International Journal of Nanotechnology and Applications

ISSN 0973-631X Volume 11, Number 1 (2017), pp. 17-28

© Research India Publications

http://www.ripublication.com

Mosquito larvicidal activity of green silver

nanoparticle synthesized from extract of bud of

Polianthus tuberosa L.

Anjali Rawani*

Assistant Professor, Department of Zoology, Laboratory of parasitology, vector biology and nanotechnology,

The University of Gour Banga, Mokdumpur, Malda, 732103, West Bengal , India .

Abstract

Phyto-synthesis of transition metal silver nanoparticles is gaining importance

due to their biocompatibility, low toxicity, green approach and environmental

friendly nature. In the present study, silver nanoparticle synthesized from bud

of Polianthus tuberosa. The reduction of silver ions occurred when the silver

nitrate solution was treated with aqueous extract of bud of Polianthus

tuberosa at 600 C. Synthesized silver nanoparticles (AgNPs) particles were

characterized by analyzing the excitation of surface plasmon resonance (SPR)

using UV–vis spectrophotometer at 450 nm. Further TEM analysis confirmed

the range of particle size as 50±2 nm and nanoparticles are spherical (or oval)

in shape. The XRD spectrum showed the characteristic Bragg peaks which

confirms that these nanoparticles are crystalline in nature. Fourier transform

infrared spectroscopy (FTIR) of synthesized nanoparticles confirms the

presence of amide, phosphate compound, ketones, alkenes, nitro compounds,

aromatic compounds and alkyl compounds. Synthesized Ag nanoparticles

were effective larvicidal agents against Culex vishnui and Culex quinquefasciatus. In larvicidal bioassay with synthesized AgNPs, highest

mortality was observed at 20 ppm against Cx. vishnui with LC50 and LC90

values of 8.25 and 17.99 ppm; 7.46 and 23.26 ppm against 3rd and 4th instars

respectively. While in Cx. quinquefasciatus the LC50 and LC90 values are 9.65

and 27.18; 7.94 and 22. 47 ppm against 3rd and 4th instars respectively. Non

18 Anjali Rawani

target organisms were also exposed to respective lethal concentrations (to

mosquito larvae) of dry nanoparticles which was tested against Toxorhynchites

larvae (mosquito predator), Diplonychus annulatum (predatory water-bug) and

there is no abnormality seen in the non target organisms. These results suggest

that the synthesized AgNPs of P. tuberose have the potential to be used as a

supreme environmental friendly compound for the control of the mosquito

larvae.

Keywords: Polianthus tuberosa, silver nanoparticles, larvicidal activity, non-

target, Probit analysis

INTRODUCTION

Mosquito borne diseases are a serious threat to modern civilization in terms of

mortality, morbidity and economic loss. Among various types of mosquito borne

diseases, Filariasis, Malaria and Dengue fever are most common. Filariasis is caused

by Wuchereria species and mainly transmitted by Culex quinquefasciatus in tropical

regions. A current estimate reveals that about 120 million people in 83 countries of

the world are infected with LF parasites. In India, it is estimated that there are

approximately 21 million people with symptomatic filariasis and 27 million

microfilaria carriers (Reddy et al. 2000). The principle vector of Japanese encephalitis

in tropics is Culex vishnui carrying JEV (An arthrovirus) as causative organism.

Among various ways available to minimize the incidence of mosquito borne diseases

and microbial infections; application of biological products is recommended now a

day as a widely accepted alternative strategy. Biological control achieved through

application of natural product can be long lasting, inexpensive and harmless to living

organisms and ecosystem. It neither eliminates pathogen nor disease, but brings them

into natural balance (Ramanathan et al. 2002).

The field of nanotechnology is now a day considered as an active area of research in

materials sciences (Jahn 1999). Chemically or biologically synthesized nanoparticles

play an important role in pharmaceutical, industrial and biotechnological application

(Saxena et al. 2010; Elumalai et al. 2010). Silver nanoparticles are used as fastest

growing materials due to their unique physical, chemical and biological properties;

small size and high specific surface area. Application of silver nanoparticles, are

reported for their antifungal, anti-inflammatory, antiviral and larvicidal activities

(Rogers et al. 2008; Saxena et al. 2010; Elumalai et al. 2010; Haldar et al. 2013;

Rawani et al. 2013). Green synthesis of silver nanoparticles is favoured over chemical

and physical method as it is cost effective, eco-friendly and also non toxic material

(Lok et al. 2007).

The common name derives from the Latin tuberosa, meaning swollen or tuberous in

reference to its root system. This plant is commonly called ‘Rajanigandha’ in India,

Mosquito larvicidal activity of green silver nanoparticle synthesized from extract 19

which means 'fragrant at night'. It grows up to 45 cm (18 in) long as elongated spikes

that produce clusters of fragrant waxy white flowers that bloom from the bottom

towards the top of the spike. It has long, bright green leaves clustered at the base of

the plant and smaller, clasping leaves along the stem. In the flower epiphyllous

adhesion of stamens is seen. Since the plant are used as essential oil having a calming,

refreshing and sedative aroma. Due to this property the oil is used for the treatment of

emotional stress, anxiety, and sleep disorders. Flowers and bulb are diuretic in nature

which is externally used for skin eruptions. The bulbs have some medicinal properties

and usually rubbed with turmeric and butter and applied over red pimples of infants.

In gonorrhea dried and powdered bulbs are used. The larvicidal and bitting detergency

property of this plant against Anopheles stephensi and Cx. quinquefasciatus mosquito

(Rawani et al., 2011) was also reported.

The objective of this study was to produce silver nanoparticle from fresh bud of

Polianthes tuberosa and analyze the larvicidal activity against two important mosquito

species; Cx. vishnui and Cx. quinquefasciatus as well as to characterize the

synthesized nanoparticles.

MATERIAL AND METHODS

Chemicals used

Silver nitrate (AgNO3) crystal extra pure (Merck), normal saline, double distilled

water, and de-mineralized water were used throughout the experiments.

Plant Materials

Buds of Polianthes tuberosa were collected randomly from gardens of Mokdumpur,

Malda, West Bengal.

Collection of larvae

Raft of Cx. quinquefasciatus eggs were collected from cemented drains surrounding

the University of Gour Banga campus. After hatching, first instar larvae were fed with

small amount of flour. Cx. vishnui larvae were collected from the clear water near rice

field or small ponds containing clear water. Third and fourth instar larvae of both the

species were used in experiment.

Synthesis of silver nanoparticles

After collection from the garden, fresh mature samples of P. tuberosa were washed

repeatedly in laboratory with distilled water to remove any organic impurities present

on it. 10 g of fresh buds crushed to small pieces with a grinder. Pieces were then taken

20 Anjali Rawani

in to 500 mL beaker containing 100mL double distilled water and boiled for 10

minutes. After that the extract was filtered with Whatman filter paper No.42. The

resultant filtrate was used for the reduction of Ag+ to Ag0 (Bhat et al. 2011). The

aqueous extract was treated with aqueous 10-3 M AgNO3 solution and boiled. The

change of color takes place within few minutes from colorless to reddish brown. The

final nano-colloidal solution was subjected to repeated centrifugation (twice) to get

rid of any un-interacted biological molecules at 10,000 rpm for 15 minutes in a Remi

Research Centrifuge instrument. The final pellet was collected, dried in a rotary

evaporator and stored in desiccators for future use. From the synthesized NPs

different test concentrations of aqueous solutions were prepared (10, 15 and 20 ppm)

by mixing with distilled water.

Mosquito larvicidal bioassay

Standard methods for testing biologically synthesized nanoparticle toxicity and the

susceptibility of mosquito larvae to insecticides was performed according to WHO

methodologies (WHO 1996). The larvicidal bioassay was performed at room

temperature. At each tested concentration, four trials were made with a control set up

(having only distilled water). Randomly, twenty 3rd and 4th instar larvae were placed

into 200 ml of each tested concentrations along with control set up and kept in an

environmental chamber at 27°C with a photoperiod of 16:8-h light/dark cycle. The

effectiveness of silver nanoparticles as mosquito larvicides was determined from all

the twenty 3rd and 4th instar larvae with exposure to time periods. The larval

mortalities were assessed to determine the acute toxicities on instar larvae of Cx. Vishnui and Cx. quinquefasciatus after 24 hours of exposure. The numbers of dead

larvae were counted and the percentage of mortality was reported from the average of

four replicates. The larval mortality data were corrected for control mortality by

Abbott’s formula (Abbott, 1925).

Characterization

The formation of Silver nanoparticles was verified by using UV-Vis

spectrophotometer operated at 1nm resolution with an optical length of 10 mm within

190–700 nm wavelength range. UV-Vis analysis of the reaction mixture was observed

for time duration of 300 sec. (Upstone 2000).

For the study of crystallinity, X-Ray-diffraction (XRD) studies were conducted using

Siemens X-Ray diffractrometer (Japan), operated at 30 kV and 20mA current with

CuKα (I=1.54Ao). Films of colloidal form AgNP were tested by drop coating on Si

(III) substrates and data were recorded. The scanning range during the

characterization was selected between 10o to 80o (Hemant et al.2012).

The Transmission Electron Microscopy (TEM) images were obtained using Technai-

20 Philips instrument operated at 200 kV and beam current of 104.1 μA. Sample for

Mosquito larvicidal activity of green silver nanoparticle synthesized from extract 21

this analysis were prepared by coating the aqueous AgNP on carbon coated copper

grids (300 mesh size) by slow evaporation and then allowed to dry in vacuum at 25oC

for overnight (Haldar et al. 2013).

For FTIR studies, the powder sample of AgNP was prepared by centrifuging the

synthesized AgNP solution at 10,000 rpm for 20 min. The solid residue obtained was

washed with deionized water to remove any unattached biological moieties to the

surface of the nano particles. The resultant residue was then dried completely and the

powder obtained was used for FTIR measurements by a FT-IR Spectrometer (Perkin

Elmer Lx10 – 8873). The scanning range was 40 to 4000 cm-1 at a resolution

of 4 cm-1.

Toxicity test on non-target organism

The effect of the nanoparticles of fresh bud of P. tuberosa was tested against non-

target organisms like Diplonychus annulatum and Toxorhynchites larvae as they share

the common habitats of target mosquito larvae. The non-targets were exposed to

lethal concentration (to mosquito larvae) of dry nanoparticles at 24 h (for 3rd instar)

larvae to observe the mortality and other abnormalities such as sluggishness and

reduced swimming activity up to 72 h of exposure.

STATISTICAL ANALYSIS

The data on the efficacy of Ag-NP were subjected to probit analysis (Finney 1971).

By using the computer software "STAT PLUS 2009 (Trial version)" and MS EXCEL

2007 the LC50 and LC90 values, regression equations (Y = mortality; X =

concentrations) and regression coefficient values were calculated.

RESULT AND DISCUSSION

UV-Vis spectroscopy study

The gradual formation of AgNP occurs by bioreduction of Ag+ ions through P. tuberose fresh bud extract. The UV absorption spectrum of silver nanoparticles as a

function of reaction time is shown in Figure 1. The nanoparticle formation kinetics

was monitored by periodic sampling of extract diluted to 3 ml (30 time dilution) by

deionized water taken in a 10-mm-optical path length quartz cuvette. The appearance

of colour and then subsequent changes in colour as a function of time occur due to

initial synthesis of AgNP and subsequent increase in concentration, expressed as

increase in absorbance intensity. The characteristic surface plasmon absorption

spectral band showed at (λmax) 450 nm.

22 Anjali Rawani

Fig 1: Time dependent absorption spectra of Ag NP of fresh bud

TEM Analysis

TEM analysis was done to visualize the shape as well as to measure the diameter of

the Ag nanoclusters. The study reveals that most of the nano-crystals formed are

spherical (or oval) in shape. Figure 2(a), (b) shows a representative TEM photograph

of the synthesized Ag NP from fresh bud of P. tuberosa. Halder et al., 2013 reported

that the major population of the silver nanoparticles is found within 10 to 35 nm range

and the average size of the particles has been calculated to be 26.6 nm. The study also

reveals that most of the nano-crystals formed are quasi-spherical (or polyhedral) in

shape. Rawani et al., 2013 also reported the The nanoparticles are spherical to

polyhedral in shape with size of 50–100 nm (average size of 56.6 nm).

Fig 2a Fig 2b

Fig: 2 (a), (b) TEM image of synthesized silver nanoparticles from bud of P. tuberosa

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

30 min 1 hour 2 hour 3 hour 4 hour 5 hour 6 hour

Absorbance

Mosquito larvicidal activity of green silver nanoparticle synthesized from extract 23

X-ray Diffraction analysis

The dried sample obtained from the ultra-centrifuged AgNP solution was analysed by

the XRD pattern. It showed agreement with the literature values of fcc crystal

structure of Silver. Fig 3 showed the XRD pattern observed from the synthesized

nanoparticles of fresh bud of P. tuberosa. There were intense silver nanoparticle

(AgNP) diffraction peaks at 20.7, 30.3, 40.6 and 50.7 corresponding to facets 111,

200, 220, 311. However, other XRD reports of silver nanoparticles (Bar et al. 2009,

Das et al. 2009, Nirmala et al. 2010, Raghunandan et al. 2010) are in general

agreement with the just-cited results.

Fig 3: XRD of silver nanoparticles from bud of P. tuberosa

FTIR study

FTIR spectroscopy is useful technique to study the core-shell morphology of AgNP

where the absorption of IR radiation takes place through resonance of non-centro

symmetric (IR active) modes of vibration. FTIR absorption spectra of the water

dispersed pellet of the purified Ag NP are tested for FTIR analysis. The interpretation

of the band is given below in Table1.

24 Anjali Rawani

Table 1: FTIR analyses, probable functional group of NPs of fresh bud of P. tuberose

Plant Part Probable Functional Groups

Fresh bud Amide

Phosphate compound

Ketones

Nitro compound

Aromatic compounds/Alkenes

Alkyl compounds

Mosquito Larvicidal Bioassay

The nanoparticles synthesize from fresh bud of P. tuberose were subjected to

larvicidal bioassay separately on third and fourth instars larvae of Cx. vishnui and Cx. quinquefasciatus. Table 2 represents the results of the rate of mortality of 3rd and 4th

instar larvae of Cx. vishnui and Cx. quinquefasciatus when they were exposed to

graded concentrations (10, 15, 20 ppm) of aqueous solution of purified pellet of Ag

NP for 24 hours. The 20 ppm concentration caused 80 % and 76 % mortality in 3rd

instar of Cx. vishnui and Cx. quinquefasciatus respectively. While 88 % and 82.68 %

mortality in 4th instar Cx. vishnui and Cx. quinquefasciatus at 20 ppm respectively

which is highest than 10 and 15 ppm concentration. With decreasing the concentration

of nanoparticle, the mortality rate in all form of larva decreased.

Table 3 represents the LC50 and LC90 values, regression equations along with R values

of Cx. vishnui and Cx. quinquefasciatus larval stages (3rd and 4th) against synthesized

Ag NPs. A clear dose-dependent mortality was observed from the established

regression equations, which is evident from the positive correlation between the rate

of mortality (Y) and the concentration (X) of the Ag NP synthesized.

The data obtained from the present study clearly indicate that silver nanoparticles

could provide excellent larval control of Cx. vishnui and Cx. quinquefasciatus.

Greater mortality is seen in 4th instar larvae than the 3rd instar larvae. The exact

mechanisms of larvicidal effect of AgNPs are yet not known. But it can be assumed

that the AgNPs penetrate through the larval membrane which leads to the death of the

delicate larvae. Marimuthu et al., (2011) reported the antiparasitic activities against

the larvae of malaria vector, An. subpictus Grassi, filariasis vector Culex quinquefasciatus Say (Diptera: Culicidae), and Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae) from the synthesized Ag NPs utilizing aqueous leaf

extract of Mimosa pudica Gaertn (Mimosaceae). Priyadharshini et al., (2012) studied

the larvicidal activity of synthesized Ag NPs utilizing Euphorbia hirta leaf extract

Mosquito larvicidal activity of green silver nanoparticle synthesized from extract 25

against malarial vector An. stephensi. Sareen et al., (2012) reported that the control of

larvae of Aedes albopictus by Ag NPs synthesized from aqueous leaf extract of

Hibiscus rosasinensis. Suman et al., (2013) synthesized Ag NP from aqueous aerial

extract of Ammannia baccifera and applied against An. subpictus and Cx. quinquefasciatus larvae and found that it can effectively inhibit the population of

larvae. Naik et al., (2014) showed moderate larvicidal effects of the synthesized Ag

NPs of leaf of Pongamia pinnata for mosquito control and found that plant extracts

was found to be toxic to larvae at LC50 = 0.25 ppm and LC90 =1 ppm. Rawani et al.

2013 also reported the larvicidal activities of the synthesized AgNPs from fresh

leaves, dry leaves and green berries of S. nigrum against larvae of Culex quinquefasciatus and Anopheles stephensi. Haldar et al.(2013) also showed the

mosquito larvicidal bioassay with the Ag NPs synthesized by dried green fruits of

Drypetes roxburghii against two mosquitoes namely Culex quinquefasciatus and

Anopheles stephensi.

Table 2: Larvicidal activity of synthesized silver nanoparticles using P. tuberose fresh bud extract against 3rd and 4th instars larvae of Cx. vishnui and Cx.

quinquefasciatus

Larval

instar

Mosquito species Concentration

(ppm)

Mean mortality ± SE

3rd instar Cx. vishnui 10 58.68 ± 0.88

15 70.68 ± 0.88

20 80.00 ± 0.58

Cx. quinquefasciatus 10 54.68 ± 0.88

15 64.00 ± 0.58

20 76.00 ± 0.58

4th instar Cx. vishnui 10 64.00 ± 1.15

15 76.00 ± 1.15

20 88.00 ± 0.58

Cx. quinquefasciatus 10 64.00 ± 0.58

15 72.00 ± 1.15

20 82.68 ± 0.67

26 Anjali Rawani

Table 3: Probit analysis and Regression analysis of larvicidal activity of synthesized

silver nanoparticles using P. tuberose fresh bud extract against 3rd and 4th instars

larvae of Cx. vishnui and Cx. quinquefasciatus

Larval

instar

Mosquito species Plant part LC50 LC90 Regression equation R value

3rd instar Cx. vishnui Fresh bud 8.25 17.99 Y=0.77x+8.39 0.94

Cx. quiquefasciatus Fresh bud 9.65 27.18 Y=0.77x+5.50 0.92

4th instar Cx. vishnui Fresh bud 7.46 23.26 Y=0.60x+10.00 0.87

Cx. quiquefasciatus Fresh bud 7.94 22.47 Y=0.67x+8.89 0.89

CONCLUSION

In conclusion, Ag nanoparticles prepared by the herbal origin are cost-effective

biogenic molecules that have great promise as larvicidal agents. From the present

study we conclude that the aqueous extract of fresh bud of P. tuberosa can be used as

an effective capping as well as reducing agent for the synthesis of silver nanoparticles.

Further research is needed to improve the efficiency of production of silver

nanoparticles using P. tuberosa bud extract as a reducing and stabilizing agent and to

increase the biological activity of the product against mosquito larvae to levels

appropriate with commercial product. The mechanisms of mode of action are also

need to study to combat with the effective vector control programme. Based on these

findings, applications of Ag nanoparticles may lead to valuable discoveries in various

fields such as nanomedicine, mosquito control and antimicrobial systems.

REFERENCES

[1] Reddy, G.S., Venkatesvarlou, N., Das, P.K., Vanamail, P., Vijayan, S.K.,

Pani, S.P., 2000, “Tolerability and efficacy of single dose diethylcarbamazine

(DEC) or ivenmectin the clearance of Wuchereria bancrofti microfilaraemia

at Pondicherry, south India”, Trop. Med. Int. Health, 5, pp. 779–785.

doi: 10.1046/j.1365-3156.2000.00644.x

[2] Ramanathan, A., Shanmugam, V., Rghuchander, T., Samiyappan, R., 2002,

“Induction of systematic resistance in ragi against blast diseases by

Pseudomonas flourescens”, Ann. PC Port Soc., 10, pp. 313-318

[3] Jahn, W., 1999, “Chemical aspects of the use of gold clusters in structural

biology”, J Struct Biol 127: 106–112

[4] Saxena, A., Tripathi, R.M., Singh, R.P., 2010, “Biological synthesis of silver

nanoparticles using onion (Allium cepa) extract and their antibacterial

activity”, Dig. J. Nanomater Bios., 5, pp. 427–432

Mosquito larvicidal activity of green silver nanoparticle synthesized from extract 27

[5] Elumalai, E.K., Prasad, T.N.V.K.V., Hemachandran, J., Therasa, S.V.,

Thirumalai, T., David, E., 2010, “Extracellular synthesis of silver

nanoparticles using leaves of Euphorbia hirta and their antibacterial

activities”, J. Pharm Sci. Res., 2(9), pp. 549–554

[6] Rogers, J.V., Parkinson, C.V., Choi, Y.W., Speshock, J.L., Hussain, S.M.,

2008 “A preliminary assessment of silver Nanoparticles inhibition of monkey

pox virus plaque formation”, Nanoscale Res. Lett., 3, pp. 129-133.

doi:10.1007/s11671- 008-9128-2

[7] Haldar, K.M., Haldar, B., Chandra, G., 2013, “Fabrication, characterization

and mosquito larvicidal bioassay of silver nanoparticles synthesized from

aqueous fruit extract of putranjiva, Drypetes roxburghii (Wall)”, Parasitol.

Res., 112(4), pp. 1451–1459. doi: 10.1007/s00436-013-3288-4.

[8] Rawani, A., Ghosh, A., Chandra, G., 2013, “Mosquito larvicidal and

antimicrobial activity of synthesized nano crystalline silver particles using

leaves and green berry extract of Solanum nigrum L. (Solanaceae: Solanales)”

Acta Trop., 128(3), pp. 613– 622. doi: 10.1016/j.actatropica.2013.09.007

[9] Lok, C., Ho, C., Chen, R., He, Q., Yu, W., Sun, H., 2007, “Silver

nanoparticles: partial oxidation and antibacterial activities”, J. Biol. Inorg.

Chem., 12(4), pp. 527-534

[10] Rawani, A., Banerjee, A., Chandra, G., 2012, “Mosquito larvicidal and biting

deterrency activity of bud of Polianthes tuberosa plants extract against

Anopheles stephensi and Culex quinquefasciatus”, Asian Pac. J. Trop. Dis.,

pp. 200‐204

[11] Bhat, R., Deshpande, R., Ganachari, S.V., Huh, D.S., Venkataraman, A., 2011,

“Photo-Irradiated Biosynthesis of Silver Nanoparticles Using Edible

Mushroom Pleurotus florida and Their Antibacterial Activity Studies”,

Bioinorg. Chem Appl., 2011, pp. 1-7. doi:10.1155/2011/650979

[12] W. H. O., 1996, “Report of the WHO informal consultation on the evaluation

on the testing of insecticides”, CTD/WHO PES/IC/., 96(1), pp. 69

[13] Abbott, W.S., 1925, “A method of computing the effectiveness of an

insecticide”, J. Econ. Entomol., 18, pp. 265–266

[14] Upstone, S.L., 2000, “Ultraviolet/Visible Light Absorption Spectrophotometry

In Clinical Chemistry” John Wiley & Sons Ltd, Chichester

[15] Hemant, K.C., Narendra, V.B, Narayan, S.K., Dushyant, C.K, Ganesh, N.S.,

2012, “Synthesis and Characterization of Poly.meric Composites Embeded

with Silver Nanoparticles”, World J. Nano Sci. Engi., 2, pp.19-24.

doi.org/10.4236/wjnse.2012.21004

[16] Finney, D.J., 1971, “Probit Analysis”, 3rd ed. Cambridge University

Press,UK.

[17] Bar, H., Bhui, D.K., Sahoo, G.P., Sarkar, P., Pyne, S., Misra, A., 2009, “Green

synthesis of silver nanoparticles using seed extract of Jatrophacurcas”, Coll.

Surf. A-Physicochem Engineering Aspec., 348, pp. 212–216

[18] Das, R., Nath, S.S., Chakdar, D., Gope, G., and R. Bhattacharjee, 2009,

“Preparation of Silver Nanoparticles and Their Characterization”, J. nanotech.

Online, 5, pp. 1-6.

28 Anjali Rawani

[19] Nirmala, R., Faheem, A.S., Kanjwal, M.A., Lee, J.H., 2010, “Synthesis and

characterization of bovine femur bone hydroxyapatite containing silver

nanoparticles for the biomedical applications”, J. Nanoparticle Res., 10, pp.

9944–9950

[20] Raghunandan, D., Sarka, N., Choudhury, J., Roy, S.G., Bhattacharjee, A.,

2010, “Analysis of vibrational spectra of buckminsterfullerene (C60) using the

lie algebra”, Phys Sci. Technol., 2010; 6, pp. 60–63.

[21] Marimuthu, S., Rahuman, A.A., Rajakumar, G., Santhoshkumar, T., Kirthi,

A.V., 2011, “Evaluation of green synthesized silver nanoparticles against

parasites”, Parasitol. Res., 108, pp. 1541-1549.

[22] Priyadarshini, K.A., Murugan, K., Panneerselvam, C., Ponarulselvam, S.,

Hwang, J.S., Nicoletti, M., 2012, “Biolarvicidal and pupicidal potential of

silver nanoparticles synthesized using Euphorbia hirta against Anopheles

stephensi Liston (Diptera: Culicidae)”, Parasitol. Res., 111 (3), pp. 997-1006.

[23] Sareen, S., Pillai, J, Chandramohanakumar, RK., Balagopalan, M., 2012,

“Larvicidal Potential of Biologically Synthesised Silver nanoparticles against

Aedes Albopictus”, Res. J. Recent Sci., 1, pp. 52- 56

[24] Suman, T.Y., Elumali, D., Kaleena, P.K., Rajasree, S.R.R., 2013, “GCMS

analysis of bioactive components and synthesis of silver nanoparticle using

Ammannia baccifera aerial extract and its larvicidal activity against malaria

and filariasis vectors”, Ind. Crop. Prod., 47, pp. 39-245.

[25] Naik, B., Desai, V., Kowshik, M., Prasad, M.V., Fernando, G.F., Ghosh, N.N.,

2011, “Synthesis of Ag/AgCl–mesoporous silica nanocomposites using a

simple aqueous solution-based chemical method and a study of their

antibacterial activity on E. coli”, Particuology, 9, pp. 243–247. doi:

10.1016/j.partic.2010.12.001