Physiological and Molecular Plant Pathology xxx (2017) 1e11

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Physiological and Molecular Plant Pathology

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Suaeda maritima-based herbal coils and green nanoparticles as

potential biopesticides against the dengue vector Aedes aegypti and
the tobacco cutworm Spodoptera litura
Udaiyan Suresh a, Kadarkarai Murugan a, b, Chellasamy Panneerselvam c,
Rajapandian Rajaganesh a, Mathath Roni a, Al Thabiani Aziz c,
Hatem Ahmed Naji Al-Aoh d, Subrata Trivedi c, Hasibur Rehman c, Suresh Kumar e,
Akon Higuchi f, Angelo Canale g, Giovanni Benelli g, *
a
Division of Entomology, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, 641046, Tamil Nadu, India
b
Thiruvalluvar University, Serkkadu, Vellore 632 115, Tamil Nadu, India
c
Biology Department, Faculty of Science, University of Tabuk, Tabuk, Saudi Arabia
d
Chemistry Department, Faculty of Science, University of Tabuk, Tabuk, Saudi Arabia
e
Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, 43400 Serdang, Slangor, Malaysia
f
Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001 Taiwan
g
Department of Agriculture, Food and Environment, University of Pisa, Via Del Borghetto 80, 56124 Pisa, Italy

a r t i c l e i n f o a b s t r a c t

Article history: The overuse of synthetic pesticides to control insect pests leads to physiological resistance and adverse
Received 23 December 2016 environmental effects, in addition to high operational cost. Insecticides of botanical origin have been
Accepted 2 January 2017 reported as useful for control of agricultural and public health insect pests. This research proposed a
Available online xxx
novel method of mangrove-mediated synthesis of insecticidal silver nanoparticles (AgNP) using Suaeda
maritima, acting as a reducing and stabilizing agent. AgNP were characterized by UVevis spectroscopy,
Keywords:
Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), energy-dispersive
Arbovirus
X-ray spectroscopy (EDX), and X-ray diffraction (XRD) analysis. S. maritima aqueous extract and
Crop protection
Mangrove
mangrove-synthesized AgNP showed larvicidal and pupicidal toxicity against the dengue vector Aedes
Antibacterial activity aegypti and the tobacco cutworm Spodoptera litura. In particular, LC50 of AgNP ranged from 8.668 (larva I)
Nanotechnology to 17.975 ppm (pupa) for A. aegypti, and from 20.937 (larva I) to 46.896 ppm (pupa) for S. litura. In the
field, the application of S. maritima extract and AgNP (10
LC50) led to 100% mosquito larval reduction
after 72 h. Smoke toxicity experiments conducted on A. aegypti adults showed that S. maritima leaf-,
stem- and root-based coils evoked mortality rates comparable or higher if compared to permethrin-
based positive control (62%, 52%, 42%, and 50.2 respectively). In ovicidal experiments, egg hatchability
was reduced by 100% after treatment with 20 ppm of AgNP and 250 ppm of S. maritima extract.
Furthermore, low doses of the AgNP inhibited the growth of Bacillus subtilis, Klebsiella pneumoniae and
Salmonella typhi. Overall, our results highlighted the potential of S. maritima-based herbal coils and green
nanoparticles as biopesticides in the fight against the dengue vector A. aegypti and the tobacco cutworm
S. litura.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction were caused by 20e40% decrease in crop yield, due to the attack
from pathogenic organisms and insect pests [109]. Agriculture is
According to the report of FAO, US $120 billion losses worldwide the backbone of the Indian economy and nearly 75% of the rural
areas of Indian villagers are depending on agriculture. The amount
of food production is greatly deteriorated due to the crop pests and
* Corresponding author. diseases, which lead to agricultural damage either directly by
E-mail addresses: cpselva@ymail.com (C. Panneerselvam), benelli.giovanni@ causing economic losses to the crops in the field or indirectly by
gmail.com (G. Benelli).

http://dx.doi.org/10.1016/j.pmpp.2017.01.002
0885-5765/© 2017 Elsevier Ltd. All rights reserved.

Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
2 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11
causing organoleptic alterations and/or the production of toxic
substances [6].
The tobacco cutworm Spodoptera litura (Fab). (Lepidoptera:
Noctuidae), is one of the major pests of many important crop
plants. Since cutworm larvae can defoliate many economically
important crops possessing a high dispersal capability, this pest
often leads to high levels of agricultural losses [18,27]. This pest
attacks more than 112 species of cultivated crops. Currently, large
quantities of insecticides have been used to fight cutworm in-
festations on different crops [72]. Chemical pesticides play a sig-
nificant role in increasing agricultural production by controlling the
insect pests. Also, molecular research has revealed the interaction
of autophagy-related protein 1 with autophagy-related protein 5 in
S. litura [108]. However, there is widespread concern over negative
impact of insecticides on environmental and human health due to
accumulation of insecticide residues as well as emergence of
pesticide resistance in the pests [5,9]. Due to this reason, many
researchers focused on alternative control methods. Botanicals
are effective against a variety of insect pests; they are easily
degradable [15].
Besides crop pests, mosquitoes represent the major arthropod
vectors of human disease worldwide, transmitting malaria,
lymphatic filariasis, and arboviruses such as dengue fever and Zika
virus [10,11,47,77]. Dengue, mainly vectored by the bites of infected
Aedes mosquitoes, has the greatest epidemic potential worldwide,
with huge negative impact on the economy and health of the
population in urban areas [12e14]. Dengue can be divided into four
serotypes (DENV 1, 2, 3, and 4), each of which confers partial cross-
protective immunity to the other serotypes in humans [99]. Dengue
is affecting more than 128 countries, and the results of biometric
analysis of dengue burden in Arabian countries have been recently
revealed [110]. Ever year about 390 million people are infected by
dengue virus, among which 96 million become severe and results
in about 21,000 deaths [16]. About half of the world's population is
now at risk.
The main transmission cycle is identified for dengue, which is
largely caused by the urban adapted Aedes aegypti mosquitoes, and
along with some other species such as Aedes albopictus [12,74].
Currently, a global alert has been issued for Zika, given the increase
in congenital abnormalities, Guillain-Barre
syndrome, and other
autoimmune manifestations, as well as the increase in chronic joint
diseases due to chikungunya [62].
Since dengue and Zika virus are currently not vaccine pre-
ventable communicable diseases, vector control remains the only
way to prevent arbovirus transmission [13,20,34]. As such, vector
control and personal protection from the bites of infected
mosquitoes are necessary [104]. Effective dengue control requires
the community's participation. The community's health knowl-
edge, attitudes and practices (KAP) will determine their partici-
pation in community-based programs. Taken together, these
scenarios highlighted the need for effective and sustainable vector
control strategies [9]. Very recently it is reported that a particular
strain of Wolbachia can reduce the transmission of Zika virus by
A. aegypti [2]. Overall, new drugs with unique structures and
mechanisms of action are urgently required to treat drug-resistant
strains of dengue [12]. Natural products and their derivatives from
plants are continued to play an important role in the development
of drugs for the treatment of human diseases as well as mosqui-
tocides [64,112].
Mangroves are a rich source of biologically active and pharma-
cologically valuable natural products [40,80]. Therefore the present
research presents recent advances in order to develop insecticidal
compounds from mangrove plant extracts, with special reference to
the green synthesis of silver nanoparticles. Mangrove forest eco-
systems are characterized by facultative halophytic species of trees
Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
and shrubs that fringe the intertidal zone along sheltered coastal,
estuarine and riverine areas in tropical and subtropical latitudes
[8,37]. Mangroves are biochemically unique vegetation that pro-
duces a wide array of natural products with immense medicinal
potential [68,69]. They have been traditionally used in fisher-folk
medicine to treat several diseases.
It has been reported that the mostly halophytic genus Suaeda
consists of 110 species worldwide covering the coastal areas of
tropical and subtropical regions [29]. The distribution and zonation
of different mangrove species also depends on physico-chemical
variations of salinity and available nutrients [48,98]. Overall,
mangroves have an immense ecological role in the coastal and
marine environment [97].
Suaeda maritima (L.) Dumort (Chenopodiaceae) is a salt marsh-
mangrove annual herb that grows in very alkaline and saline moist
soils [78,80]. This plant is distributed throughout the east-west
coast mangroves in India, i.e. Sunderbans in West Bengal, Mah-
nadhi and Bitharkanika in Orissa, Coringa, Krishna and Godavari in
Andhra Pardesh, Karangadu and Pichavaram in Tamil Nadu. Leaf
extracts of S. maritima have been used as traditional remedies for
hepatitis [41], viral [61,71] and bacterial infections [45].
Nanotechnology is a promising field of interdisciplinary
research. It opens up a wide array of opportunities in various
fields including pest control, pharmaceuticals, electronics and
parasitology [52e54,73]. Nowadays, the green synthesis of insec-
ticidal nanoparticles is an interesting issue of nanoscience
[11,25,46,57,75,77,81,84]. Synthesis of silver nanoparticles using
mangroves scarcely analyzed the potential of nano-insecticides for
insect pest management [58].
Therefore, in this research, a selected mangrove species
(S. maritima) was used for biosynthesis of silver nanoparticles
(AgNP) effective against insect pests of medical and agricultural
relevance. S. maritima-fabricated AgNP were characterized by
UVevis spectroscopy, Fourier transform infrared (FTIR) spectros-
copy, scanning electron microscopy (SEM), energy-dispersive X-ray
spectroscopy (EDX), and X-ray diffraction (XRD) analysis. We
investigated the toxicity of the aqueous extract of S. maritima and
S. maritima-synthesized AgNP in laboratory conditions against
larvae and pupae of the dengue vector A. aegypti and the tobacco
cutworm S. litura. We also evaluated the impact of S. maritima
extract and AgNP as ovicides on A. aegypti. The smoke toxicity of
herbal coils prepared using different parts of S. maritima on
A. aegypti adults was studied. Both insecticides were validated in
the field against A. aegypti. Finally, we also assessed antibacterial
properties of AgNP against Bacillus subtilis, Klebsiella pneumoniae,
and Salmonella typhi.
2. Materials and methods
2.1. Suaeda maritima collection and extraction
S. maritima leaves were collected from coastal areas of
Pichavaram (11250 47.900 N 79 480 08.500 E, Cuddalore district),
Tamil Nadu, India. Specimens were washed with tap water and
shade-dried at room temperature. Dried leaves were powdered
using an electrical blender; 500 g of the powdered plant material
were extracted using 1.5 L of ethanol for 72 h. The crude plant
extract was concentrated at reduced temperature using a rotary
evaporator, and stored at 22  C. One gram of the residue was
dissolved in 100 mL of acetone (fixative agent to separate the
aqueous impurities altering the chemical com-position of plant
crude extract) and considered as 1% stock solution. From this
stock solution, experimental concentrations were prepared
[52,55].
 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11
2.2. Aedes aegypti and Spodoptera litura rearing
A. aegypti were reared following the method by Suresh et al.
and Murugan et al. [93,56] in laboratory conditions [27 ± 2  C,
75e85% R.H., 14:10 (L:D) photoperiod]. S. litura larvae were
collected from the Central Institute of Cotton Research, Indian
Council of Agricultural Research, Govt. of India, Coimbatore, India.
They were cultured in laboratory and fed with Ricinius communis
leaves ad libitum at 27 ± 2  C, 75e85% R.H., with 14:10 (L:D)
photoperiod. Pre-pupae of S. litura were separated and provided
with vermiculite clay, which is a good medium for pupation. Pu-
pae of S. litura were kept on cotton in Petri dishes inside an adult
emergence cage. The emerging moths were fed with 10% sucrose
solution fortified with a few drops of vitamin mixture (MULTI DEC
Vitamin drops) to enhance oviposition. Moths in the ratio of one
male to one female were allowed inside oviposition cages con-
taining the adult food mentioned above. The egg cage of S. litura
was covered with white muslin cloth for egg laying. The egg
clothes were removed daily and surface sterilized using 10%
formaldehyde solution to prevent virus infection. The egg clothes
were moistened and kept in a plastic container for the eggs to
hatch. This process facilitated un-interrupted supply of test
insects.
2.3. Biosynthesis and characterization of silver nanoparticles
The S. maritima aqueous leaf extract was prepared adding 10 g
of washed and finely cut leaves in a 300-ml Erlenmeyer flask
filled with 100 ml of sterilized double distilled water and then
boiling the mixture for 5 min, before finally decanting it. The
extract was filtered using Whatman filter paper n. 1, stored
at 4  C and tested within 5 days. The filtrate was treated with
aqueous 1 mM AgNO3 solution in an Erlenmeyer flask and
incubated at room temperature. A brown-yellow solution indi-
cated the formation of AgNP, since aqueous silver ions were
reduced by the S. maritima extract generating stable AgNP in
water. Silver nitrate was purchased from the Precision Scientific
Co. (Coimbatore, India).
Green-synthesis of AgNP was confirmed by sampling the reac-
tion mixture at regular intervals and the absorption maxima was
scanned by UVevis, at the wavelength of 200e550 nm in UV-3600
Shimadzu spectrophotometer at 1 nm resolution. Furthermore, the
reaction mixture was subjected to centrifugation at 15,000 rpm for
20 min, resulting pellet was dissolved in deionized water and
filtered through Millipore filter (0.45 mm). An aliquot of this filtrate
containing AgNP was used for scanning electron microscopy (SEM),
Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction
(XRD) analysis, and energy dispersive X-ray (EDX) spectroscopy.
The structure and composition of freeze-dried purified AgNP was
analyzed by using a 10 kV ultra high-resolution scanning electron
microscope with 25 ml of sample was sputter coated on copper stub
and the images of AgNP were studied using a FEI QUANTA-200
SEM. The surface groups of the AgNP were qualitatively
confirmed by FTIR spectroscopy (Stuart 2002), with spectra recor-
ded by a Perkin-Elmer Spectrum 2000 FTIR spectrophotometer.
EDX assays confirmed the presence of metals in analyzed samples
[55,56].
2.4. Mosquito larvicidal and pupicidal toxicity in laboratory
conditions
Twenty-five A. aegypti larvae (I, II, III or IV instar) or pupae were
placed for 24 h in a glass beaker filled with 250 ml of dechlorinated
water plus the desired concentration of S. maritima leaf extract (50,
100, 150, 200 and 250 ppm) or green-synthesized AgNP (5, 10, 15,
Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
3
20 and 25 ppm). Larval food (0.5 mg) was provided for each tested
concentration [39]. Each concentration was replicated five times
against all instars. Control mosquitoes were exposed for 24 h to the
corresponding concentration of the solvent. Percentage mortality
was calculated as follows:
Percentage mortality ¼ (number of dead individuals/number of
treated individuals)*100
2.5. Mosquito field larvicidal assays
Following the method by Suresh et al. [93]; the S. maritima
leaf extract or AgNP were applied in six external water storage
reservoirs at the National Institute of Communicable Disease
Centre (Coimbatore, India), using a knapsack sprayer (Private
Limited 2008, Ignition Products, India). Pre-treatment and post-
treatment observations were conducted at 24, 48 and 72 h us-
ing a larval dipper. Toxicity was assessed against third- and
fourth-instar larvae. Larvae were counted and identified to spe-
cific level. More than 90% of all surveyed larvae belong to
A. aegypti. Six trials were conducted for each test site with similar
weather conditions (28 ± 2  C; 80% R.H.). The required
quantity of mosquitocidal was calculated on the basis of the total
surface area and volume (0.25 m3 and 250 L); the required
concentration was prepared using 10
LC50 values [51,93]. Per-
centage reduction of the larval density was calculated using the
formula:
Percentage reduction ¼ (C e T)/ C
100
where C is the total number of mosquitoes in the control and T is
the total number of mosquitoes in the treatment.
2.6. Toxicity against the tobacco cutworm Spodoptera litura
Toxicity against S. litura larvae and pupae was studied using the
leaf disk assay with no-choice method. F2 generation larvae were
fed with cotton leaf disks treated with different concentrations of
S. maritima extract and mangrove-synthesized AgNP using the
dipping method. After 24 h, the individuals were transferred to
untreated fresh cotton leaves. The leaves were changed every 24 h.
Mortality was recorded after 96 h of treatment. Five replicates were
maintained for each treatment with 10 larvae per replicate (total,
n ¼ 50). Percentage mortality was calculated using the following
formula [36]:
Corrected mortality ¼ (Mortality in treatment - mortality in con-
trol)/(100-mortality in control)*100.
The survived larvae were fed with untreated cotton leaves until
pupation. Pupal mortality was calculated by subtracting the num-
ber of emerging adults from the total number of pupae.
2.7. Smoke toxicity assays against Aedes aegypti
Leaves of S. maritima were used to prepare herbal coils for
smoke toxicity assays against A. aegypti. Coils were prepared as
described by Suresh et al. [93]; using 4 g of powdered leaves, 2 g of
sawdust (binding material) and 2 g of coconut shell charcoal
powder (burning material). The three materials were mixed with
distilled water forming a semi-solid paste. Mosquito coils (0.6 cm
thickness) were prepared from the semi-solid paste and then dried
in the shade. Negative control coils were prepared following the
4 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11
same method, without adding S. maritima. Positive control was
commercial permethrin-based coils [93].
Following [54], experiments were conducted in a glass chamber
measuring 140 cm
120 cm
60 cm. A door measuring
60 cm
30 cm was situated at the front of the chamber. In each
test, 100 adult female mosquitoes (age: five days old, blood-starved
for three days) were released into the chamber and were provided
with a 10% (w:v) sucrose solution. An immobilized pigeon with a
shaven belly was tied inside the tightly closed chamber. Each pi-
geon was used only once. The experiment was repeated five times
on five separate days for each treatment (i.e. S. maritima-based coil,
positive and negative controls). All mosquitoes were exposed to the
vapor of burning coils for 1 h. After each experiment, the number of
fed and unfed (alive and dead) mosquitoes were counted. The
protection provided by the smoke from the plant samples against
biting A. aegypti was calculated in terms of percentage of unfed
mosquitoes due to treatment:
[(Number of unfed mosquitoes in treatment e Number of unfed
mosquito in negative control) /Number of treated
mosquitoes]
100
2.8. Mosquito ovicidal activity
Following Su and Mulla [90] in ovicidal activity experiments,
A. aegypti eggs were collected placing ovitraps (i.e. Petri dishes,
diameter 60 mm, lined with filter paper and containing 100 ml of
water) inside each cage. In A. aegypti assays, ovitraps were stored in
the cages for 2 days from the blood meal of females. The eggs laid
on filter paper lining were examined using a photomicroscope
(Leica ES2, Germany). Then, the eggs were placed in a cage with six
glass cups (diameter: 6 cm). Five of them were filled with water
plus the S. maritima extract and AgNP treatments as follows: 50,
100, 150, 200 and 250 and 5, 10, 15, 20 and 25 ppm. The control cup
was filled with distilled water. 100 eggs were placed in each cup.
Five replicates were done for each dosage. After treatment, the eggs
from each concentration were transferred to distilled water cups
for hatching assessment after counting the eggs under microscope.
The percent egg mortality was calculated on the basis of non-
hatchability of eggs with unopened opercula [111]. The hatch
rates were assessed 48 h post-treatment using the following for-
mula [31]:
Egg mortality (%) ¼ (number of hatched larvae/total number of
eggs)*100
2.9. Antibacterial activity
S. maritima-synthesized AgNP were tested against B. subtilis,
K. pneumoniae and S. typhi. All bacteria strains were provided by
Microbial Type Culture, Collection and Gene Bank Institute of Mi-
crobial Technology, Sector 39-A, Chandigarh-160036 (India). For all
species, bacterial cultures 18e24 h old were used for the prepara-
tion of testing cultures. All bacteria were grown in nutrient broth
described by Dinesh et al. [22]. Each bacterial strain was inoculated
and incubated at 37  C for 24 h. After this phase, the culture
attained 2
106 cfu/mL, and was used for antibacterial assays. The
antibacterial activity of S. maritima AgNP was assessed using the
agar disk diffusion method [22]. The tested bacteria strains were
swabbed on Muller-Hinton agar medium plates. Three sterilized
filter paper disks, treated with three different concentrations of the
Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
tested compounds, were inserted in each plate. Plates were incu-
bated at 37  C for 24 h. After the incubation, the zone of inhibition
was measured using a photomicroscope (Leica ES2, Germany). The
zone of inhibition indicates the degree of sensitivity of bacteria to a
given treatment; a bigger area of bacteria-free media surrounding
an antibiotic disk means the bacteria are more sensitive to the
compound(s) the disk contains [76].
2.10. Data analysis
SPSS software package 16.0 version was used for all analyses. For
both insect species, acute toxicity data from laboratory assays and
bacteria inhibition growth data were transformed into arcsine/
proportion values and then analyzed using a two-way ANOVA with
two factors (i.e., dosage and mosquito instar or bacterium species).
Means were separated by Tukey's HSD test. Furthermore, insect
pest mortality data from laboratory assays were analyzed by probit
analysis, calculating LC50 and LC90 following the method by Finney
[28]. Mosquito larval density data from field assays were analyzed
using a two-way ANOVA with two factors (i.e. the mosquitocidal
treatment and the elapsed time from treatment). Means were
separated using Tukey's HSD test (P < 0.05). In herbal coil toxicity
experiment, the number of fed, unfed and dead mosquitoes were
analyzed by a one-way ANOVA where the factor was the treatment
(i.e. the coil). Means were separated using Tukey's HSD test
(P < 0.05). Ovicidal data were transformed into arcsine√proportion
values and analyzed by one-way ANOVA. Means were separated
using Tukey's HSD test (P < 0.05).
3. Results and discussion
3.1. Characterization of S. maritima-fabricated silver nanoparticles
The rapid synthesis of AgNPs was observed when the aqueous
extract of S. maritima was incubated with the AgNO3 solution. The
aqueous extract behaved as both reducing (from Agþ to Agо) as well
as capping/stabilizing agent for nascent AgNPs. The rapid change of
coloration indicated the formation of AgNPs. The absorption
spectra of AgNPs at different time intervals showed highly sym-
metric absorption bands. A maximum absorption peak was
observed at 340 nm (Fig. 1). The biosynthesis of AgNP can be
confirmed by the formation of yellowish brown color, and this
might be due to the excitation of the surface Plasmon vibration of
Fig. 1. UVevisualization of the absorption spectra of Suaeda maritima-synthesized Ag
nanoparticles over different time intervals.
 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11 5

the synthesized AgNP [46]. Similarly Sangeetha et al. [82], reported

AgNP synthesized using mangrove Excoecaria agallocha showed
surface Plasmon absorbance bands at 434 nm. Furthermore Uma-
shankari et al. [100], highlighted that the absorption spectrum of
Rhizophora mucronata-synthesized AgNP showed an intense peak
at 426 nm.
XRD patterns showed intense peaks corresponding to (111),
(200) and (220) Bragg's reflection based on the face-centered cubic
structure of AgNP. Thus, XRD highlighted that AgNP formed by the
reduction of AgNO3 with S. maritima leaf extract were crystalline in
nature (Fig. 2) [70,85]. Furthermore Dubey et al. [24], reported the
size of Ag nanocrystals as estimated from the full width at half
maximum of the (111) peak of silver using the Scherrer formula was
20e60 nm. Additionally, the unassigned peaks were identified due
to the presence of phytochemicals from extracts that may be
capping on the surface of AgNP [87].
SEM studies shed light on size and morphological features of
S. maritima-synthesized AgNP. S. maritima-synthesized AgNPs were
predominantly spherical in shape (Fig. 3a). As regard to mangrove-
based synthesis [58], showed Sonneratia alba-synthesized AgNP
were mostly spherical and cubical in shape and showed a large
distribution of sizes, with mean values of 20e60 nm. With refer-
ence to algae [57], reported that the SEM of Centroceras clavulatum-
synthesized AgNP showed spherical and cubic structures with a
size range of 35e65 nm. Furthermore, AgNPs fabricated using the
seaweed frond extract of Caulerpa scalpelliformis were spherical in
shape with an average size ranging from 20 to 35 nm [52] while
Roni et al. [81] reported spherical AgNPs with size ranging from 40
to 65 nm fabricated with Hypnea musciformis extract. EDX revealed
a strong signal in the Ag region, confirming the presence of
elemental silver (Fig. 3b). Metallic silver nanocrystals showed a
typical optical absorption peak approximately at 3 keV due to
surface plasmon resonance [38,102]. EDX also showed the presence
of oxygen, suggesting that AgNP were capped by the organic
components present in the mangrove extract, as also highlighted by
FTIR analyses [67].
FTIR studies were done in order to identify the possible bio-
molecules in S. maritima leaf extract, which may be responsible for
synthesis and stabilization of AgNP (Fig. 4). The FTIR spectrum of
S. maritima leaf extract-fabricated AgNP shows bands at
3525.57 cm1, 3290.34 cm1, 2944.38 cm1, 2831.64 cm1,
1718.49 cm1, 1647.87 cm1, 1407.42 cm1, 1113.32 cm1,
1020.24 cm1 and 596.33 cm1. The presence of peak at 3416 cm1 Fig. 3. (a) Scanning electron microscopy (SEM) micrograph showing the morpholog-
could be ascribed to OeH groups from polyphenols, proteins, en- ical characteristics of silver nanoparticles synthesized using the Suaeda maritima
zymes and/or polysaccharides [94], while the peak at 2922 cm1 extract; (b) Energy dispersive X-ray (EDX) spectrum of silver nanoparticles synthesized
indicates carboxylic acid [43]. The strong intense peak at 1382 cm1 using S. maritima.

probably corresponds to CeN stretch vibrations, as well as to the

amide I bands of proteins in the leaf extract [33]. The band at
1636.40 cm1 corresponds to C]O stretching of alcohols, amide I
and nitro groups [83]. The peak obtained at 1019 cm1 is probably
800
linked to the CeN stretching vibration of aliphatic amines, alcohols
or phenols [105].
600
Intensity (cps)

3.2. Larvicidal and pupicidal toxicity on Aedes aegypti

400
In laboratory assays, the leaf extract of S. maritima was moder-
ately toxic to larval instars (I-IV) and pupae of Ae. aegypti, even if
200
tested at low doses. with LC50 values ranging from 135 to 242 ppm
(Table 1). Mortality was proportional to the tested dosage. Recent
research has showed that different mangrove species can be a
0 source of compounds with good mosquitocidal properties. For
20 40 60 80
instance Yogananth et al. [107], reported that essential oil from
2-theta (deg)
Rhizophora mucronata mangrove leaves was highly toxic to IV instar
Fig. 2. X-ray diffraction pattern of Suaeda maritima-synthesized silver nanoparticles. larvae of An. stephensi and Cx. quinquefasciatus with LC50 values of

Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
6 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11

Fig. 4. Fourier transforms infrared (FTIR) spectrum of vacuum-dried S. maritima-synthesized silver nanoparticles.

Table 1
Acute toxicity of Suaeda maritima leaf extract against larvae and pupae of the dengue vector Aedes aegypti.

Target LC50 (LC90) (ppm) 95% confidence limit LC50 (LC90) Regression equation c2 (df ¼ 4)
LCL UCL

I instar 135.034 (329.083) 116.007 (289.219) 152.403 (393.778) y ¼ 0.892 þ 0.007x 2.334 n.s.
II instar 162.454 (380.995) 142.983 (327.934) 183.844 (473.007) y ¼ 0.953 þ 0.006x 1.249 n.s.
III instar 183.604 (399.344) 164.172 (343.293) 208.032 (496.463) y ¼ 1.091 þ 0.006x 1.816 n.s.
IV instar 210.809 (436.988) 188.381 (370.755) 243.539 (556.311) y ¼ 1.194 þ 0.006x 3.150 n.s.
Pupa 242.949 (468.268) 216.137 (394.569) 286.328 (603.626) y ¼ 1.382 þ 0.006x 2.589 n.s.

LC50 ¼ lethal concentration that kills 50% of the exposed organisms.

LC90 ¼ lethal concentration that kills 90% of the exposed organisms.
c2 ¼ chi-square value.
d.f. ¼ degrees of freedom.
n.s. ¼ not significant (a ¼ 0.05).

0.051 mg/mL (An. stephensi) and 0.0514 mg/mL (Cx. quinquefascia-
tus) [56]. observed mortality in larvae and pupae of Ae. aegypti
exposed to aqueous extract of the mangrove Bruguiera cilyndrica.
Besides Balakrishnan et al. [7], studied the larvicidal activity of the
marine actinobacteria, Streptomyces alboflavus from mangrove
environment, which showed significant activity against Ae. aegypti
(LC50 1.48) and An. stephensi (LC50 1.30). In addition Ravikumar et al.
[80], noted that the mangrove extracts of B. cylindrica, Ceriops
decandra, Lumnitzera racemosa, R. apiculata, and R. mucronata
exerted antiplasmodial activity against chloroquine-sensitive Plas-
modium falciparum. As regards to the toxicity mechanisms, we have
observed that the mangrove extract plus AgNPs can act as in-
hibitors of neurosecretory cells, leading to shirinkage of internal
cuticle, and/or can act directly on epidermal cells responsible for
the production of enzymes leading tanning and/or cuticular
oxidation process [50,86].
S. maritima-synthesized AgNP were highly toxic against Ae.
aegypti. LC50 ranged from 8 to 17 ppm (Table 2). To the best of our

Table 2
Acute toxicity of Suaeda maritima-synthesized silver nanoparticles against larvae and pupae of the dengue vector Aedes aegypti.

Target LC50 (LC90) (ppm) 95% confidence limit LC50 (LC90) Regression equation c2 (df ¼ 4)
LCL UCL

I instar 8.668 (21.407) 0.909 (17.036) 12.302 (34.381) y ¼ 0.872 þ 0.101x 9.487 n.s.
II instar 10.102 (24.322) 8.386 (22.114) 11.512 (27.476) y ¼ 0.910 þ 0.090x 5.178 n.s.
III instar 12.239 (27.460) 10.648 (24.846) 13.645 (31.276) y ¼ 1.031 þ 0.084x 1.946 n.s.
IV instar 14.893 (31.490) 13.354 (28.197) 16.429 (36.482) y ¼ 1.150 þ 0.771x 2.906 n.s.
Pupa 17.975 (35.348) 16.386 (31.358) 19.846 (41.572) y ¼ 1.326 þ 0.074x 4.248 n.s.

LC50 ¼ lethal concentration that kills 50% of the exposed organisms.

LC90 ¼ lethal concentration that kills 90% of the exposed organisms.
c2 ¼ chi-square value.
d.f. ¼ degrees of freedom.
n.s. ¼ not significant (a ¼ 0.05).

Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
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 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11
knowledge, this is the first report about the toxicity of S. maritima-
synthesized nanoparticles against arthropods [58]. have reported
that the Sonneratia alba-synthesized AgNPs were toxic against
larvae and pupae of A. aegypti, LC50 values ranging from 3.15 ppm (I)
to 13.61 ppm (pupa). Another study by Balakrishnan et al. [7]
investigated that the larvicidal activity of AgNP synthesized using
the leaf extract of Avicennia marina against Ae. aegypti and An.
stephensi with LC50 values were 4.374 (An. stephensi) and 7.406 mg/
L (Ae. aegypti). Dinesh Kumar et al. [23] reported that the R.
lamarckii-fabricated AgNP exhibit high HIV type 1 reverse tran-
scriptase inhibitory activity with IC50 of 0.4 mg/ml. Gnanadesigan
et al. [30] highlighted that the R. mucronata-fabricated AgNP tested
against Ae. aegypti and Cx. quinquefasciatus had LC50 values ranging
from 0.585 mg/L (Ae. aegypti) and 0.891 ppm (Cx. quinquefasciatus).
In the field, the application of S. maritima leaf extract or AgNP
(10
LC50) leads to the complete elimination of larval populations
of A. aegypti after 72 h (Table 3). Field assays investigating mos-
quitocidal efficacy of green-synthesized AgNP are currently limited
[22,91]. For example Suresh et al. [93], reported that the field
application of P. niruri extract (10
LC50) lead to Ae. aegypti larval
reduction of 39.9, 69.2, and 100%, after 24, 48, and 72 h, respec-
tively. Rajaganesh et al. [77] highlighted a single application of
Dicranopteris linearis fern extract in water storage reservoirs lead to
100% larval reduction of the dengue vector Ae. aegypti after 72 h.
Also Panneerselvam et al. [65], reported that the leaf extract of
Euphorbia hirta was highly effective in field trials against An. ste-
phensi, as it led to larval density reductions of 13.17, 37.64 and
84.00% after 24, 48 and 72 h.
The biotoxicity of these botanical preparations might be due to
the thin oily layer of constituents with little polarity, which spread
on the water surface cutting oxygen supply to mosquito larvae. In
addition, a number of plant-borne compounds dissolve into the
water and penetrate into the larvae through the respiratory tubes
or cuticle, killing them by suffocation and/or poisoning [53,55,101].
For S. maritima-synthesized AgNP, we hypothesize that the high
mortality rates exerted on Ae. aegypti may be due to the small size
of AgNP, which allows them to pass through the insect cuticle and
even into individual cells, where they interfere with molting and
other physiological processes [10,44,66,91,92]. With reference to Si-
based nanoformulates, it has been reported that the mode of action
for insecticidal activity of nanosilica is through desiccation of insect
cuticle by physicosorption of lipid and is also expected to cause
damage in the cell membrane, resulting in cell lysis and death of the
insects [96]; see also [81].
3.3. Smoke toxicity of S. maritima-based coils
Smoke toxicity experiments conducted on A. aegypti adults
showed that S. maritima leaf-, stem- and root-based coils evoked
mortality rates comparable or higher if compared to permethrin-
based positive control (62%, 52%, 42%, and 50.2 respectively)
(Table 4). In agreement with our experiments, after a single treat-
ment with the leaf-, stem-, and root-based coils prepared using
P. niruri, the percentages of unfed mosquitoes was 58%, 40%, and
61% [93]. [55] also highlighted the concrete potential to produce
mosquitocidal coils against A. stephensi using the seaweed Ulva
Table 3
Field reduction of Aedes aegypti larval populations post-treatment with Suaeda maritima leaf extract and green-synthesized silver nanoparticles in water storage reservoirs.
Suaeda maritima extract (10xLD50)
Before treatment 24 h 48 h
a b
Larval density 122.60 ± 8.32 85.60 ± 5.45 51.20 ± 8.67
Means ± SD followed by different letter(s) are significantly different (ANOVA, Tukey's HSD test, P < 0.05).
Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
7
lactuca as burning material. Furthermore, Vineetha and Murugan
[103] reported that smoke toxicity effects of Aegle marmelos and
Toddalia asiatica against Aedes aegypti. Haldar et al. [35], studied the
effect of smoke toxicity of Ficus krishnae against An. stephensi and
Cx. vishnui. The smoke toxicity against Aedes vectors is probably due
to the presence of active compounds from the different plant parts,
which are toxic for the central nervous system, producing the
knockdown effect [1].
3.4. Ovicidal activity
In ovicidal experiments, egg hatchability of A. aegypti was
reduced by 100% after treatment with 25 and 30 ppm of AgNP; the
S. maritima extract exerted 100% mortality post-treatment
with 250 ppm, while control eggs showed the 100% hatchability
(Table 5). Currently, most botanical preparations, including nano-
formulated ones, focused on larvicidal assays, while the ovicidal
potential, which usually require higher doses to be effective, re-
mains overlooked [10]. In this framework Rajaganesh et al. [77],
pointed out the ovicidal activity of D. linearis-fabricated AgNP
against Ae. aegypti, while Madhiyazhagan et al. [111] observed that
the Sargassum muticum-synthesized AgNP was an effective ovipo-
sition deterrent against An. stephensi, Ae. aegypti and Cx. quinque-
fasciatus at 50 ppm. Later on, Chandramohan et al. [19] have
reported that the ovicidal properties Acorus calamus-synthesized
AgNP on the malaria vector Anopheles stephensi. Concerning plant
extracts, Cocculus hirsutus methanol extract caused 86% and 100%
ovicidal activity against An. subpictus when tested at 500 ppm and
1000 ppm [26], while Govindarajan and Rajeswary [32] observed
that the methanolic leaf extract of Albizia lebbeck exerted zero
hatchability on Cx. quinquefasciatus, Ae. aegypti, and An. stephensi at
250, 200, and 150 ppm, respectively. Recently Munusamy et al. [49],
showed that the methanol extract of Rubia cordifolia root had good
ovicidal activity (82.40% and 70.40%) against the eggs of Cx. quin-
quefasciatus and Ae. aegypti, at 500 mg/L when compared to other
plants, such as Gymnema sylvestre, Scilla peruviana, S. cordifolia and
Elytraria acaulis (see also [4,64].
3.5. Toxicity against the tobacco cutworm Spodoptera litura
The S. maritima leaf extract was moderately toxic to larvae and
pupae of Spodoptera litura. LC50 ranged from 268 to 609 ppm
(Table 6). To the best of our knowledge, studies on mangrove-based
biopesticides and related compounds against agricultural pests,
with special reference to the tobacco cutworm, are scarce. Rani
et al. [79] studied antifeedant and toxic activity of Hibiscus tiliaceus
and Sonneratia caseolaris on S. litura. Furthermore Deshmukhe et al.
[21], reported that the aqueous crude extract of Lantana camara
leaves achieved 100% mortality on fourth instar larvae of S. litura
when tested at 40%. Ananthi and Ranjitha Kumari [3] studied that
the larvicidal activity of seed and root extract of Rorippa indica
against S. litura. In addition to acute toxicity, a long term mortality
effect evoked by mangrove extract on S. litura larvae may be due the
active plant compounds entering into the body of the larvae and
suppressing the activity of ecdysone, thus larva fails to molt,
leading to disruption of normal physiological and metabolic
Green-synthesized Ag nanoparticles (10xLD50)
72 h Before treatment 24 h 48 h 72 h
cd e a c d
0.00 ± 0.00 135.40 ± 6.76 59.00 ± 8.63 37.80 ± 6.14 0.00 ± 0.00e
8 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11

Table 4
Smoke toxicity assays conducted using Suaeda maritima-based coils against the biting activity of the dengue vector Aedes aegypti.

Treatment Fed mosquitoes (%) Unfed mosquitoes (%) Total (%)

Alive Dead

Leaf-based coil 14.4 ± 1.67b 24.6 ± 2.40a 62.00 ± 2.12d 86.6 ± 0.54d
Stem-based coil 21.8 ± 0.83c 28.4 ± 1.67ab 51.8 ± 1.09c 79.8 ± 1.92c
Root-based coil 28.4 ± 1.14d 31.6 ± 1.34b 41.6 ± 1.51b 72.2 ± 0.83b
Negative control (blank coil) 75.2 ± 1.48e 25.0 ± 1.58a 0.0 ± 0.0a 25.0 ± 1.22a
Positive control (pyrethrin-based coil) 10.20 ± 1.30a 40.6 ± 1.67c 50.2 ± 1.48c 90.2 ± 1.92d

Within each column, different letters indicate significant differences (ANOVA, Tukey's HSD, P < 0.05).

Table 5
Ovicidal activity of Suaeda maritima leaf extract and Suaeda maritima-synthesized silver nanoparticles on Aedes aegypti.

Treatment Egg hatchability (%)

Control 50 ppm 100 ppm 150 ppm 200 ppm 250 ppm
a b c d e
Suaeda maritima extract 86.2.83 ± 0.83 61.40 ± 1.14 41.80 ± 1.30 29.20 ± 0.83 21.2 ± 1.30 NH
Ag nanoparticles Control 5 ppm 10 ppm 15 ppm 20 ppm 25 ppm
91.20 ± 0.83a 53.00 ± 1.58b 36.6 ± 1.14c 25.2 ± 1.30d NH NH

Values were means ± SD of 5 replicates.

Within a row, different letters indicated significant differences (ANOVA, TUkey's HSD, P < 0.05).
NH ¼ no hatchability.

processes [59].
The S. maritima-synthesized AgNP were more effective than the
mangrove extract alone on S. litura. LC50 ranged from 20 to 46 ppm
(Table 7). To the best of our knowledge, most toxicity studies on
green synthesized nanoparticles focused on pests of medical and
veterinary importance (mainly mosquitoes and ticks), while efforts
on crop pests are rather rare. Durga Devi et al. [25] reported that the
E. hirta-fabricated AgNP showed good insecticidal properties
against Helicoverpa armigera, with LC50 values ranging from
2.905 ppm (I) to 3.086 ppm (pupa). Furthermore, Roni et al. [81],
highlighted H. musciformis-synthesized AgNP were highly toxic
against P. xylostella, LC50 were 24.51 ppm (I), 26.47 ppm (II),

Table 6
Acute toxicity of Suaeda maritima leaf extract on larvae and pupae of Spodoptera litura.

Target LC50 (LC90) (ppm) 95% confidence limit LC50 (LC90) Regression equation c2 (df ¼ 4)
LCL UCL

I instar 268.784 (631.237) 233.137 (560.088) 301.371 (742.857) y ¼ 0.950 þ 0.004x 0.171 n.s.
II instar 310.383 (696.523) 275.329 (611.553) 346.622 (834.415) y ¼ 1.030 þ 0.003x 0.018 n.s.
III instar 348.772 (757.492) 312.319 (657.766) 391.528 (925.063) y ¼ 1.094 þ 0.003x 0.077 n.s.
IV instar 393.165 (815.147) 353.620 (701.541) 445.957 (1011.14) y ¼ 1.194 þ 0.003x 0.150 n.s.
V instar 448.269 (874.430) 402.646 (746.680) 516.636 (1100.09) y ¼ 1.348 þ 0.003x 0.375 n.s.
VI instar 530.823 (945.397) 473.002 (801.036) 626.767 (1207.19) y ¼ 1.641 þ 0.003x 0.172 n.s.
Pupa 609.045 (992.761) 537.923 (836.771) 734.968 (1284.33) y ¼ 2.034 þ 0.003x 0.408 n.s.

LC50 ¼ lethal concentration that kills 50% of the exposed organisms.

LC90 ¼ lethal concentration that kills 90% of the exposed organisms.
c2 ¼ chi-square value.
d.f. ¼ degrees of freedom.
n.s. ¼ not significant (a ¼ 0.05).

Table 7
Acute toxicity of Suaeda maritima-synthesized silver nanoparticles on larvae and pupae of Spodoptera litura.

Target LC50 (LC90) (ppm) 95% confidence limit LC50 (LC90) Regression equation c2 (df ¼ 4)
LCL UCL

I instar 20.937 (50.663) 17.438 (45.907) 23.824 (57.557) y ¼ 0.903 þ 0.043x 4.094 n.s.
II instar 23.936 (54.790) 20.659 (49.496) 26.792 (62.566) y ¼ 0.994 þ 0.042x 3.770 n.s.
III instar 26.981 (61.987) 23.558 (55.216) 30.140 (72.455) y ¼ 0.988 þ 0.037x 1.672 n.s.
IV instar 31.798 (67.300) 28.586 (59.713) 35.193 (79.158) y ¼ 1.148 þ 0.036x 1.706 n.s.
V instar 35.961 (73.483) 32.561 (64.526) 40.020 (87.956) y ¼ 1.228 þ 1.034x 1.900 n.s.
VI instar 39.238 (77.648) 35.597 (67.771) 43.965 (93.901) y ¼ 1.309 þ 0.033x 1.550 n.s.
Pupa 46.896 (85.347) 42.440 (73.825) 53.521 (104.862) y ¼ 1.563 þ 0.33x 0.558 n.s.

LC50 ¼ lethal concentration that kills 50% of the exposed organisms.

LC90 ¼ lethal concentration that kills 90% of the exposed organisms.
c2 ¼ chi-square value.
d.f. ¼ degrees of freedom.
n.s. ¼ not significant (a ¼ 0.05).

Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
 U. Suresh et al. / Physiological and Molecular Plant Pathology xxx (2017) 1e11
Table 8
Zone of inhibition induced by Suaeda maritima-synthesized silver nanoparticles on
the bacteria B. subtilis, K. pneumoniae and S. typhi.
Species Inhibition zone (mm)
Control 30 ppm 60 ppm 90 ppm
a b c
B. subtilis 8.16 ± 0.15 16.10 ± 0.10 20.16 ± 0.15 23.10 ± 0.10d
K. pneumoniae 9.03 ± 0.05a 14.20 ± 0.17b 17.03 ± 0.05c 21.1 ± 0.20d
S. typhi 9.83 ± 0.15a 12.06 ± 0.11b 18.13 ± 0.11c 20.0 ± 0.03d
Values are means ± standard deviation of 3 replicates.
Within a row, different letters indicate significant differences (ANOVA, Tukey's HSD,
P < 0.05).
28.35 ppm (III), 32.55 ppm (IV) and 38.23 ppm (pupa). Yasur and
Usha Rani [106] studied the impact of AgNP on growth and feeding
responses of two lepidopteran pests, S. litura and Achaea janata. The
insecticidal properties of green fabricated nanoparticles are mainly
due to their morphological features, in particular their optical
properties coupled with the high surface:volume ratio, that allow a
boosted delivery of toxic bioactive compounds from plants, leading
to key physiological changes [60]; it is worthy to note that their use
is new in the field of agricultural pest management [10e13,17].
3.6. Antibacterial activity
In our experiments, S. maritima-synthesized AgNPs showed
good antibacterial properties against B. subtilis, K. pneumoniae, and
S. typhi (Table 8). At the maximum concentration tested (90 ppm),
AgNP showed strong inhibitory action against B. subtilis (zone of
inhibition 23.10 mm), K. pneumoniae (zone of inhibition 21.1 mm)
and S. typhi (zone of inhibition 20.0 mm) (Table 8). Currently, green
synthesized nanoparticles have been extensively surveyed against
microbial pathogens. As a recent example related to mangroves
[58], showed the good antibacterial activity of S. alba-synthesized
AgNPs against B. subtilis, K. pneumoniae, and S. typhi. Furthermore
Thatoi et al. [95], reported that the Heritiera fomes and S. apetala-
synthesized AgNP showed high antibacterial potential than the ZnO
NP against Staphylococcus aureus, Shigella flexneri, Vibrio cholera,
Staphylococcus epidermidis, Bacillus subtilis, and Escherichia coli.
Panja et al. [63]. reported that the antimicrobial activity of Rauvolfia
serpentina-fabricated AgNP was high on B. subtilis, Enterococcus
faecalis, Pseudomonas aeruginosa, and E. coli, using disk methods. In
addition, Mahyoub et al. [46] highlighted that AgNP fabricated
using Halodule uninervis showed good antibacterial activity against
B. subtilis, K. pneumoniae and S. typhi.
The exact mechanism for the antibacterial activity of AgNPs is
not fully clarified. However, many studies report that the AgNP
could bind to the bacterial membrane, invade the cell and cause
appetite of proton motive force which leads to the distruction of
bacterial cell by forming pores on the cell wall. Secondly, AgNP can
interfere with the thiol group of bacterial cells leading to ceasure of
respiratory chain reaction, cell division and death [89].
4. Conclusions
We have reported an easy, simple, and environmentally friendly
approach for the synthesis of AgNP mediated by mangrove leaf
extract of S. maritima as a reducing and capping/stabilizing agent.
UVeVis spectrophotometry, XRD analysis, SEM, EDX spectroscopy,
and FTIR spectroscopy confirmed the rapid and cheap synthesis of
AgNP. Our results pointed out that S. maritima and seaweed-
synthesized AgNP toxic against invertebrates of medical and agri-
cultural importance, including dengue mosquitoes and the tobacco
cutworm, allowing us to propose the tested products as effective
candidates to develop newer and safer botanical insecticides. We
Please cite this article in press as: U. Suresh, et al., Suaeda maritima-based herbal coils and green nanoparticles as potential biopesticides against
the dengue vector Aedes aegypti and the tobacco cutworm Spodoptera litura, Physiological and Molecular Plant Pathology (2017), http://
dx.doi.org/10.1016/j.pmpp.2017.01.002
9
also noted that relatively low doses of green-synthesized AgNP
effectively inhibit several species of microbial pathogens [46].
Further studies are in progress to optimize this synthesis route,
evaluating the toxicity of intermediate reaction products on
different insect pests of economic relevance.
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