Neonicotinoid application reduces
infection rate and spread of many crop
viruses (Bethke
et al.,
2001; Elzen, 2001;
Elbert
et al.
, 1998; Westwood
et al.,
1998; Dewar, 1992; Knaust and
Poehling, 1992). Due to the side effects
of conventional insecticide, the
development of the insecticide
application and insecticide formulation
are valuable trend to alleviate these bad
effects. Nanotechnology can introduce
promised nano-insecticides that have so
different physicochemical prosperities.
Few studies are available regarding the
toxicity of these nano-insecticides on
insects. The objective of the present
study was taken to compare between the
toxicity of nano and bulk formulations of
imidacloprid against bird cherry-oat
aphid,
R. padi
under field and laboratory
conditions. Moreover, the effect of the
seed treatments of the tested insecticides
on grain yield crop.
2. Materials and methods
2.1 Insecticides nano preparation
Two insecticide imidacloprid
formulations (Best 25% WP, EL-Helb
Pesticides & Chemicals, Egypt and
33
imidacloprid Gaucho 70% WS, Bayer
Crop Science, Egypt) were supplied from
the Central Pesticide Laboratory, Egypt.
The insecticide powders were milled in a
FRITSCH PULVERISETTE
2
ball mill
for 1 hour to obtain the micro-size
formulations. X-ray diffraction (XRD) of
the prepared nano formulations were
recorded using PW 1700 X-ray
diffraction with Cu
α
K radiation (λ =
1.5406 angstrom) in diffraction angle (2
θ) range from 20
o
to 60
o
with step
0.06
o
/min. Figures (1, 2) represents X-ray
diffraction pattern of the treated powder.
The line broadening of the XRD peaks
indicates that the prepared material
consists of particles in the nano-size
range. The pesticide particle size was
calculated from–Scherrer’ equation:

(0.89 is Scherrer’s constant, λ is the
wavelength of X-rays,( θ )is the Bragg
diffraction angle, and( β) is the full width
at half-maximum (FWHM) of the
diffraction peak .The average particle
size of was found to be 21.8 nm for best
and 18.1nm for Gaucho.
Figure 1: X-rays diffraction of treated insecticide
imidacloprid 25% WP.
Figure 2: X-rays diffraction of treated insecticide
imidacloprid 70% WS.
2.2 Insecticide laboratory bioassay
tests
The relative effectiveness of nano and
bulk formulations of imidacloprid
neonicotinoid insecticide was tested
under laboratory conditions.
2.3 Test insect
The field population of the oat aphid,
Rhopalosiphum padi
was collected from
infested field wheat batches under Assiut
prevailing conditions. The apterous
aphids used in this study were of
approximately the same size and weight
for all the toxicity tests. Fresh infested
wheat samples were collected by cutting
plants and transferred alive to laboratory
on their host plant leaves in plastic bags
immediately before assays where mean
temperature reached about 26 ± 2
º
C. The
leaf dip-bioassay technique (O’brien et
al., 1992) was followed to evaluate the
toxicity of insecticides. Five to six
concentrations of each insecticide plus
commercial surfactant (frotop®) were
34
prepared. About 10 to 15 adults,
approximately of same size, were dipped
for five seconds per concentration. The
treated aphids could dry at room
temperature for about half an hour.
Control batches of aphids were similarly
dipped in a solution of water plus
(frotop
®
). After the treated batches of
aphids had dried they were individually
transferred to petri dishes (9 cm
diameter) and held for 26 ± 2Cº, 60 ± 5
RH% and photoperiod 12:12 (L:D).
Aphid mortality was recorded after 12,
24 and 48 h for treatments with the aid of
a binocular microscope. An aphid was
considered dead if it was in cap-able of
coordinated forward movement and
changing of color body to black. The
toxicity experiment of each compound
was repeated twice, and the results were
corrected by Abbott
’
s formula (Abbott,
1925). Median lethal concentration
(LC
50
) and slope evaluation of the tested
insecticides were determined by Probit
regression analysis program.
2.4 Preparation of imidacloprid
formulations as seed treatment
Seeds of the wheat variety Sids (12) were
put in a plastic container. Each
insecticide formulation was mixed with
the seeds at rate of 9.8 g a.i kg
-1
seeds, it
is equal the recommended rate by
imidacloprid (Best
®
25%WP), and 3.5 g
a.i. kg
-1
seeds (it is equal the
recommended rate by imidacloprid
(Gaucho® 7 g kg-1 seeds) as
manufacture label. Gaucho®, the only
neonicotinoid insecticide recommended
as seed treatments in certain crops. The
control seeds were treated with water.
Carefully shucked in then the seeds were
gently participated on the desk left in air
to dry in shade.
2.5 Field experiment and data
collection
A field study was conducted to compare
the efficacy and persistence of the nano-
imidacloprid formulations with
conventional (Bulk) formulations against
cereal aphids. On 10
th
December, 2018,
the treated seeds of wheat variety Sids
(12) were cultivated in plots at the
Experimental Farm of Faculty of
Agriculture, Assiut university, Egypt.
The area of each plot was 1/400 feddan
(0.42 hectare). A randomized complete
block design (RCBD) with 3 replicates to
every treatment and 3 untreated
replicates as control was used. After 22
days from planting, the aphid appeared
on the wheat plants, the wheat aphid,
R.
padi
began to occupy on the upper and
lower leaves of wheat plants. The
number of wheat aphid on 5 randomly
selected wheat tillers from five locations
per plot was recorded. The wheat aphid
populations were weekly recorded until
the maturity of the wheat plant. The
aphid species identification was done
using binocular in the Insect Laboratory,
Plant Protection, Faculty of Agriculture,
Assiut University, Egypt. The percentage
(%) reduction of insect infestation was
calculated according to a modified
version of Abbott
’
s formula (Abbott,
1925):
Reduction (%)  


 
Where; T = treatment, C = control, N=
the number of aphids.
Grain yield: Average yield of grain
wheat (kg /feddan) (feddan = 0.420
35
hectares = 1.037 acres) was assessed
using the harvest from the treated plots
and the control of wheat crops.

  


3. Results and Discussion
The toxicity of two bulk-imidacloprid
formulations (Best
®
and Gaucho
®
) and
their prepared nano-formulations were
investigated against the apterous adults of
R. Padi
using the leaf-dip bioassay under
laboratory conditions. Based on the LC
50
values and the overlapping at 95%
confidence limits represented in Tables
(1 & 2), data indicated that most of bulk
and nano imidacloprid formulations (WS
Gaucho
®
) showed significant higher
toxicity against wheat aphids than the
analogous formulations of imidacloprid
(WP Best
®
) after 12, 24 hours of the
treatment, where, the LC
50s
values of
Gaucho
®
bulk and Best
®
bulk
formulations were 1406.78, 4018.83;
608.92, 1584.23 µg a.i./ml after 12, 24
hours, respectively. Whereas, the
opposite is true after 48 hours post
treatment, the Best
®
formulations have
the higher toxicity effect than the
Gaucho
®
formulations. In general, the
toxicity of all imidacloprid formulations
is dependently increased with the
increased time after treatment.
Table 1: Comparison the toxicity of bulk and nano-Imidacloprid neonicotinoid insecticide (BEST
®)
against
the adult apterous of bird cherry-oat aphid, Rhopalosiphum padi after 12, 24,48 h post treatment using the
leaf-dip bioassay.
Time
(Hour)
Bulk-Imidacloprid 25%WP(BEST
®
)
Nano-Imidacloprid 25%WP(BEST
®
)
Potency Ratio
Slope ±SE
χ2
P
LC
50
(µg a.i./ml)
(Confidence Limits 95%)
Slope±SE
χ
2
P
12
1.09
±0.10
9.64
0.140
1585.70
(1289.94-2028.84)
1.45
±0.11
6.19
0.402
2.53
24
1.03
±0.08
14.55
0.024
253.43
(126.80-501.66)
0.54
±0.06
10.72
0.097
6.25
48
0.68
±0.07
3.08
0.799
7.470
(1.91-16.72)
0.53
±0.07
1.97
0.922
11
Potency ratios:LC
50
of Bulk-Imidacloprid ∕LC
50
of Nano-Imidacloprid; Chi-square = χ
2
value.
Table 2: Comparison the toxicity of bulk and nano-Imidacloprid neonicotinoid insecticide (GAUCHO
®)
against the adult apterous of bird cherry-oat aphid, Rhopalosiphum padi after 12, 24, 48 h post treatment
using the leaf-dip bioassay.
Time
(Hour)
Bulk-Imidacloprid 70%WS (GAUCHO
®
)
Nano-Imidacloprid 70%WS (GAUCHO
®
)
Potency Ratio
Slope ±SE
χ2
P
LC
50
(µg a.i./ml)
(Confidence Limits 95%)
Slope±SE
χ
2
P
12
0.61
±0.07
6.07
0.415
271.45
(200.45-392.59)
0.76
±0.07
4.74
0.577
5.18
24
0.39
±0.06
2.49
0.869
129.57
(91.44-190.88)
0.59
±0.06
7.10
0.311
5
48
0.38
±0.06
3.47
0.748
18.44
(10.05-28.51)
0.56
±0.07
4.93
0.552
7.17
Potency ratios: LC
50
of Bulk- Imidacloprid ∕LC
50
of Nano- Imidacloprid, Chi-square = χ
2
value.
The toxicity data (Tables 1 & 2) showed
that the nano formulation of imidacloprid
(Best
®
) had significantly higher potency
effect than its bulk formulation by 2.53,
6.25 and 11 folds respectively, after 12 h,
24 h, and 48 h post treatment. Also, the
36
nano formulation of imidacloprid
(Gaucho
®
) had significant higher potency
than its bulk formulation by 5.18, 5 and
7.17 folds. Iimidacloprid and
thiamethoxam had a better efficiency
against wheat aphids than acetamiprid
and dinotefuran and caused an increase in
wheat production (Abd-Ella, 2016). In
the recent years, nano particles have
received much attention for controlling as
agriculture pathogens. The quick and
increased the toxicity of imidacloprid
after time post treatment against wheat
aphids could be explained imidacloprid is
one of the neonicotinoid insecticide
group. Neonicotinoids act as neurotoxic
effect of on nicotinic acetylcholine
receptors (nAchR) (Tomizawa and
Casida, 2005)
.
In the present results, the
variation in potency effects of the Best
®
and Gaucho
®
formulations depends on
several factors such as the insecticide
formulations, the particle size,
penetration of the plant, and the
prevailing environmental conditions
during experimental seasons. All these
factors can play a role on the pesticide
disappearance among plants and
influencing the efficiency of the tested
insecticide. In the other hand nano-
formulation increases the efficiency of
insecticides and also reduces the dose
level required to pest control work. In
addition, pesticide permeability during
the layers outside the surface of the plant
depends on the amount of pesticides used
in the plant tissues, and on the rate of
demolition, which also varies greatly
depending on the chemical composition
of the compound tested under the field
conditions (Saad et al., 2019; Farha et al.,
2016). Little research that had been
carried out to investigate the toxicity
effect of nanoparticles on insects.(Guan
et al.,
2008) showed that toxicity of the
bulk formulation of imidacloprid
increased when coated with nano
particles. Our results are supported by
Assemi
et al. (2014) who found that the
LC
50
values for nano imidacloprid, were
37.919, 69.623 μl/ ml, respectively after
72 hours. Their results showed that nano-
pesticides are more effective and toxic to
insect pests, compared to conventional
insecticides. On the other hand, (Rouhani
et al., 2012) were evaluated the
insecticidal activity of silver
nanoparticles against the
Aphis nerii
.
They found that Ag nanoparticles can be
used as a valuable tool in pest
management programs of
A
.
nerii
. Our
laboratory results are promised; these
results need more investigation under the
field conditions. It is of interest to point
herein that the nano-neonicotioid
insecticides showed great may be
reducing effect on
Liriomyza trifolii
(Burgess) tested than their bulk
formulation tested insecticides(Saad et
al., 2019). The manufacture
recommended dose of imidacloprid
formulation (1X) and the half dose (0.5
X) of bulk and nano of Best
®
and
Gaucho
®
formulations evaluated as a
seed treatment for reducing and delaying
the
R. padi
infestation of wheat plants in
the field. The aphid’s infestation on
wheat crop started after 60 days, of
sowing. The aphid population was
observed during vegetative stage and
continued up to last week of February.
The the bulk and nano formulations of
imidacloprid in all treatment showed
significant lowest aphid population as
compared to untreated (Control) after 60,
74 and 102 days of sowing. In significant
difference has occurred in reduction %
between the insecticide treatments. Also,
37
the half dose of nano and bulk
imidacloprid formulations gave positive
results in reducing % (reaching 95%) of
aphids on wheat crops (Figures 3 & 4).
Figure 3: The mean of aphids/tiller at 60(A), 74(B), and
102(C) days of sowing. The error bars indicate the standard
deviation of means. The figures with the same letter
showed insignificant differences at 5% probability level,
LSD test.
Figure 4: The mean reduction percentage of aphids at
60(A), 74(B), and 102(C) days of sowing. The error bars
indicate the standard deviation of means. The figures with
the same letter showed insignificant different from each
other at 5% probability level, LSD test.
Burd et al. (1996) reported acute
toxicity to aphids fed on plants grown
from seeds treated with imidacloprid,
protected plants for 45 days after
sowing. While, imidacloprid in
combination with tebuconazole can
control aphids for 8 weeks when applied
as a seed treatment by Ahmed et al.
(2001). Our results are also in
agreement with the results Suhail
et al.
(2013) who found that aphid population
was lowest on plants of wheat which
were grown with seed treated with
Hombre and Actara. Our results are also
in agreement with those of Royer et al.
(2005) who found that the seed
treatment with imidacloprid and other
insecticides decreased the population of
sucking insect pests such as cereal
aphids and leaf hoppers (Khan et al.,
2012; Zeb et al., 2011; Aheer et al.,
2006; Aslam et al., 2004). Several
investigators demonstrated that the
aphid infestation on wheat crop started
in the first week January 60 days after
sowing. A gradual increase in aphid
population was observed during
vegetative stage and continued up to
last week of February. On the other
hand, the higher yield was observed in
plots treated with imdacloprid +
tebuconazole followed by dinotefuran,
38
matrine, and pymetrozine respectively
by (Ali et al.,
2018). The results of grain
yield are influenced by different
insecticide treatments ( Figure 5 ) where
the grain yield production per feddan
treated with Bulk imidacloprid Best
®
(1X), Nano imidacloprid Best
®
(1X),
Bulk imidacloprid Best
®
(0.5X), Nano
imidacloprid Best
®
(0.5X), and
imidacloprid Gaucho
®
(1X) was 3204,
3236, 3192, 2932, and 2932 kg/feddan,
respectively compared with the control
(2652 kg/feddan). Subsequently, yield
increasing as compared to untreated
(Control) percentage was 20.81, 22.02,
20.51, 21.11, and 10.55 %, respectively.
Figure 5: Estimated average seed yield of grain wheat (3A) and
increase percentage of yield(3B) due to imidacloprid
neonicotinoid insecticide formulations seed treatments compared
to the control. The error bars indicate the standard deviation of
means. The figures with the same letter showed insignificant
differences at 5% probability level, LSD test.
In a similar study, the imidacloprid seed
treatment enhanced the grain yield when
used against Russian wheat aphids
Moreover, in our results the half dose of
nano and bulk imidacloprid formulations
showed positive results in grain yield of
A
B
39
wheat crops (Tolmay et al., 1997). The
results of the present study are in
accordance with the findings of (Royer
et
al., 2005) who achieved 100% morality
of aphids through some botanical
insecticides, this may be due to
difference in crop and environmental
factors as they tested on wheat plants. In
addition imidacloprid was most effective
with maximum mortality 97% and
pymetrzine 87%, respectively after 14
days of application (Shafiq et al., 2015).
VoSTřel (2010) found 100% mortality in
resistant population of wheat aphids
when treated with imidacloprid. Another
study on effectiveness of imidacloprid
and other insecticides indicated that
imidacloprid-treated oat or wheat plants
reduced adult longevity and fecundity of
three cereal aphid species as compared to
non-treated plants (Gray et al., 1996).
Imidacloprid 200 SL at ml/ha treatment
was most effective against wheat aphid
Sitobion avenae
,
Rhopalosiphum maidis
(Joshi and Sharma, 2009).
Our finding
are supported by other publications
especially on faba bean seed treatments
(Abdu-Allah and Hashem, 2017) who
found that imidacloprid and
thiamethoxam and acetamiprid as
cowpea seed treatment significantly
increased the cowpea height growth
compared to untreated control. However,
the longevity, fertility and population
increase of
S. avenae
feeding on wheat
plants treated with sublethal
concentrations (LC
10
) of imidacloprid
was not greatly affected, and the
longevity, fertility and population
increase of
S. avenae
reduced
significantly at the median lethal
concentration (LC
50
) of the four
neonicotino (Miao et al., 2014). Lower
fecundity has also been observed in
Aphis gossypii
after treatment with
imidacloprid (Gerami et al., 2005). Also,
imidacloprid was effective in reducing
the population of aphids on brinjal
(auberggine) and in increasing the
seedlings total leaf chlorophyll over
those of untreated plants (Jarande and
Dethe, 1994). The present results are in
same line with those reported by (Abdu-
Allah, 2012), who studied the toxicity of
imidacloprid on cowpea aphid,
A.
craccivora
and
B. brassicae,
after 48 h
using leaf-dip bioassay technique. He
stated that imidacloprid was the least
toxic compound against
A. craccivora
with LC
50
of
2.04 mg/L and against
B.
brassicae
with LC
50
of 2.14 mg/L.
4. Conclusions
Our results concluded that the nano
formulation of imidacloprid was more
potency effective and toxic to wheat
aphids,
R.padi
, compared to
conventional insecticides. These studies
clearly showed the need to research and
development under the field conditions.
Also the half dose of nano and bulk
imidacloprid formulations give positive
results in reducing percentage of aphid
and its increasing yield percentage by
about 22.02%. Seed treatment provides
excellent control at early stage of
infestation by aphid, good plant vigor
and thus greater yield and less residues
with long plant protection for 42 days
after sowing. Conversely, their
management in the later stage of
infestation (foliar treatment).
40
Acknowledgements
Thanks a lot, to Dr. Alhosein Hamada
Abd-El-Azeem Hassan, Associate
Professor, Department of Agronomy for
the assistance of the statistical analysis of
the data.
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