The Journal of Basic & Applied Zoology (2016) 74, 37–40
H O S T E D BY
The Egyptian German Society for Zoology
The Journal of Basic & Applied Zoology www.egsz.org www.sciencedirect.com
Green synthesis of silver nanoparticles using Manihot esculenta leaves against Aedes aegypti and Culex quinquefasciatus K. Velayutham a, R. Ramanibai a,*, M. Umadevi b a b
Unit of Aquatic Biodiversity, Department of Zoology, University of Madras, Guindy Campus, Chennai 600025, Tamil Nadu, India Department of Physics, Mother Teresa Women’s University, Kodaikanal 624 101, Tamil Nadu, India
Received 21 April 2016; revised 25 May 2016; accepted 11 June 2016
KEYWORDS Silver nanoparticles; X-ray techniques; Electron microscopy; Mosquito larvae
Abstract To investigate the silver nanoparticles synthesis using leaves aqueous extract of Manihot esculenta against two important mosquito species, Aedes aegypti and Culex quinquefasciatus. The synthesized silver nanoparticles were monitored by UV–vis spectrophotometer and further characterized by XRD, FESEM and HRTEM. Aqueous extract of M. esculenta appeared to be effective against A. aegypti (LC50 = 66.14 mg/mL; LC90 = 172.41 mg/mL) and C. quinquefasciatus (LC50 = 61.60 mg/mL; LC90 = 184.80 mg/mL). The aqueous silver nitrate 1 Mm solution tested was third instar of A. aegypti and C. quinquefasciatus (LC50 = 76.96 mg/mL; LC90 = 230.88 mg/mL and LC50 = 84.06 mg/L; LC90 = 252.78 mg/mL). The 0.5 mM synthesized Ag PNs against A. aegypti (LC50 = 4.53 mg/mL; LC90 = 13.59 mg/mL); C. quinquefasciatus (LC50 = 3.46 mg/mL; LC90 = 10.38 mg/mL). The 1 mM silver solution synthesized Ag NPs tested were A. aegypti LC90 = 9.84 mg/mL) C. quinquefasciatus (LC50 = 3.21 mg/mL; (LC50 = 3.08 mg/mL; LC90 = 11.24 mg/mL). The control showed nil mortality in the concurrent assay. This is a perfect ecological and inexpensive approach for the control of mosquito larvae. Ó 2016 The Egyptian German Society for Zoology. 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/).
Introduction Mosquitoes are one of the best-known vectors of more than a few diseases such as zika, dengue fever and Japanese encephalitis. These diseases are endemic to mainly muggy countries causing millions of deaths. Larviciding is a doing well way of * Corresponding author. Fax: +044 22300899. E-mail addresses:
[email protected], biodiversityrr8@gmail. com (R. Ramanibai). Peer review under responsibility of The Egyptian German Society for Zoology.
sinking mosquito densities in their propagation places earlier than they appear into adults (Govindarajan et al., 2014). Insecticide applications, although highly efficacious against the target vector species control, are facing a threat due to the enlargement of resistance to chemical insecticides resulting in rebounding vectorial capability (Mehdi et al., 2012). Green synthesis of nano particle from plant extract may be a suitable alternative vector control approach in this regard (Makarov et al., 2014). Nanotechnology concerns with the development of tentative processes for the synthesis of nanoparticles of different sizes, shapes and controlled disparity (Ahmed et al., 2016). It
http://dx.doi.org/10.1016/j.jobaz.2016.06.002 2090-9896 Ó 2016 The Egyptian German Society for Zoology. 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/).
38 offers numerous benefits of eco-friendliness and compatibility for pharmaceutical and biomedical applications as they do not use deadly chemicals in the synthesis protocols (Bhosale et al., 2014). Manihot esculenta (Crantz) is an economic crop of India. Cassava which has been reported to have antimicrobial, antimalarial and leishmanicidal activities (Melo et al., 2009; Punthanara et al., 2009). The aim of this study was to investigate the silver nanoparticles synthesis using leaves aqueous extract of M. esculenta against two important mosquito species, A. aegypti and C. quinquefasciatus.
K. Velayutham et al. XRD analysis The XRD patterns of vacuum dried synthesized using aqueous leaf extract of M. esculenta. The 0.5 mM synthesized a number of Bragg reflections with 2h values of 17.50°, 28.84° and 35.44° sets of lattice planes were observed which may be indexed to the (002), (121) and (220) facts of silver. The 1 mM synthesized peaks at 38.02°, 34.69°, 44.39°, 64.62 and 77.40° correspond to the (1 0 1), (1 0 0), (2 0 4), (1 4 4) and (2 0 2), face centered cubic structures. FESEM analysis
M. esculenta leaves were collected from Karadimalai, Vellore district and an identity authenticity plant taxonomists.
FESEM micrograph shows the synthesized nanoparticles were (0.5 and 1 mM) spherical and aggregated shape. The EDX attachment with the FESEM was known to provide information on the chemical analysis of the fields that are being investigated or the composition at specific locations. In representative profile of the spot EDX analysis was obtained by focusing on Ag NPs.
Synthesis of Ag NPs
HR-TEM analysis
Materials and methods Collection of plant sample
Aqueous solutions of silver nitrate (AgNO3) (Sigma-Aldrich, Mexico) were prepared (0.5 mM and 1 mM) with distilled water. A known volume 20 mL of leaf extract was added, drop by drop, to 80 mL of 0.5 mM and 1 mM aqueous solution of silver, shaking continuously and gauged to 100 mL using distilled water, observing a brownish color, indicating the formation of Ag NPs. The synthesized AgNPs were characterized UV, XRD, FE-SEM and HR-TEM analysis. Rearing mosquito larvae Mosquito A. aegypti larvae were collected, maintained and reared in the laboratory for larvicidal bioassay for Kamaraj et al. (2008) method, minor modifications. Larvicidal bioassay One gram of aqueous extract was first dissolved in 100 mL of distilled water (stock solution). The larvicidal activity was assessed following WHO (1996) as per the method of Rahuman et al. (2000), minor modification. Data analysis
The biosynthesized Ag NPs were analyzed by HR-TEM to find the information of morphology size of nanoparticles. These micrographs show individual silver particles as well as a number of aggregated and spherical shapes. Under careful inspection of such images, these assemblies were found to be aggregates of silver nanoparticles. Larvicidal activity In the present study, the larvicidal aqueous crude leaf extracts, AgNO3 and synthesized Ag NPs of M. esculenta were prominent; however, the highest mortality was found in synthesized Ag NPs against third instar larvae of A. aegypti and C. quinquefasciatus at the concentration of 10 mg/L. The larvae mortality was observed aqueous leaf extract of M. esculenta (Fig. 1). The LC50 values of M. esculenta aqueous leaf extract appeared to be effective against A. aegypti (LC50 = 66.14 mg/mL; LC90 = 172.41 mg/mL) and C. quinquefasciatus (LC50 = 61.60 mg/mL; LC90 = 184.80 mg/mL) (Table 1). Most considerable mortality was evident after the treatment of silver nanoparticles. The aqueous AgNO3 1 Mm solution tested was third instar of A. aegypti and C. quinquefasciatus showed in (Fig. 2). The
The larvae mortality data were subjected to probit analysis by Reddy et al. (1992). Results Characterization of the synthesized nanoparticles UV–vis spectrum analysis Initially the aqueous extract was light green and upon providing the silver salt, it turned light brown color to dark brown color. The presence of nanoparticles was confirmed by obtaining a spectrum in UV-vis range of 200–800 nm. The 0.5 mM peak at 422 nm was obtained due to the SPR of silver nanoparticles. The highest values were reached with 1 mM AgNO3 the system became stable after 24 min and band of 426 nm.
Figure 1
Larvicidal activity of M. esculenta aqueous leaf extract.
Green synthesis of silver nanoparticles using Manihot esculenta leaves Table 1 larvae.
Effect of exposure of Manihot esculenta aqueous extract and synthesized silver nanoparticles on the mortality of mosquito
Species
Extract/Materials
Aedes aegypti
Aqueous extract Synthesized AgNPs
Culex quinquefasciatus
Figure 2 solution.
39
Aqueous extract Synthesized AgNPs
(0.5 mM) (1 mM) (0.5 mM) (1 mM)
LC50 mg/mL
LCL–UCL mg/mL
LC90 mg/mL
LCL–UCL mg/mL
r2
66.14 4.53 3.08 61.60 5.14 3.46
61.10–71.61 3.93–5.23 2.42–3.92 54.64–69.45 4.52–5.84 2.66–4.50
198.42 13.59 9.84 184.80 25.66 10.38
172–224.03 11.21–17.29 7.21–11.14 163.71–207.29 19.12–29.89 8.24–12.21
0.959 0.989 0.918 0.995 0.988 0.997
Graph showing larvicidal activity 1 mM AgNO3
A. aegypti (LC50 = 76.96 mg/mL; LC90 = 230.88 mg/mL) and C. quinquefasciatus (LC50 = 84.06 mg/L; LC90 = 252.78 mg/ mL), respectively. The larvae mortality was observed synthesized Ag NPs (0.5 mM AgNO3 solution) using aqueous leaf extract of M. esculenta (Fig. 3A). The LC50 = 4.53 mg/mL; LC90 = 13.59 mg/mL A. aegypti; LC50 = 3.46 mg/mL; LC90 = 10.38 mg/mL C. quinquefasciatus (Table 1). The 1 mM synthesized Ag NPs percent mortality shown in (Fig. 3B). The A. aegypti (LC50 = 3.08 mg/mL; LC90 = 9.84 mg/mL) and C. quinquefasciatus (LC50 = 3.21 mg/mL; LC90 = 11. 24 mg/mL). The control (distilled water) showed nil mortality in the concurrent assay. The complete mortality was observed for synthesized Ag NPs for A. aegypti and C. quinquefasciatus at concentration 10 mg/L (Table 1). Discussion In the present study, the mosquito larvicidal activity of aqueous leaf extracts, AgNO3 and morality variations synthesized Ag NPs of M. esculenta was noted. Muthukumaran et al. (2015) reported the synthesized Ag NPs using Chomelia asiatica against A. aegypti and C. quinquefasciatus at deliberation 40 mg/ml. The present study synthesized Ag NPs at concentration 10 mg/ml control for mosquito larvae. Logeswari et al. (2013) have been reported the silver nanoparticles exhibited arresting colors, starting light yellow to dark brown. Prasad and Elumalai (2011) reported that absorption spectra of silver nanoparticles twisted in the reaction media have absorbance peak at 430–440nm. The development of silver nanoparticles using aqueous leaf extract of M.
Figure 3 Larvicidal activity of synthesized Ag NPs (A) 0.5 mM and (B) 1 mM.
esculenta was viewed by the color change from light green to dark brown. Kathireswari et al. (2014) reported that the XRD patterns of vacuum dried Ag NPs synthesized using the leaf extract of Phyllanthus niruri peaks at 28.3, 32.4, 38.7, 46.0 assigned to the (2 2 0), (3 1 1), (1 1 1), (4 2 0) planes of a faced center cubic lattice of silver. Therefore XRD results also suggest that crystallization of the bioorganic phase occurs on the plane of the Ag NPs. In conclusion, synthesis of silver nanoparticles using M. esculenta against mosquito larvicidal activity was attempted. The substantial property of synthesized nanoparticle was characterized using related techniques. Further research on Ag NPs could bring a very promising target drug which can be used for protecting mosquito control.
40 Acknowledgements The authors are thankful to University Grants CommissionIndia, for providing financial assistance (RR) (F. No. 42630/2013 SR). References Ahmed, S., Ahmad, M., Swami, B.L., Ikram, S., 2016. A review on plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: a green expertise. J. Adv. Res. 7, 17–28. Bhosale, R.R., Kulkarni, A.S., Gilda, S.S., Aloorkar, N.H., Osmani, R.A., Harkare, B.R., 2014. Innovative eco-friendly approaches for green synthesis of silver nanoparticles. Int. J. Pharm. Sci. Nanotechol. 7 (1), 1. Govindarajan, M., Ramya, A., Sivakumar, R., 2014. Mosquito larvicidal properties of Mirabilis jalapa (Nyctaginaceae) against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). Indian J. Med. Res. 140 (3), 438–440. Kamaraj, C., Rahuman, A.A., Bagavan, A., 2008. Antifeedant and larvicidal effects of plant extracts against Spodoptera litura (F.) Aedes aegypti L. and Culex quinquefasciatus say. Parasitol. Res. 103, 325–331. Kathireswari, P., Gomathi, S., Saminathan, K., 2014. Plant leaf mediated synthesis of silver nanoparticles using Phyllanthus niruri and its antimicrobial activity against multi drug resistant human pathogens. Int. J. Curr. Microbiol. App. Sci. 3 (3), 960–968. Logeswari, P., Silambarasan, S., Abraham, J., 2013. Ecofriendly synthesis of silver nanoparticles from commercially available plant powders and their antibacterial properties. Sci. Iranica 20 (3), 1049–1054. Makarov, V.V., Love, A.J., Sinitsyna, O.V., Makarova, S.S., Yaminsky, I.V., Taliansky, M.E., Kalinina, N.O., 2014. Green nanotech-
K. Velayutham et al. nologies: synthesis of metal nanoparticles using plants. Acta Nat. 6 (1), 35–44. Mehdi, S.H., Qamar, A., Khan, I., Jacob, P., 2012. Larvicidal and IGR potential of Ocimum tenuiflorum and Datura alba leaf extracts against malaria vector. Eur. J. Exp. Biol. 2 (4), 1370–1375. Melo, F.M.P., Fiore, M.F., Moraes, L.A.B., Silva- tenico, M.E., Scramin, S., Teixeira, M.A., Melo, I.S., 2009. Antifungal compound produced by the cassava endophyte Bacillus pumilus MAIIIM4A. Sci. Agric. 66, 583–592. Muthukumaran, U., Govindarajan, M., Rajeswary, M., 2015. Mosquito larvicidal potential of silver nanoparticles synthesized using Chomelia asiatica (Rubiaceae) against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). Parasitol. Res. 114 (3), 989–999. Prasad, T.N.V.K., Elumalai, E.K., 2011. Biofabrication of Ag nanoparticles using Moringa oleifera leaf extract and their antimicrobial activity. Asian Pacific J. Trop. Biomed. 1 (6), 439–442. Punthanara, S., Chairatanayuth, P., Vijchulata, P., Surapat, S., Kuntho, U., Narongwanichakarn, W., 2009. Effects of cassava hay supplementation on antibacterial activity of the lactoperoxidase system in raw milk of dairy cows. Kasetsart J. Nat. Sci. 43, 486–496. Rahuman, A.A., Gopalakrishnan, G., Ghouse, B.S., Arumugam, S., Himalayan, B., 2000. Effect of Feronia limonia on mosquito larvae. Fitoterapia 71 (5), 553–555. Reddy, P.J., Krishna, D., Murthy, U.S., Jamil, K., 1992. A Microcomputer FORTRAN program for rapid determination of lethal concentration of biocides in mosquito control. CABIOS 8, 209– 213. World Health Organization, 1996. Instructions for Determining the Susceptibility or Resistance of Mosquito Larvae to Insecticides WHO/VBC/81.807. WHO, Geneva.