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Artificial Cells, Nanomedicine, and BiotechnologyAn International Journal

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Structural characterization and evaluationof mosquito-larvicidal property of silvernanoparticles synthesized from the seaweed,Turbinaria ornata (Turner) J. Agardh 1848

Paramasivam Deepak, Rajamani Sowmiya, Rajendiran Ramkumar,Govindasamy Balasubramani, Dilipkumar Aiswarya & Pachiappan Perumal

To cite this article: Paramasivam Deepak, Rajamani Sowmiya, Rajendiran Ramkumar,Govindasamy Balasubramani, Dilipkumar Aiswarya & Pachiappan Perumal (2016): Structuralcharacterization and evaluation of mosquito-larvicidal property of silver nanoparticlessynthesized from the seaweed, Turbinaria ornata (Turner) J. Agardh 1848, Artificial Cells,Nanomedicine, and Biotechnology

To link to this article: http://dx.doi.org/10.1080/21691401.2016.1198365

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ORIGINAL ARTICLE

Structural characterization and evaluation of mosquito-larvicidal property of silvernanoparticles synthesized from the seaweed, Turbinaria ornata (Turner)J. Agardh 1848

Paramasivam Deepaka, Rajamani Sowmiyaa, Rajendiran Ramkumarb, Govindasamy Balasubramania,Dilipkumar Aiswaryaa and Pachiappan Perumala

aDepartment of Biotechnology, School of Biosciences, Periyar University, Salem, Tamil Nadu, India; bDepartment of Biotechnology,Padmavani Arts & Science College for Women, Salem, Tamil Nadu, India

ABSTRACTThe silver nanoparticles synthesized from Turbinaria ornata (To-AgNPs) showed spherical with crystallinenature (20–32 nm) was evaluated against fourth instar larvae of three mosquitoes. The maximum activityof To-AgNPs was recorded on Aedes aegypti followed by Anopheles stephensi and Culex quinquefasciatuswith the following lethal concentration values (lg/ml): LC50 of 0.738, 1.134, and 1.494; and LC90 of 3.342,17.982, and 22.475, respectively. The obtained respective values (lg/ml) vis-a-vis aqueous extract (To-AE)were: 2.767 and 40.577; 4.347 and 158.399, and 7.351 and 278.994. The findings revealed that To-AgNPscould form a base for the development of an eco-friendly, low-cost pesticide.

ARTICLE HISTORYReceived 6 May 2016Revised 28 May 2016Accepted 28 May 2016Published online 20 June2016

KEYWORDSAedes aegypti; larvicidalactivity; silver nanoparticles;Turbinaria ornata

Introduction

Mosquitoes are blood sucking insects, which cause numerousdiseases to millions of people throughout the world (WHO2009). Through their bites, they transmit parasites and causeskin allergy and diseases to human beings. There are 350–500million clinical cases of malaria per year with about one mil-lion deaths. Especially in India, every year two million malariacases are being reported (Veerakumar et al. 2014). The mos-quito species, Aedes aegypti is a vector of chikungunya anddengue virus (Flavivirus) and it is an anthropophilic mosquito,which has got an intimate relationship with human by exhib-iting several behavioral traits like oviposition in man-madeand man-used natural and artificial containers (Yu et al. 2015).It causes chikungunya, a severe viral disease which has beenrecently considered to be an important public health problemin India and in other countries like Senegal and West Africa(Yamar et al. 2005). Dengue fever is prevalent throughout thetropics and subtropics. It is the most significant mosquitospread viral disease and a major international public healthconcern. Dengue hemorrhagic fever (DHF) is caused by den-gue virus which belongs to the genus, Flavivirus, of the familyFlaviviridae and it includes serotypes 1, 2, 3, and 4 (Den-1,Den-2, Den-3, and Den-4) (Veerakumar et al. 2013). The WorldHealth Organization estimates that around 2.5 billion peopleare at risk to dengue and such infections have dramaticallyincreased in recent decades due to increased urbanization,trade and travel (WHO 2010). As there is no effective drug orvaccine against dengue, the only way of its prevention is to

combat the disease-carrying mosquitoes from breeding andbiting humans.

In view of the effectiveness and recognized non-toxiceffects on non-target organisms, the plant-based insecticideshave been regularly used to control the mosquitoes(Govindarajan and Rajeswary 2015, Jayaraman et al. 2015,Santhosh et al. 2015, Suganya et al. 2013). Of late, mosquitoci-dal silver nanoparticles (AgNPs) are being synthesized fromthe oil-seed extracts and seaweed-borne compounds(Murugan et al. 2015, Pavela 2015). Nanoparticles play a majorrole in disease diagnostics, sensing, imaging, delivering geneand drug, artificial implants and in tissue engineering(Morones et al. 2005). Plant based NPs synthesis is advanta-geous over chemical and physical methods, as it is cheaper,rapid, single step method and eco-friendly, does not requirehigh pressure, energy, temperature, and the use of highlytoxic chemicals (Huang et al. 2007). Moreover, NPs synthesisusing different plant parts are more advantageous rather thanwith microbes because it eliminates the elaborate process ofmaintaining cell cultures (Shankar et al. 2004).

As it is essential to find a novel mosquito-insecticide, thereis growing interest in looking for natural alternatives espe-cially from the widely distributed but under exploited sea-weeds. Therefore, it is necessary to explore the bioactivecomponents of seaweeds so as to assess their mosquitocidalactivity. Understanding the seaweed chemical constituentswould provide information on the structure-activity relation-ships of active compounds that are responsible for the

CONTACT Pachiappan Perumal perumalarticles@gmail.com Department of Biotechnology, School of Biosciences, Periyar University, Salem 636 011,Tamil Nadu, India� 2016 Informa UK Limited, trading as Taylor & Francis Group

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insecticidal action (Dias and Moreas 2014, Yu et al. 2014).Recently, some seaweed-based AgNPs synthesis have beenreported; Padina tetrastromatica (Rajeshkumar et al. 2012),Sargassum wightii (Shanmugam et al. 2013), Codium capitatum(Kannan et al. 2013), Padina gymnospora (Singh et al. 2013),Chaetomorpha linum (Ragupathi Raja Kannan et al. 2013),Gracilaria corticata (Kumar et al. 2013), Hypnea musciformis(Ganapathy Selvam and Sivakumar 2014), Caulerpa scalpellifor-mis (Murugan et al. 2015) Caulerpa racemosa (Kathiraven et al.2015). The Turbinaria ornata extract have been reported tohave antibacterial (Vijayabaskar and Shiyamala 2011), anti-coagulant (Arivuselvan et al. 2011), anti-inflammatory (Ananthiet al. 2011), anti-oxidant properties (Kelman et al. 2012,Chakraborty et al. 2013). In view of the above, this study wasfocused on the structural characterization and mosquito-larvi-cidal potential of the AgNPs synthesized from the seaweed,Turbinaria ornata (Turner) J. Agardh.

Materials and methods

Collection of seaweed samples

Fresh marine brown algal samples were collected from theMandapam, Ramanathapuram District, Tamil Nadu, Southeastcoast of India (9� 22’ N; 78� 52’ E). The marine macroalga,Turbinaria ornata (Turner) J. Agardh 1848 (Sargassaceae)(Figure 1) have been identified based on standard keys,(Dinabandhu 2010) and the identification was confirmedby Dr. N. Kaliaperumal, Principal Scientist (Retd.), CentralMarine Fisheries Research Institute, Mandapam Camp,Ramanathapuram District, India. The reference specimenshave been kept in the Department of Biotechnology, PeriyarUniversity, Salem. Silver nitrate (AgNO3) was purchased fromHi-media Pvt. Ltd, India. All the chemicals used in this studywere of analytical grade with maximum purity.

Mosquito culture

The fourth instar mosquito larvae of Aedes aegypti, Anophelesstephensi, and Culex quinquefasciatus collected from SalemDistrict, Tamil Nadu, India, were kept in plastic trays

containing tap water and maintained in laboratory condition.Biscuits were served as larval food. All the experiments werecarried out at 28 ± 2�C, 70–80% relative humidity and photo-period of 12 h in light and dark conditions, respectively.

Preparation of Turbinaria ornata aqueous extract (To-AE)

The fresh seaweed sample was washed thoroughly with tapwater and then with distilled water to remove marine soildebris and associated biota. The washed seaweeds were driedunder shadow for three weeks. Then the dried seaweeds wereground using electric blender. Aqueous extract was preparedby mixing 10 g of dried seaweed powder in 100 ml of steriledouble distilled water with constant stirring on a magneticstirrer. Finally, the extract was filtered with Whatman no. 1 fil-ter paper and the extract was used within 1 h.

Synthesis of silver nanoparticles (AgNPs)

Twelve milliliters of To-AE was treated with 88 ml of 1 mMAgNO3 (16.96 mg in 100 ml Milli-Q water) solution in anErlenmeyer flask and incubated at room temperature toobserve the color change from brownish yellow to brown solu-tion that indicates the formation of AgNPs (Chitra et al. 2015).

Characterization of the synthesized To-AgNPs

The silver nanoparticle formation was observed periodicallyusing UV–vis spectrophotometer (Cyber Lab UV-100, Millbury,MA) in the range of 300–800 nm operated at a resolution of1 nm. The synthesized AgNPs were subjected to centrifugationat 10,000�g for 10 min and the pellet was dissolved in deion-ized water, filtered, and air dried. The centrifuged andparched material containing silver nanoparticles was used forX-ray diffraction (XRD) (Advance power X-ray diffractometerD8, Brucker, Karlsruhe, Germany) and Fourier transform infra-red spectroscopy (FTIR) analysis (Perkin-Elmer Spectrum 2000,Waltham, MA). In addition, the presence of silver ions in thesample was analyzed by energy dispersive X-ray spectroscopy(EDX). The size and morphology of the To-AgNPs were ana-lyzed by field emission-scanning electron microscope (FE-SEM;JEOL, Model JFC-1600, Zeiss, Germany). The measurement ofparticle size and zeta potential of To-AgNPs was carried outby particle size analyzer system (Zeta-sizer, MalvernInstruments Ltd, Worcestershire, UK).

Mosquito larvicidal bioassay

The larvicidal efficacy of To-AE and synthesized To-AgNPs weretested as per the guidelines of World Health Organization pro-cedure (WHO 1996), with some modifications. Different con-centrations (10, 5, 2.5, 1.25, and 0.625 ppm) of To-AE and To-AgNPs were used, respectively, to test larvicidal activity on A.aegypti, A. stephensi, and C. quinquefasciatus mosquito larvaeand a control without test samples. Batches of 25 healthy 4thinstar larvae were added into the glass beaker containing250 ml of tap water with desired concentration of To-AgNPsFigure 1. Marine macro-alga, Turbinaria ornata (Turner) J. Agardh 1848.

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and To-AE for each mosquito species. The number of dead lar-vae was counted after 24 h of exposure and their percentagemortality was calculated. Three replicates were performed andcontrol group (Tap water) was assigned to each mosquito spe-cies. Percentage mortality was calculated using Abbott’s for-mula (Abbott 1925), Mortality (%)¼ X – Y/*100; where, X:survival in the untreated control and Y: survival in thetreated sample (Gnanadesigan et al. 2011).

Statistical analysis

The average larval mortality data were subjected to ProbitAnalysis (Finney 1971) for calculating LC50, LC90, and otherstatistics at 95% confidence limit with upper and lower confi-dence limit, and Chi-square values were calculated using theSPSS 16.0 software (Armonk, NY). Results were considered tobe statistically significant at p< 0.05.

Results

Characterization of To-AgNPs

The formation of AgNPs was observed visually by change incolor of aqueous extract of T. ornata from brownish yellow tobrown with 1 mM AgNO3. The To-AE without AgNO3 did notshow any color change (Figure 2). The bio-reduction of Agþ ionshad occurred within 10 min, with a strong surface plasmon res-onance at 420 nm (Figure 3), thus indicating the formation ofAgNPs. The XRD analysis of To-AgNPs showed the 2h values of38.00�, 46.55�, 65.06�, and 77.40� corresponding to (1 1 1), (2 00), (2 2 0) and (3 1 1), respectively (Figure 4). The FTIR spectrumof To-AE (Figure 5(a)) showed bands at, 3415.72, 2070.591576.46 and 1399.15 cm�1. The band at 3415.72 cm�1 couldbe assigned to the O�H stretching vibration of alcoholgroups. The band at 2070.59 cm�1 corresponds to the N¼Cstretching vibration of miscellaneous groups. The bands at1576.46 and 1399.15 cm�1 were corresponding to the C–Ostretching vibration of carboxylic acids. FTIR spectrum of To-AgNPs (Figure 5(b)) showed the presence of bands at3402.60, 2201.21, 1572.62, 1401.87, 1481.88, 1260.35,1123.26, 1036.32, 924.47, 830.85, 762.70, 651.74, 697.18,

618.05, and 429.22 cm�1. The band at 3402.60 cm�1 couldbe assigned to the O–H stretching vibration of alcoholgroups. Similarly, the band at 2201.21 cm�1 signals to theC�C stretching vibration of alkynes groups. The bands at1572.62 and 1401.87 cm�1 also represents the C–O stretch-ing vibration of carboxylic acids. The band at 1481.88 andcm�1 consigned to the C–C stretching vibration of aromaticsgroups. The bands at 1260.35, 1123.26 and 1036.32 cm�1

were signaling to the C–H stretching vibration of alkyl hal-ides. The bands at 924.47 and 830.85 cm�1 were of miscel-laneous groups. The band at 762.70 cm�1 represents theC–Cl stretching vibration of alkyl halides. The bands at651.74 and 697.18 cm�1 were of alkynes with C�H stretch-ing vibrations. Finally, the bands at 618.05 and 429.22 cm�1

were assigned to the alkynes of C�H and alkyl halides ofC–H wag groups. In addition, the FTIR peaks indicated thepresence of alcohols, alkynes, aromatics, carboxylic acids,and alkyl halides. The FE-SEM micrograph of To-AgNPs(Figure 6(a) and (b)) clearly indicated the presence of spher-ical particles with size ranges between 20 and 32 nm. Theenergy dispersive X-ray spectroscopy (EDX) analysis ofTo-AgNPs confirmed the presence of elemental silver as themajor constituent (Figure 6(c)). The zeta potential analysis ofTo-AgNPs showed the negative value �2.86 mV, confirmingthat the nanoparticles are in colloidal state (Figure 7(a)). Theparticle size analysis exposed the Z-average of To-AgNPs are655.6 (d.nm) and PdI is 0.926 (Figure 7(b)).

Mosquito larvicidal activity of To-AE and To-AgNPs

The percentage mortality of To-AE and To-AgNPs for 4th instarlarvae of A. aegypti, A. stephensi, and C. quinquefasciatus aredepicted in Figures 8 and 9. The obtained LC50 and LC90 valuesare presented in Tables 1 and 2. The effective LC50 and LC90 val-ues (lg/ml) of To-AE were: 2.767 and 40.577 (A. aegypti) fol-lowed by 4.347 and 158.399 (A. stephensi), and 7.351 and278.994 (C. quinquefasciatus), respectively. Whereas the To-AgNPs showed the LC50 and LC90 values (lg/ml) of: 0.738 and3.342 (A. aegypti), 1.134 and 17.982 (A. stephensi), and 1.494 and22.475 (C. quinquefasciatus), respectively. The control did notshow any mortality. v2 value was significant at p< 0.05 level.

Figure 2. Synthesis of silver nanoparticles using T. ornata: (a) AgNO3.(b) Aqueous extract of T. ornata. (c) T. ornata- silver nanoparticles (To-AgNPs). Figure 3. UV–Vis absorption spectra of To-AgNPs.

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Discussion

Nano-biotechnology is fast-emerging to be an important dis-cipline of modern science and engineering, which providesscope for the synthesis of raw materials of distinctive sizes,shapes, and compositions. The nanoparticles of silver, gold,and palladium are being widely used in the development ofmedical and pharmaceutical products (Song et al. 2010). Theearlier study has reported that nano or colloidal state ofmetallic silver gives different color to aqueous solution due tothe excitation of electrons from the valence bond to conduc-tion bond (Mulvaney 1996). Currently, the formation of AgNPswas confirmed by the color change of seaweed aqueousextract, when AgNO3 was added. The occurrence of browncolor was due to the excitation of ions that signals a strong

Figure 4. XRD pattern of synthesized To-AgNPs.

Figure 5. FTIR spectrum: (a) To-AE. (b) synthesized To-AgNPs.

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surface plasmon vibration at 420 nm, which indicated the for-mation of silver nanoparticles. The XRD analysis of To-AgNPsshowed four intense peaks (1 1 1), (2 0 0), (2 2 0), and (3 1 1),that are in agreement to the Bragg’s reflection of silver nano-crystals (Lu et al. 2003). Further, the XRD-pattern with sharpBragg peaks are in agreement with the Joint Committee onPower Diffraction Standards (JCPDS no. 040783), confirmingthe crystalline nature of the AgNPs, as reported earlier(Gnanadesigan et al. 2012). The observed other minorunassigned peaks in their vicinity might have been due to thecapping agent that stabilizes the nanoparticles. Independentcrystallization of the capping agents, is ruled out due to theprocess of centrifugation and redispersion of the pellet indeionized distilled water after nanoparticle formation as partof purification process (Haldar et al. 2013). The presence offunctional groups viz: alkane, methylene, alkene, amine, andcarboxylic acids in the aqueous plant leaf extract, has alreadybeen proved to be of potential reducing agents for the syn-thesis of AgNPs (Cho et al. 2005). Interestingly, the FTIR spec-trum of To-AE (Figure 5(a)) showed the presence of aminegroups and carboxylic acids, which might act as a cappingagent, for the reduction of silver ions. The recent study hasreported that the different functional groups present in theaqueous extract would stabilize the AgNPs (Chanthini et al.2015). The FE-SEM micrograph of To-AgNPs showed spherical

shape, which evidenced the nature of silver that have alreadybeen reported by previous researchers (Merin et al. 2010,Veerakumar et al. 2013). The metallic silver nanocrystals thatgenerally show typical absorption peak approximately at 3 keVdue to surface plasmon resonance (Haldar et al. 2013). TheEDX pattern of To-AgNPs showed strong signals for Ag,whereas the Cl and O peaks might be originated from theaqueous extract. In addition, the zeta potential measurementof To-AgNPs showed a sharp peak at �2.86 mV suggestingthat the surface of the nanoparticles is negatively charged.The zeta potential value of To-AgNPs is an indication of repul-sive force which could be used to predict the long term sta-bility of the product (Patil et al. 2012).

Several researchers have found that the seaweeds possessgood insecticidal property like that of the terrestrial plants-derived bio-insecticides (Ghosh et al. 2012, Oliveira et al.2010, Yu et al. 2014). Presently, we have investigated the larvi-cidal effect of T. ornata on the fourth instar larvae of Aedesaegypti, Anopheles stephensi, and Culex quinquefasciatus.Generally, the fourth instar larvae are considered to be havingmore immune-competency than the younger larvae (Patilet al. 2012, Vogelweith et al. 2013). To-AE showed moderatelarvicidal effect with the lethal concentrations (LC) at the LC50

values of (lg/ml): 2.767, 4.347, and 7.351 and LC90 values of(lg/ml): 40.577, 158.399, and 278.994 against A. aegypti,

Figure 6. FE-SEM images of To-AgNPs. (a) magnified at 50,000�. (b) magnified at 100,000�. (c) EDX spectrum of To-AgNPs.

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A. stephensi, and C. quinquefasciatus, respectively. Therecorded inhibitory concentrations of To-AgNPs (lg/ml)towards the highest mortality were: LC50; 0.738, 1.134, and1.494; and LC90; 3.342, 17.982, and 22.475 against A. aegypti,A. stephensi, and C. quinquefasciatus, respectively. These mor-tality effects could have been due to the presence of various

phytal-constituents of seaweed, T. ornata. The phytal-constitu-ents, including phenols, terpenoid, flavonoids, saponin, andalkaloids of the seaweed, Hypnea musciformis might beresponsible for the larvicidal property (Roni et al. 2015).

Figure 7. (a) Zeta potential values of To-AgNPs. (b) Particle size distribution of To-AgNPs.

0.625 1.25 2.5 5 100

20

40

60

80

100 A. aegypti

A. stephensi

C. quinquefasciatus

Concentration of To-AE in µg/ml

% o

f m

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Figure 8. Percentage mortality of A. aegypti, A. stephensi, and C. quinquefasciatususing To-AE.

0.625 1.25 2.5 5 100

20

40

60

80

100

A. aegypti

A. stephensi

C. quinquefasciatus

Concentration of To-AgNPs in µg/ml

% o

f m

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Figure 9. Percentage mortality of A. aegypti, A. stephensi, and C. quinquefasciatususing To-AgNPs.

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Recent study reported that, the bio-toxicity of Ulva lactuca-mediated AgNPs showed the LC50 and LC90 values (lg/ml):2.111 and 8.350; 3.090 and 10.292; 4.629 and 13.254; 5.261and 14.851 against the malarial mosquito larvae A. stephensi(1st to 4th instars), respectively (Murugan et al. 2015).

Some earlier researchers have found that the acetoneextracts of the marine seaweeds; Caulerpa scapelliformis,Dictyota dichotoma, Enteromorpha clathrata, E. intestinalis,and U. lactuca showed good activity against 4th instar lar-vae of A. aegypti with the following LC50 values (lg/ml):53.70, 61.65, 85.11, 67.70, and 91.20, respectively (Thangamand Kathiresan 1991). The larvicidal activity of acetoneextracts of seaweed, C. scalpelliformis against 2nd and 3rdinstar larvae of Culex pipiens with the following LC50 andLC90 values (lg/ml): 338.91 and 1891.31, respectively (Cetinet al. 2010). Recent report also confirms the larvicidal effi-cacy of seaweed; Caulerpa scalpelliformis mediated AgNPsthat showed maximum LC50 and LC90 values (lg/ml) of 5.86and 12.38, respectively, for the 4th instar mosquito larvae of

C. quinquefasciatus (Murugan et al. 2015). The larvicidal activ-ity of seaweed, Hypnea musciformis-mediated AgNPs onA. aegypti (1st to 4th instars) showed the following LC50 andLC90 values (lg/ml): 18.14 and 246.59, 20.54 and 269.05,26.61 and 301.66, 27.99 and 319.30, respectively (Roni et al.2015).

Many reports have appeared on the larvicidal property ofterrestrial plants based AgNPs. The larvicidal activity of AgNPsfrom aqueous leaf extract of Sida acuta against the third-instarlarvae of C. quinquefasciatus (LC50: 26.13 and 130.30 lg/ml),A. stephensi (LC50: 21.92 and 109.94 lg/ml), and A. aegyptiLC50 (23.96 and 119.32 lg/ml), respectively (Veerakumar et al.2013). Recently, larvicidal activity of AgNPs and aqueous leafextract of Annona muricata were tested against A. aegypti(LC50: 12.58 and 51.13 lg/ml; LC90: 26.46 and 82.08 lg/ml),A. stephensi (LC50: 15.28 and 61.38 lg/ml; LC90: 31.91 and156.55 lg/ml) and C. quinquefasciatus (LC50: 18.77 and88.72 lg/ml; LC90: 35.72 and 199.67 lg/ml), respectively(Santhosh et al. 2015).

Table 1. Larvicidal activity of T. ornata aqueous extract against fourth instar larvae of A. aegypti, A. stephensi, and C. quinquefasciatus at 24-hobservation.

Mosquitoes Concentration of To-AE 24-h mortality (%) ± SD LC50 lg/mL (LCL–UCL) LC90 lg/mL (LCL–UCL) v2

Control 0.0 ± 0.0

A. aegypti 0.62 26.66 ± 1.52 2.767 (2.087–3.728) 40.577 (20.760–132.244) 4.484*1.25 37.33 ± 0.572.50 48.00 ± 1.005.00 60.00 ± 1.73

10.00 73.00 ± 2.08

A. stephensi 0.62 21.33 ± 1.15 4.347 (3.004–7.449) 158.399 (49.115–2040.713) 5.480*1.25 36.00 ± 1.002.50 41.00 ± 0.575.00 57.33 ± 1.52

10.00 60.33 ± 1.00

C. quinquefasciatus 0.62 18.66 ± 1.52 7.351 (4.760–16.240) 278.994 (73.000–5718.211) 3.571*1.25 26.66 ± 0.572.50 37.33 ± 0.575.00 42.66 ± 2.08

10.00 54.66 ± 1.15

SD: standard deviation; LCL: lower confidence limit; UCL: upper confidence limit; v2: chi square test; *p<0.05, level of significance, values aremean ± SD of three replicates.

Table 2. Larvicidal activity of T. ornata AgNPs against fourth instar larvae of A. aegypti, A. stephensi, and C. quinquefasciatus at 24-hobservation.

Mosquitoes Concentration of To-AgNPs 24-h mortality (%) ± SD LC50 lg/mL (LCL–UCL) LC90 lg/mL (LCL–UCL) v2

Control 0.0 ± 0.0

A. aegypti 0.62 37.33 ± 1.52 0.738 (0.543–0.921) 3.342 (2.641–4.680) 13.530*1.25 62.66 ± 0.572.50 78.66 ± 3.055.00 97.33 ± 0.57

10.00 100.00 ± 0.0

A. stephensi 0.62 34.66 ± 0.57 1.134 (0.724–1.547) 17.982 (10.325–48.436) 4.60*1.25 50.66 ± 1.522.50 60.00 ± 1.005.00 73.33 ± 0.57

10.00 88.00 ± 1.00

C. quinquefasciatus 0.62 32.00 ± 1.00 1.494 (1.034–1.987) 22.475 (12.629–61.917) 5.343*1.25 48.00 ± 1.002.50 54.66 ± 1.525.00 74.66 ± 2.08

10.00 81.13 ± 1.52

SD: standard deviation; LCL: lower confidence limit; UCL: upper confidence limit; v2: chi square test; *p< 0.05, level of significance, values aremean ± SD of three replicates.

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Conclusion

In this study, we reported the mosquitocidal property of T.ornata-mediated AgNPs that was found to be higher whencompared to the larvicidal properties recorded by the previ-ous researchers. The nano-size and spherical shape of the syn-thesized AgNPs from T. ornata were confirmed through FE-SEM micrograph analysis and the To-AgNPs showed highmosquito-larvicidal potential.

Acknowledgements

The authors are grateful to the authorities of Periyar University for provid-ing the necessary facilities to carry out this work.

Disclosure statement

No potential conflict of interest was reported by the authors.

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