Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62

Contents lists available at ScienceDirect

Egyptian Journal of Basic and Applied Sciences

journal homepage: www.elsevier.com/locate/ejbas

Full Length Article

A comparative study on larvicidal potential of selected medicinal plants

over green synthesized silver nano particles
Syed Zameer Ahmed Khader a, Sidhra Syed Zameer Ahmed a,⇑, Jagadeeswari Sathyan a,
Mohamed Rafi Mahboob b, Kisore P. Venkatesh a, Kishore Ramesh a
a
Department of Biotechnology, K.S. Rangasamy College of Technology, K.S.R. Kalvinagar, Tiruchengode 637 215, Tamil Nadu, India
b
Department of Science & Humanities, Kingston Engineering College, Katpadi, Vellore, Tamil Nadu 632059, India

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

Article history: Larvicidal activity was assessed for alcoholic extracts of Phyllanthus amarus, Annona squamosa, Coccinia
Received 26 September 2017 grandis and Eclipta prostrata extracted using solvents of various polarity. Third instar stage larvae of
Received in revised form 8 January 2018 Dengue-vector, Aedes aegypti and Japanese encephalitis (JE) causing mosquito Culex tritaeniorhynchus
Accepted 8 January 2018
were subjected to larvicidal bioassay at various concentrations (1000, 500, 250 ppm). The results
Available online 1 February 2018
explored that the phytoconstituents and secondary metabolites present in all the plants elucidated
potent larvicidal activity. Among the tested extract ethyl acetate, petroleum ether and hexane extract
Keywords:
expressed significant larvicidal activity. Similarly, these plants were subjected to green synthesis of silver
Larvicidal
Silver nanoparticle
nanoparticles, characterized and subjected for its larvicidal activity against Anopheles stephensi causing
Green synthesis malaria. The synthesized silver nanoparticles were characterized by UV–VIS spectroscopy, Fourier
Folklore Transform Infra-Red spectroscopy, Scanning Electron Microscopy respectively. The FTIR analysis strongly
supported the capping behaviour of bio-reduced synthesized silver nanoparticles which in turn imparted
the high stability of the synthesized silver nanoparticles. The average size of synthesized nanoparticles
was less than 1 mm, most spherical in shape with SEM analysis. The findings revealed that Eclipta prostrata
and Annona squamosa has effective larvicidal activity, whereas all the synthesised nanoparticles demon-
strated dose dependent activity even at very low concentration and the findings reveals that these
extracts and nanoparticles can be a better remedy against these mosquitoes.
Ó 2018 Mansoura University. Production and hosting by Elsevier B.V. This is an open access article under
the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction control breeding of Aedes aegypti as it is a fresh water breeding

mosquito.
Mosquito borne diseases is a major universal health problem Whereas JE (Japanese encephalitis) is caused by mosquito
causing malaria, dengue fever, Japanese encephalitis, chikungunya vector Culex tritaeniorhynchus distributed mostly in south Asia
fever and lymphatic filariasis which are more prevalent in India. It and southeast Asia [5] and has reported a mortality of about
has been reported that 2.5 million cases have been reported with 30,000–50,000 annually. These vectors breeds especially in irriga-
malaria and India alone contributed 76% of the cases [1]. Anopheles tion waters of rice field and uses pigs as their amplification host.
stephensi is the malaria causing mosquito of plasmodium genus. The symptoms of Japanese Encephalitis infections are mild, with
About 400 known Anopheles species are normally found or without apparent symptoms, but many reports suggest that cer-
everywhere except cold temperate regions. Anopheles stephensi is tain patients results in severe disease characterized by raise in
a major insect vector in the world has been shown directly respon- body temperature, headache, neck stiffness, disorientation, coma,
sible for about 40–50% of the annual malarial incidence [2]. Simi- seizures, spastic paralysis and death. Control of mosquito is some-
larly, Aedes aegypti a vector of Dengue fever has created a serious thing important in the present day with rise in number of illness
public health problem globally. Among the arbovirus in India, caused by mosquito and it is a major sources for death in develop-
spreading all the dengue virus types is constantly increasing [3]. ing countries.
It has been reported that around 50–200 million cases of Dengue Bioactive compounds from plant origin plays a major role as a
fever infection worldwide every year [4]. It is difficult task to defence agent against various insects. A diversity of plants have
been reported with insecticidal compounds and the use of
⇑ Corresponding author. products of plants origin to control mosquito larvae has been
E-mail address: [email protected] (S. Syed Zameer Ahmed). shown to be an exciting alternative to traditional methods of larval

https://doi.org/10.1016/j.ejbas.2018.01.002
2314-808X/Ó 2018 Mansoura University. Production and hosting by Elsevier B.V.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62 55

controlling, as they are not associated with the problems [6]. Sev- methods. They also find place in the insecticides like
eral research has been carried out to make eco-friendly repellents nanomaterials-based insecticides. They provide green and efficient
and larvicidal activity using plants [7,8]. Medicinal plants are alternatives for pest management without harming the nature. The
found to be the alternative medicine used since ancient times to atom by atom arrangement of nanoparticle influences their size,
cure many diseases due to their therapeutic values. Medicinal shape and orientation to act specifically on the targeted site. The
plants have many characteristics which make them useful for nanoparticle possess more strength, chemical reactivity and a high
treatments, they are synergic medicine, supportive medicine and electrical conductivity, along with their physical, biological and
preventive medicine [9]. chemical properties.
Nanotechnology finds application in different areas like health It has been reported that Silver nanoparticles possess anti-
care, cosmetics, drug-gene delivery, energy science, optoelectron- bacterial activity [12] similarly, antiviral [13], anti-fungal activity
ics, photo-catalyst property [10,11] etc. Nanoparticles provides [14]. Nano particles synthesized from microorganism or plants
solutions to technological and environmental problems and also sources acts can potentially to eliminate the problem by making
provides high surface area to volume ratio. Green synthesis of the nanoparticles more bio-compatible, hence many researchers
nanoparticles helps to develop a clean, nontoxic, and eco-friendly are working on plants, algae, fungi, bacteria, and viruses to produce

Fig. 1. FTIR spectrum of synthesized silver nanoparticles using extracts of Phyllan- Fig. 3. FTIR spectrum of synthesized silver nanoparticles using extracts of Eclipta
thus amarus. prostrata.

Fig. 2. (a-c) SEM micrograph a) 1 500 10 mm; B) 5000 5 mm; C) 10,000 1 mm showing the silver nanoparticles synthesized using extract of Phyllanthus amarus.
56 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62

low-cost, energy-efficient, and nontoxic production of metallic 2.3. Preparation of extract

nanoparticles [15].
The silver nanoparticle efficacy depends on the particle size and Dried powdered samples of Phyllanthus amarus, Annona squa-
their shape. The previous studies shows that reducing agents like mosa, Coccinia grandis and Eclipta prostrata were taken separately
borohydride gave smaller monodisperse silver nanoparticles. The and mixed with different polar and non-polar and then magneti-
use of less reducing agent like citrate gave a slower reducing rate. cally stirred in a separate container for overnight at room temper-
The chemical synthesis of nanoparticle resulted in toxic and less ature. The residue was removed by filtration. The filtrate was
effective nanoparticles. Green synthesis of nanoparticle is concentrated under reduced pressure in a rotary evaporator at 6
eco-friendly and less toxic. The green synthesis of nanoparticle 0 ± 10 °C to yield concentrated crude extract and the extracts were
depends on the selection of substances which are nontoxic, stored at 4 °C for future use without deterioration [16].
solvents and reducing agent for stability of silver nanoparticles.
The experiment is designed to validate the applications of folklore
medicine to control the prevalence of diseases. 2.4. Synthesis of silver Nano particle

AgNPs were synthesized from the whole plant material (Phyl-

2. Materials and method lanthus amarus, Annona squamosa, Coccinia grandis and Eclipta pros-
trata) by the method explained by Rajakumar and Rahuman, [17].
2.1. Collection and processing of plant material

The whole plant samples of Phyllanthus amarus, Annona squa-

mosa, Coccinia grandis and Eclipta prostrata were collected from
bottom of Kolli Hills, Namakkal District. These scientific names of
these plants were morphologically identified using the database
and classified based on their family and authenticated. The fresh
leaves were collected, washed and shade dried. The leaves were
coarsely powdered by using electric blender and stored separately
in an air tight container for further use.

2.2. Chemicals and solvents

All the chemicals and solvents were of analytical grade and

obtained from S.D. Fine Chemicals, Mumbai and Fischer Inorganic Fig. 5. FTIR spectrum of synthesized silver nanoparticles using extracts of Annona
and Aromatic Limited, Chennai, India. squamosa.

Fig. 4. (a-c) SEM micrograph a) 1500 10 mm; B) 5000 5 mm; C) 10,000 1 mm showing the silver nanoparticles synthesized using extract of Eclipta prostrata.
 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62 57

Fig. 6. (a-c) SEM micrograph a) 1500 10 mm; B) 5000 5 mm; C) 10,000 1 mm showing the silver nanoparticles synthesized using extract of Annona Squamosa.

2.5. Collection of mosquito larvae

The third instar stage larvae of Anopheles stephensi, Aedesaegypti

and Culex tritaeniorhynchus were collected from Medical Entomo-
logical Centre, ICMR (field station), Virudhachalam, Cuddalore
District. The larvae were maintained in aplastic tray containing
water with sufficient feed. The tray was covered with muslin cloth
to avoid contamination. The larvae were provided with suitable
environmental condition of 25–27 °C and 75–85% humidity and
the egg rafts were collected from each cage for maintaining next
generation.

2.6. Bioassay for larvicidal activity Fig. 7. FTIR spectrum of synthesized silver nanoparticles using extracts of Coccinia
grandis.
The assay was carried out according to guidelines of WHO
2.7. Dose-response bioassay
(1996), larvae are added to 249 ml of water and 1.0 ml of prepared
plant extracts by maintaining five replicates. The control will be
The synthesized Nano particles were subjected to larvicidal
maintained without extract and the replicates are left for exposure
activity against larvae of Anopheles stephensi following the method
for 24 h. Then the number of viable larvae will be reported for
of Rajakumar and Rahuman, 2011 [18].
assessment of mortality rate from the mean of triplicates. The
extracts showing higher activity will be considered for further
studies. 3. Results
Percentage mortality = (No of dead larva/No of larva intro
duced)  100 3.1. Characterization of silver nanoparticles
Study involves 5 larvae for each sample. The stock solution
(1000 ppm) was prepared by combination of 100 ml of distilled Synthesis of silver nanoparticles was confirmed by the values and
water, 0.4 ml of acetone, 100 mg of extract and 0.02 ml of Tween from characterization of the particle using UV, FTIR and SEM
20 (for complete dissolution of extract). By diluting further, solu- analysis.
tions of concentration 1000, 500, 250 ppm was obtained. Negative
control was maintained for all the samples containing only solvent 3.1.1. FTIR and SEM analysis of Phyllanthus amarus
without extract. A commercial liquidator was used as positive con- FTIR measurement was carried out to identify the possible
trol at three concentrations. biomolecules responsible for capping and efficient stabilization of
58 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62

Ag nanoparticle synthesized by plant leaf extract. FTIR analysis image showed the particle size of about 1 mm as well the crystal
showed the presence of different functional groups corresponding structure of the nanoparticles.
to proteins, alkaloids, tannins, saponins and other plant metabo-
lites. The FT-IR spectrum of AgNPs from P. amarus is shown in 3.1.3. FTIR and SEM analysis of Annona squamosa
Fig. 1. The characteristics absorbance bands due to aldehydic The FT-IR spectrum of AgNPs from Annona squamosa is shown in
CAH stretch at 2930 cm 1. The band at 1638 cm 1 is due to CAO Fig. 5. The peak at 3428 cm 1 is due to the NAH group of amines.
stretch and the band at 1401 cm 1 is for NAO stretch of amines. The C@O and CAH group are appeared at 1627 cm 1 of esters and
The band at 1326 cm 1 is due to the CAO stretch of dialkyl, and alkenes. The NAH and CAH group is appeared at 1411 cm 1 is for
1049 cm 1 is for the CAN. The band at 614 cm 1 is due to aromatic amines and alkanes. The C@O for esters are appeared at 1050 cm 1.
CAH stretch. The broad band which appeared at 3436 cm 1 is the The C„C group appeared at 566 cm 1 is for alkynes.
characteristic absorption for OAHAO of water. The SEM image showing the high intensity of silver nanoparti-
The SEM of silver nanoparticles of P. amarus which showed the cles synthesized by Annona squamosa extract further confirmed the
synthesis of monodisperse spherical Ag NPs (Fig. 2). The SEM development of silver nanostructures. The synthesized Ag NPs
image showing silver nanoparticles synthesized using plant extract were monodispersed, spherical in shape with size range between
confirmed the development of silver nanostructures by P. amarus. 100 and 200 nm and they are quite well distributed without any
The shape of Ag NPs was spherical and size was less than 300 agglomeration (Fig. 6). The synthesized nanoparticles were fine
nm synthesized from P. amarus. stabilized by capping agent (plant phytochemicals) thus they were
not in direct contact even within the aggregates as seen in SEM
3.1.2. FTIR and SEM analysis of Eclipta prostrata image.
The FT-IR spectrum of AgNPs from Eclipta prostrata is shown in
Fig. 3. The band at 3385 cm 1 for Hydroxy group, H-bonded OH 3.1.4. FTIR and SEM analysis of Coccinia grandis
stretch. The band at 1610 cm 1 was assigned to amide. The peak The FT-IR spectrum of AgNPs from Coccinia grandis is shown in
at 1415 cm 1 which expose symmetric stretching of COO from Fig. 7. The peaks observed for Ag nanoparticles formed through
an amino group. The peak at 1049 cm 1 is for Phosphate ion and reduction by C. grandis, at 1090 cm 1 (ether linkages), 1404 cm 1
the peak at 617 cm 1 for Alcohol. The IR band proved the presence (@NH) and 1639 cm 1 (amide I) suggest the existence of ter-
of amines, alcohols, phenols, and aromatic groups and strongly penoids and alkaloids attached on the surface of silver nanoparti-
proposes the presence of certain proteins in Eclipta prostrata that cles. The analysis of IR spectrum also provided an idea about
act as reducing/capping agents and may be responsible for the syn- biomolecules containing different functional groups which are pre-
thesis of AgNPs using leaves of E. Prostrata. sent in the underlying system.
SEM analysis provided further detailed insight into the mor- The micrograph showed the particle size of about 200 nm as
phology and size details of the silver nanoparticles. The SEM con- well the crystal structure of the nanoparticles, with uniformly dis-
firmed the development of silver nanostructures and gave clear tributed silver nanoparticles on the surface of the cells is observed.
image of silver nanoparticles from Eclipta Prostrata (Fig. 4). The However, it does not indicate that all the nanoparticles are

Fig. 8. (a-c) SEM micrograph a) 1500 10 mm; B) 5000 5 mm; C) 10,000 1 mm showing the silver nanoparticles synthesized using extract of Coccinia grandis.
 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62 59

3.1.5. X-ray diffraction analysis

The XRD patterns of AgNPs with Phyllanthus amarus, Eclipta
prostrata, Annona squamosal, Coccinia grandis are shown in Fig. 9
(a-d). The diffraction image shows two peaks appeared with 2h val-
ues and these peaks are corresponding to the (1 1 1) and (2 0 0)
planes of face cantered cubic structure of silver which are well
matched with the standard card (JCPDS 89–3722). The unexpected
peaks could be due to the crystallisation of bioorganic phase that
occurs on the surface of the AgNPs.

3.2. Larvicidal bioassay of terrestrial plants against Aedes aegypti

The larval mortality of Phyllanthus amarus, Annona squamosa,

Eclipta prostrata and Coccinia grandis against Aedes aegypti are given
in Fig. 10. Analysis with Phyllanthus amarus demonstrated a maxi-
mum activity of 46.66% and 39.33% of mortality in petroleum ether
and benzene extracts at 1000 ppm, whereas hexane extract repre-
sented minimum activity of 2.6%. Similarly, Hexane extract of E.
prostrata posed maximum 100% activity at 500 ppm and ethyl
Fig. 9. (a-d) XRD pattern of synthesized AgNPs (a-Phyllanthus amarus, b-Eclipta
prostrata, c-Annona Squamosa, d-Coccinia grandis) exhibiting the facets of crystalline acetate extract at 1000 ppm represented 93.33%. Its minimum
silver. activity was observed in methanol and water extracts of about
6.66%. Meanwhile, petroleum ether and ethyl acetate extracts of
Annona squamosa illustrated maximum of 66.66 and 73.33% of
attached to the surface of the cells, since those dispersing in the mortality respectively. Its water and benzene extracts showed
solution may also deposit onto the surface of the cells. The SEM minimal activity of 13.33%. Only the methanolic leaf extract of C.
results (Fig. 8) represented that synthesized AgNPs were cubic in grandis showed 33.33% mortality at 1000 ppm. The plant showed
shape with 300–1 mm size and found to be well dispersed in aque- no activity at 250 and 500 ppm concentration of hexane, methanol
ous medium of Coccina grandis extract. and benzene extracts.

Fig. 10. Mortality rate of Aedes aegypti larvae against terrestrial plant extracts.
60 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62

Fig. 11. Mortality rate of Culex tritaeniorhynchus larvae against terrestrial plant extracts.

Fig. 12. Larvicidal activity of synthesized Ag NPs against Anopheles stephensi.

 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62 61

Table 1 reduction of Ag+ ions with leaf extract are predominantly spherical
Larvicidal activity of synthesized AgNPs against Anopheles stephensi. in shape they are quite well distributed without any agglomeration
Nanoparticles LC50 (UCL-LCL) LC90(UCL-LCL) v2 (df = 5) [9]. At low magnification SEM image shows monodispersed NPs
Pa AgNPs 2.819 (3.362–2.447) 6.204 (10.798–4.731) 0.331 with the size ranging from 200 to 300 nm.
As AgNPs 2.332 (2.809–1.937) 6.279 (12.545–4.569) 1.813 The XRD patterns clearly demonstrates that the AgNPs formed
Ep AgNPs 2.735 (3.246–2.370) 6.291 (10.307–4.648) 2.256 in the current synthesis were crystalline in nature. The sharpening
Cg AgNPs 2.509 (2.967–2.152) 5.843 (10.014–4.461) 3.881 of the peaks clearly specifies that the particles are in the nanore-
Control – nil mortality. Significant at P < .05 level. LC50: lethal concentration that gime. Nanoparticle XRD patterns exhibit several different size-
kills 50% of the exposed larvae, LC90 lethal concentration that kills 90% of the dependent features leading to anomalous peak positions, intensity
exposed larvae, UCL upper confidence limit, LCL lower confidence limit, v2 chi- and widths. Appearances of these peaks are due to the presence of
square; df degree of freedom.
phytochemical compounds in the extracts. The stronger planes
indicate Ag NPs as a key element in the biosynthesis. The minor
3.3. Larvicidal bioassay of terrestrial plants against Culex shift in the peak positions in all the samples indicated the presence
tritaeniorhynchus of some strain present in the crystal structure [21].

The larval mortality of Phyllanthus amarus, Annona squamosa,

Eclipta prostrata and Coccinia grandis against Culex tritaeniorhynchus
4.2. Larvicidal activity
are given in Fig. 11. Analysis demonstrates that Phyllanthus amarus
was highly effective with 100% mortality in ethyl acetate and hex-
It is evident that there is a global alarm is in raise against vector
ane extracts at 1000 ppm concentration, but petroleum ether
control due to the development of resistance in vector mosquitoes
extract revealed around 93.33% mortality. Its water and methanol
over conventional insecticides. Hence the present study focuses on
extract explicated lowest activity around 6.6 to 13%. E. prostrata
the development of larvicides using medicinal plants like Phyllan-
showed 100% mortality in petroleum ether, ethyl acetate, hexane
thus amarus, Annona squamosa, Coccinia grandis and Eclipta pros-
and methanol extracts at 250, 500 & 1000 ppm. Its benzene and
trata and to validate its traditional usage. An attempt is made to
water extract explicated minimum activity of 26% and 33.3%
green synthesize silver nanoparticles from whole plant and to
respectively. Annona squamosa exhibited 100% mortality in petro-
identify its lethal effect over mosquito larva.
leum ether, ethyl acetate and hexane extracts. Meanwhile its ben-
zene and methanol extracts were moderately active with mortality
rate of 80% and 53% respectively. Coccinia grandis demonstrated
100% mortality in petroleum ether extract and 86.66% mortality 4.2.1. Larvicidal activity of terrestrial plants against Aedes aegypti
in benzene extract. Benzene extract showed moderate activity of Larvicidal activity was evaluated against Aedes aegypti and the
66%. Its water and ethyl acetate extract showed minimum activity results revealed that ethyl acetate and petroleum ether extracts
of 20% at 1000 ppm concentration. of all the terrestrial plants were effective. The Eclipta prostrata
extract exhibited higher mortality when ethyl acetate, petroleum
3.4. Larvicidal bioassay for different concentration of nanoparticle ether and hexane were used as solvents. Annona squamosa demon-
against Anopheles stephensi strated does dependent activity against Aedes larvae. Only petro-
leum ether and ethyl acetate extracts of Phyllanthus amarus
Illustration from Fig. 12 demonstrates larvicidal activity of syn- explicated considerable activity. The least activity on Aedes species
thesized Nanoparticles against Anopheles stephensi. Exposure of was seen in Coccinia grandis. Our study is in line with the previous
Phyllanthus amarus nanoparticles at 4 ppm concentration pre- study representing larvicidal activity using medicinal plants [22].
sented 73% mortality where as 3 ppm demonstrated 53% activity, Also it was found that the extracts demonstrated concentration
similarly Annona squamosa based nanoparticle explicated effective dependent activity, ie. increase in mortality rate with increase in
activity of 80% mortality at 4 ppm concentration and 60% mortality concentration of extract. The results are in coincidence with the
at 3 ppm. Eclipta prostrata at 4 ppm exposed groups represented previous reports exploring dose dependent activity supported by
73% mortality and at 3 ppm showed 52% activity. Meanwhile Coc- the presence of phytochemicals which have insecticidal property
cinia grandis NPs at a dose of 4 ppm showed a maximum activity [22,23].
with 83%, and 52% at 3 ppm exposure. All the tested NPs demon-
strated potent larvicidal activity even at lower concentration and
the results are represented in Table 1. 4.2.2. Larvicidal bioassay of terrestrial plants against Culex
tritaeniorhynchus
4. Discussions During the trail with Culex larvae the petroleum ether, ethyl
acetate extracts of the terrestrial plants were effective. The highest
4.1. Characterization of silver nanoparticles activity was observed in petroleum ether, ethyl acetate, hexane
and methanol extracts of E. prostrata. Similarly, Annona squamosa
IR spectroscopy study has confirmed that the carbonyl group of and Phyllanthus amarus ethyl acetate and petroleum ether extracts
amino acid residues and peptides of proteins has a stronger ability represented effective mortality against Culex larvae. Previous
to bind metal, so that the proteins could most possibly form a coat reports suggest that the biological activity of plant extracts might
covering the metal nanoparticles (i.e. capping of Ag NP) to prevent be due to the presence of various compounds, including phenolics,
the agglomeration of the particles, and thus, the nanoparticles are terpenoids, and alkaloids present in plants [24]. The finding of the
stabilized in the medium [19]. present study revealed that these phytoconstituents present in the
A small dip was obtained at 450–500 nm showing the absorp- plant extracts has represented pronounced effect over mosquito
tion spectra of Ag NPs synthesized by Phyllanthus amarus. FTIR con- larvae. Among the tested plant extract Eclipta prostrata and Annona
firms the presence of different functional groups absorb squamosa demonstrated effective larvicidal activity against Culex
characteristic frequencies of IR radiations [20]. larvae and are quite comparable with previous reports of Raheli
SEM analysis which constitutes a large number of uniform sized et al., [25] representing larvicidal activities of leaf extracts of
nanoparticles revealed that the Ag nanoparticles produced by Indigofera arrecta.
62 S.Z.A. Khader et al. / Egyptian Journal of Basic and Applied Sciences 5 (2018) 54–62

4.3. Larvicidal activity of nanoparticle extract of terrestrial plants fruit peel aqueous extract of Annona squamosa and its compounds against
blood-feeding parasites. Parasitol Res 2012;111:2189–99. doi: https://doi.org/
against Anopheles stephensi
10.1007/s00436-011-2671-2.
[8] MohanaRoopan S, Elango G. Exploitation of Cocos nucifera a non-food toward
The results of morality rate of Anopheles stephensi at different the biological and nanobiotechnology field. Ind Crops Prod 2015;67:130–6.
concentration of nanoparticle extracts were illustrated in Fig. 6. [9] Rasool Hassan BA. Medicinal plants (importance and uses). Pharma Anal Acta
2012;3(10):e139.
Coccinia grandis AgNPs represented maximum mortality rate [10] Goudarzi Mojgan, Salavati-Niasari Masoud, Ghanbari Davood, Ahmadi Amin.
against Anopheles stephensi whereas, Annona squamosa AgNPs Synthesis and characterization of Al(OH) nanoparticles and polymeric
explicted activity moderate than Phyllanthus amarus and Eclipta nanocomposites. J Clust Sci 2016;27:25–38. doi: https://doi.org/10.1007/
s10876-015-0895-5.
prostrata AgNPs. The results demonstrated that maximum mortal- [11] Goudarzi Mojgan, Mousavi-Kamazani Mehdi, Salavati Niasari Masoud. Zinc
ity even at very low concentration of 4 ppm. oxide nanoparticles: solvent-free synthesis, characterization and application
Researchers has reported that synthesized AgNPs may have sig- as heterogeneous nanocatalyst for photodegradation of dye from aqueous
phase. J Mater Sci Mater Electron 2017. doi: https://doi.org/10.1007/s10854-
nificant impact on malaria causing vectors and our results are in 017-6560-z.
line with the previous results [17,18,26] similarly, Velayutham [12] Sathishkumar M, Sneha K, Won SW, Cho CWS, Kim Yun YS. Cinnamon
et al., [27] reported silver nanoparticles synthesized using bark zeylanicum bark extract and powder mediated green synthesis of nano-
crystalline silver particles and its bactericidal activity. Colloids Surf. B:
aqueous extract of F. racemosa represented maximum larvicidal Biointerfaces 2009;73:332–8.
activity against the larvae of disease spreading vectors which is [13] Rogers JV, Parkinson CV, Choi YW, Speshock JL, Hussain SM. A preliminary
agrement with our study. assessment of silver nanoparticle inhibition of monkey pox virus plaque
formation. Nanoscale Res Lett 2008;3(4):129–33.
[14] Panacek A, Kolar M, Vecerova R, Prucek R, Soukupova J, Krystof V, et al.
5. Conclusion Antifungal activity of silver nanoparticles against Candida spp. Biomater
2009;30:6333–40.
[15] Thakkar KN, Mhatre SS, Parikh RY. Biological synthesis of metallic
The results of our present study reveals that the folklore medici- nanoparticles. Nanomed Nanotechnol Biol Med 2009;6:257–62.
nes have synergistic effect and can also be used for various ailment [16] Syed Zameer Ahmed K, Sidhra S, Senthil Kumar B, Thanga Kumar A, Geetha K,
without any side effects. Among the tested plants, all plants Mohamed Rafi M, et al. Modulatory effect of dianthrone rich alcoholic flower
extract of Cassia auriculata L. on experimental diabetes. Integr Med Res
expressed activity in different solvents may be due to the presence 2017;6:131–40.
of phytoconstituents in them. Similarly usage of plant based Nano [17] Rajakumar G, Gomathi T, Rahuman AA, Thiruvengadm M, Mydhili G, Kim
materials represented dose dependent mortality even at a lower Seung-Hyun, et al. Biosynthesis and biomedical applications of gold
nanoparticles using Eclipta prostrata leaf extract. Appl Sci 2016;6:222.
concentration and hence it would be an advantage for the develop-
[18] Rajakumar G, Rahuman AA. Larvicidal activity of synthesized silver
ment of clean, nontoxic and environmentally acceptable ‘‘green nanoparticles using Eclipta prostrata leaf extract against filariasis and malaria
approach” to produce metal nanoparticles for betterment of life vectors. Acta Trop 2011;118:196–203.
[19] Rajeswari A, Sujatha D, Balasaraswathi K, Uthirappan Mani, Rose Chellan,
and can stop spreading of adverse diseases caused by mosquito
Mandal Asit Baran. Phytosynthesis of silver nanoparticles using Coccinia
larvae. grandis leaf extract and its application in the photocatalytic degradation.
Colloids Surfaces B Biointerfaces 2012;94:226–30.
Acknowledgement [20] Kalainila P, Subha V, Ernest Ravindran RS, Sahadevan R. Synthesis and
characterization of silver nanoparticles from Erythrina indica. Asian J Pharm
and Clin Res 2014;7:39–43.
Authors are thankful to the Management and Principal of K.S. [21] Khushboo Singh, Manju P, Sangeeta K, Uma C, Jaya Parkash Y. Green silver
Rangasamy College of Technology, Tiruchengode for providing nanoparticles of Phyllanthus amarus: as an antibacterial agent against multi
drug resistant clinical isolates of Pseudomonas aeruginosa. J Nanobiotechnol
infrastructure and facilities to carry out this research work 2014; 12:40. doi: 10.1186/s12951-014-0040-x.
successfully. [22] Baranitharan M, Dhanasekaran S, Mahesh Babu S, Sridhar N, Krishnappa K.
Larvicidal activity of Coleus aromaticus Benth (Lamiaceae) leaf extracts against
three vector mosquitoes. Int J Res Biol Sci 2014;4(2):55–9.
References [23] Pedro G, Aubrey A, Bryle E. Larvicidal activity of selected plant extract against
Dengue vector Aedes aegypti mosquito international research. J Biol Sci 2014;3
[1] Kumar A, Valecha N, Jain T, Dash AP. Burden of malaria in India: retrospective (4):23–32.
and prospective. Am J Trop Med Hyg 2007;77(6):69–78. [24] Rafael L, Teresinha N, Moritz JC, Maria IG, Eduardo MD, Tania SF. Evaluation of
[2] Bashir A, Javid A. Evaluation of larvicidal activity of Hippophae rhammoides L. antimicrobial and antiplatelet aggregation effect of Solidago chilensis eyen. Int J
Leaves extracts on Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). Green Pharm 2009;3:35–9.
Middle-East. J Sci Res 2013;13(5):703–9. doi: https://doi.org/10.5829/idosi. [25] Raheli Neema M, Ojunga M, Ramesh F. The larvicidal activity of Indigofera
mejsr.2013.13.5.2939. arrecta leaf extract against culex mosquito larvae. Int J Bioassays 2015;4
[3] Fulmali PV, Walimbe A, Mahadev PV. Spread, establishment and prevalence of (01):3666–9.
dengue vector Aedes aegypti (L.) in Konkan region, Maharashtra, India. Indian J [26] Santhosh Kumar T, Rahuman, AA, Rajakumar G, Marimuthu S, Bagavan A,
Med Res 2008;127:589–601. Jayaseelan C, Zahir AA, et al., Synthesis of silver nanoparticles using Nelumbo
[4] Murray NE, Quam MB, Wilder Smith A. Epidemiology of dengue: past, present nucifera leaf extract and its larvicidal activity against malaria and filariasis
and future prospects. Clin Epidemiol 2013;5(1):299–309. vectors. Parasitol Res 2011; 108(3): 693–702. doi:10.1007/s00436-010-2115-
[5] Keiser J, Maltese MF, Erlanger TE, Bos R, Tanner M, Singer BH, et al. Effect of 4.
irrigated rice agriculture on Japanese encephalitis, including challenges and [27] Velayutham K, Rahuman AA, Rajakumar G, Mohana Roopan S, Elango G,
opportunities integrated vector management. Acta Trop 2005;95(1):40–57. Kamaraj C, et al. Larvicidal activity of green synthesized silver nanoparticles
[6] Borase HP, Patil CD, Salunkhe RB, Narkhede CP, Salunke BK, Patil SV. Phyto- using bark aqueous extract of Ficus racemosa against Culex quinquefasciatus
synthesized silver nanoparticles: a potent mosquito biolarvicidal Agent. J and Culex gelidus. Asian Pacific J Trop Med 2013;6:95–101.
Nanomedine Biotherapeutic Discov 2013;3:111.
[7] Madhumitha G, Rajakumar G, Mohana Roopan S, Abdul Rahuman A, Mohana
Priya K, Mary Saral A, et al. Acaricidal, insecticidal, and larvicidal efficacy of