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. 2020 Jul 20;11:e00166. doi: 10.1016/j.parepi.2020.e00166

Larvicidal and antibacterial activity of aqueous leaf extract of Peepal (Ficus religiosa) synthesized nanoparticles

Namita Soni 1,⁎, Ramesh C Dhiman 1
PMCID: PMC7452144  PMID: 32885057

Abstract

In this study, the zinc oxide nanoparticles (ZnO NPs) and titanium dioxide nanoparticles (TiO2 NPs) were synthesized using the aqueous leaf extract of Ficus religiosa (Peepal tree). The synthesized nanoparticles were tested as larvicides against the larvae of Anopheles stephensi. Further, the synthesized nanoparticles were tested as antibacterial agents against the Escherichia coli (gram negative) and Staphylococcus aureus (gram positive) bacteria. The synthesized nanoparticles were characterized with UV-visible spectroscopy, X-rays powder diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). The larvicidal mortality was observed after 24 h and 48 h by probit analysis. The antibacterial activity was evaluated using the well diffusion method. The synthesized nanoparticles were irregular shape and varied size. The larvae of An. stephensi were found highly susceptible against the ZnO NPs than the TiO2 NPs and aqueous leaves extract. The highest mortality was observed in synthesized ZnO NPs against first to third instars of (LC50 50, 75, and 5 ppm) and 100% mortality in fourth instars of An. stephensi. The higher zone of inhibition was occurred against the E. coli. This report of present investigation revealed that the rapid biological synthesis of ZnO NPs and TiO2 NPs using aqueous leaf extract of F. religiosa would be effective potential larvicides for mosquito control as well as antimicrobial agents with eco-friendly approach

Keywords: Ficus religiosa, ZnO NPs, TiO2 NPs, Larvicides, Anopheles stephensi, Antimicrobial

1. Introduction

F. religiosa is commonly known as Peepal tree belonging to the family moraceae. It is also known as bodhi tree, pippala tree, peepul tree, pepal tree and ashwattha tree. It is a medicinal plant which is emerged as a good source of traditional medicine for the treatment of asthma, diabetes, diarrhea, epilepsy, gastric problems, inflammatory disorders, infectious and sexual disorders (Singh et al., 2011a). The leaves of F. religiosa contain tannic acid, leucine, isoleucine, methionine, tryptophan, threonine, glycine asparatic acid, serine and arginine, bark comprises of bergaptol and bergapten, the fruits consists of tyrosine and asparagines and seeds contain threonine, alanine and valine (Gupta and Singh, 2012), which are the good source of secondary metabolites used as larvicides and antimicrobial agents.

The larvicidal activity of crude hexane, ethyl acetate, petroleum ether, acetone, and methanol extracts of the leaf and bark of Ficus racemosa (Moraceae) has been assayed for their toxicity against the early fourth-instar larvae of Culex quinquefasciatus (Abdul Rahuman et al., 2008). The larvicidal efficacy of different extracts of F. benghalensis against Cx. quinquefasciatus, Ae. aegypti and An. stephensi has been investigated (Govindarajan, 2010). Further, larvicidal efficacy of different solvent leaf extracts of F. benghalensis against Cx. tritaeniorhynchus and An. subpictus has been determined (Govindarajan et al., 2011). Thereafter, the Larvicidal activity of Indian medicinal plants, Commiphora berryi, Commiphora pentandra, Pelargonium graveolens, Thevetia peruviana, Sesamum indicum, Ficus microcarpa, Melia dubia, C. bonplandianus, F. religiosa and Croton lacryma has been tested against Ae. aegypti mosquito (Deepa et al., 2015). The antimicrobial properties of Ficus extract have been reported (Mandal et al., 2000; Ao et al., 2008; Oyeleke et al., 2008; Annan and Houghton, 2008; Kuete et al., 2008; Kuete et al., 2009; Usman et al., 2009; Alimuddin et al., 2010; Lazreg Aref et al., 2010). Unfortunately, the secondary metabolites of plants have the slow reaction against the mosquitoes. Therefore, it is needed to develop the eco-friendly and rapid technology for the mosquito control as well as antimicrobial agent, so that people can be protected from the bacterial and vector borne diseases.

Recently, the use of metal nanoparticles is a great attention in this regards. The plants synthesized ZnO NPs and TiO2 NPs are the good, rapid and eco-friendly sources for mosquito control and antibacterial agents also. The synthesis and characterization of ZnO NPs (Singh et al., 2011b; Awwad et al., 2014; Shekhawat et al., 2014; Gnanasangeetha and Thambavani, 2014; Raju Kooluru and Sharada, 2014; Suganya et al., 2015; Noorjahan et al., 2015; Varghese and George, 2015; Manokari and Shekhawat, 2015; Manokari et al., 2016a, Manokari et al., 2016b; Fatimah et al., 2016; Sharmila Devi and Dhinesh, 2016; Pinto and Nazareth, 2016) and TiO2 NPs (Sundrarajan and Gowri, 2011; Rajakumar et al., 2012; Ganapathi Rao et al., 2015; Khadar et al., 2015; Valli and Geetha, 2015; Chatterjee et al., 2016; Mythreyi et al., 2016; Dobrucka, 2017; Patidar and Jain, 2017) using the plant extract has been reported.

Acaricidal, pediculocidal and larvicidal activity of ZnO NPs using wet chemical method has been reported (Kirthi et al., 2011). The antimicrobial activity of synthesized ZnO NPs using plants has been reported (Jeeva Lakshmi et al., 2012; Aswathi Sreenivasan et al., 2012; Divya et al., 2013; Mishra and Sharma, 2015; Raj et al., 2015; Narendhran and Sivaraj, 2016; Salih and Smail, 2016; Jeba Jane Ratney and Begila David, 2016; Paul et al., 2016).

The larvicidal activity of plant synthesized TiO2 NPs has been assessed (Rajakumar et al., 2015; Suman et al., 2015; Gandhi et al., 2016). The antimicrobial activity of synthesized TiO2 NPs using plants has been studied (Maurya et al., 2012; Malarkodi et al., 2013; Santhoshkumar et al., 2014; Murphin Kumar et al., 2014; Hariharan et al., 2017). Till now no review is available on the larvicidal and antibacterial activities of ZnO NPs and TiO2 NPs synthesized using aqueous leaf extract of Peepal (F. religiosa). In the present investigation, the ZnO NPs and TiO2 NPs were synthesized using the aqueous leaf extract of F. religiosa and access their larvicidal and antibacterial properties. This ZnO NPs and TiO2 NPs technology could be a rapid, green and, eco-friendly approach for mosquito control and used as antimicrobial also.

2. Materials and methods

2.1. Collection and leaf extract preparation

The fresh and green leaves of F. religiosa were collected from the nearest area of ICMR-National Institute of Malaria Research, India. The leaves were rinsed with tap water and then with distilled water to remove dust and other particles. The rinsed leaves were then air dried for 1–2 h. After then, approximately 20 g of leaves were cut into fine pieces and put into a 250 ml conical flask which containing 100 ml of distilled water. Boil the flask for 1 h at 50 °C on a magnetic stirrer. After 1 h, cooled the extract and filtered through the whatman-1 filter paper and store the leaves filtrate for the experiment.

2.2. Synthesis of ZnO and TiO2 NPs

The ZnO NPs were biosynthesized by co-precipitation method described by Singh et al. (2011a) with some modification. 20 ml of leaf extract was heated at 60 °C for 10 min on a magnetic stirrer. After then, 50 ml of 0.1 M of Zinc nitrate solution and 50 ml of 0.2 M sodium hydroxide solution were added drop wise under stirring. The mixture was continued stirred for 1 h on magnetic stirrer which resulting cream colored precipitate of zinc hydroxide formed. Then, the precipitate was collected by centrifugation at 4000 rpm for 15 min and washed with deionized water and ethanol. The ZnO NPs were collected after dried in hot air oven for 48 h at 45 °C.

The TiO2 NPs were biosynthesized by the following method described by Suman et al. (2015) with some modification. The aqueous solution of TiO(OH)2 (5 mmol/L) was prepared and used for synthesis of TiO2 NPs. 20 ml leaf extract of F. religiosa was boiled at 50 °C. Then 80 ml of aqueous solution of 5 mmol/L TiO(OH)2 were added in the leaf extract and boil for 4 h with continuous stirring. After then reduction mixture was centrifuged at 4000 rpm for 15 min and resulting pellet was collected. The TiO2 NPs were dried in hot air oven for 48 h at 45 °C.

2.3. Characterization of ZnO and TiO2 NPs

Optical properties of synthesized ZnO and TiO2 NPs were confirmed by UV–visible double beam spectroscopy (HALO DB-20) in 300–500 nm wavelength range. The XRD pattern of synthesized ZnO and TiO2 NPs were carried out using X-ray diffractometer (Bruker X-ray diffractometer D-8 Advance) Cu-Kα radiations (λ = 0.15406 nm) in 2θ range from 20° to 80°. The average size and shape of synthesized ZnO and TiO2 NPs were obtained by transmission electron microscopy (Tecnai G2). The morphology of synthesized ZnO and TiO2 NPs were examined by scanning electron microscope (model no. Zeiss EVO MA 10). The synthesized ZnO and TiO2 NPs were analyzed for elemental analysis by energy dispersive X-ray spectroscopy (Oxford Inca Energy 250).

2.4. Rearing of larvae

The larvae of An. stephensi were reared in deionized water containing glucose and yeast powder. The colony of An. stephensi was maintained in the laboratory at 27 °C with relative humidity of (75 ± 5%) and 14 h of photoperiod using the standard method with some modifications (Geberg et al., 1994).

2.5. Bioassay, data management and statistical analysis

ZnO NPs and TiO2 NPs, synthesized using aqueous leaves extract of F. religiosa were tested for their killing activities against the An. stephensi larvae (I-IV instar). The bioassay was assessed using the standard method (World Health Organization, 2005). An. stephensi larvae were placed in a container in micro-free deionized water. After that, ZnO and TiO2 NPs with different test concentrations in 100 mL deionized water were prepared in 250 mL beakers. Bioassays were conducted separately at five different concentrations using serial dilution method, of synthesized ZnO and TiO2 NPs (25, 50, 100, 150 and 250 ppm). To test the larvicidal activity of ZnO and TiO2 NPs, 20 larvae were separately exposed to 100 mL of test concentration. Similarly, the control (without ZnO and TiO2 NPs) was run to test the natural mortality. The experiments were replicated thrice to validate the results. Thereafter, we examined their mortality after 24 h and 48 h. The data on the efficacy was subjected to probit analysis (Finney, 1971). The control mortality was corrected by Abbott's formula (Abbott, 1925).

2.6. Antibacterial activity of ZnO and TiO2 NPs

The antibacterial activity of synthesized ZnO and TiO2 NPs was evaluated against E. coli and S. aureus. The antibacterial activity was determined using the well diffusion method. The wells were prepared on plates with Muller-Hinton agar (MHA) medium. Then, the plates were seeded with different bacterial strains using sterile swab. Four wells were prepared using gel puncture in each plate. Each well was loaded with 50 μL of different concentration of ZnO and TiO2 NPs (50, 150, 250 and 500 ppm). Then, the plates were incubated at 35 °C for 24 h and zone of inhibition was observed.

3. Results and discussion

3.1. Proposed mechanism of synthesis of ZnO and TiO2NPs through F. religiosa

Based on the experimental work that has been done, there are series of chemical reaction that takes place. The complete hydrolysis of zinc nitrate and dihydroxy(oxo)titanium with the aid of F. religiosa aqueous leaves extract solution should result in the formation of ZnO and TiO2 nanoparticles. The richly available carbohydrates, tannin, alkaloids, steroids, terpenoids, saponin, reducing sugar and favonoids in the plant extract acted as stabilizing and capping agents, respectively. Hence, the proposed principle of formation of ZnO and TiO2 NPs involves the ionization of zinc nitrate and hydroxylation of dihydroxy(oxo)titanium in an aqueous medium to give Zn2+ which was reduced by phytochemical principle present in the aqueous extract of F. religiosa, to generate ZnO, which further aggregates to ZnO and TiO2 NPs as shown in Eq. 1 and Eq. 2.

Unlabelled Image

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TiO(OH)2 + Ficus religiosa leaves extract∆Stirring TiO2 + H2O↑.

3.2. UV-visible analysis

The formation of ZnO and TiO2 NPs during the synthesis can be observed visually. Fig. 1a is the UV–vis absorption spectrum of ZnO NPs dispersed in deionized water and the figure shows the absorption peak at 358 nm. Shah et al. (2015) stated that the UV absorption spectrum for synthesized ZnO NPs was recorded at 330 nm. Similar results were observed by Yedurkar et al. (2016). Fig. 1b shows the UV-vis absorption spectrum of TiO2 NPs with an absorption peak at 450 nm. Valli and Geetha (2015) observed the UV absorption spectrum for synthesized TiO2 NPs at 447.3 nm.

Fig. 1.

Fig. 1

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UV-visible spectrum of aqueous leaves extract synthesized (a) ZnO NPs and (b) TiO2 NPs.

3.3. XRD analysis

The structure of ZnO and TiO2 NPs were determined in this study using a powder diffraction system with Cu-Ka x-ray tube (λ = 1.541836 A) was used. Fig. 2 depicts the XRD pattern of synthesized ZnO NPs scanned at 2θ range from 0 to 80 degree. Diffraction peaks at 31.66°, 34.34°, 36.15°, 47.45°, 56.46°, 62.72°, 67.86°, 68.97°, 76.79° can be assigned to (110), (002), (101), (102), (110), (103), (112), (201) and (202) plane. The strong and narrow peak denotes that the product has well crystalline nature of particles. Narendhran and Sivakumar, (2016) recorded the X-ray diffraction of ZnO NPs synthesized using the L. aculeate. The peaks at 2θ values of 31.79°, 34.42°, 36.26°, 47.59°, 62.80°, 65.84°, 67.96°, 68.30°, 72.12° and 76.53° correspondence to the crystal planes of (100), (002), (101), (102), (110), (103), (200), (112), (201), (004) and (202) of ZnO NPs. Similar results were reported by Vanathi et al. (2014) in which particles were synthesized using E. crassipes leaf extract.

Fig. 2.

Fig. 2

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XRD pattern of synthesized ZnO NPs.

Fig. 3 shows the XRD pattern of synthesized TiO2 NPs scanned at 2θ range from 0 to 80 degree. Diffraction peaks at 25.28°, 36.91°, 53.85°, 55.03°, 62.6°, 68.70° and 75.1° can be assigned to (110), (101), (211), (220), (204), (112) and (215) plane. Similar results were reported by Khadar et al. (2015).

Fig. 3.

Fig. 3

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XRD pattern of synthesized TiO2 NPs.

3.4. TEM analysis

The shape and size of synthesized ZnO and TiO2 NPs were obtained using the TEM. Fig. 4a shows the TEM images of synthesized ZnO NPs, which depict the irregular shape and varied size nanoparticles. Fig. 4b depicts the TEM images of TiO2 NPs, which were spherical in shape and size from 70.29 to 84.93 nm with calculated size of 77.61 nm and polydisperse. Zahir et al. (2014) recorded the TEM images of the synthesized Ag NPs and TiO2 NPs spherical, quite polydisperse and individual particles showed an average size of 12.82 ± 2.50 and 83.22 ± 1.50 nm, respectively. Similar results were obtained by (Rajakumar et al., 2012) and previous work (Table 1).

Fig. 4.

Fig. 4

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TEM images of synthesized (a) ZnO NPs and (b) TiO2 NPs.

Table 1.

Size Comparison of ZnO and TiO2 nanoparticles synthesized using the plants.

Plant species Plant part used Synthesized NPs Size (nm) References
Azadirachta indica, Emblica Officinalis leaf ZnO 51, 16 Gnanasangeetha and Thambavani, 2014
Pyrus pyrifolia leaf ZnO 45 Parthiban and Sundaramurthy, 2015
Ixora coccinea leaf ZnO 145.1 Yedurkar et al., 2016
Passiflora caerulea Leaf ZnO 200 Santhoshkumar et al., 2017
Bauhinia tomentosa Leaf ZnO 22–94 Sharmila et al., 2018
Pandanus odorifer leaf ZnO 90 Hussain et al., 2019
Allium sativum Skin ZnO 7.77 Modi and Fulekar, 2020
Psidium guajava Leaf TiO2 32.58 Santhoshkumar et al., 2014
Cynodon dactylon Leaf TiO2 13–34 Hariharan et al., 2017
Glycosmis cochinchinensis Leaf TiO2 45 Rosi and Kalyanasundaram, 2018
Cassia fistula Leaf TiO2 200 Swathi et al., 2019
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3.5. SEM-EDX analysis

The size and distribution of synthesized ZnO and TiO2 NPs were also confirmed by SEM shown in Fig. 5a-b. From the result it is evident that the morphology of ZnO NPs was irregular and TiO2 NPs was spherical in shape and polydisperse in nature, which is very similar to previous studies (Zahir et al., 2014; Rajakumar et al., 2012).

Fig. 5.

Fig. 5

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SEM images of synthesized (a) ZnO NPs and (b) TiO2 NPs.

Fig. 6 shows the EDX analysis of ZnO NPs 72.57% zinc and 27.42% of oxygen which confirm the elemental composition of ZnO NPs. Narendhran and Sivaraj (2016) showed the EDX analysis of ZnO nanoparticles 37.22% of zinc and 62.78% of oxygen which confirms the elemental composition of ZnO nanoparticles. The strong signals from the zinc atoms in the nanoparticles recorded and other signals from C and O atoms were observed using EDX analysis in Parthenium-mediated ZnO nanoparticles (Rajiv et al., 2013). The EDX analysis display the optical absorption peaks of ZnO nanoparticles and these absorption peaks were due to the surface plasmon resonance of ZnO nanoparticles (Ankanna and Savithramma, 2011).

Fig. 6.

Fig. 6

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EDX images of synthesized ZnO NPs.

Fig. 7 depicts the EDX analysis of TiO2 NPs 71.99% titanium and 28.01% of oxygen which confirm the elemental composition of TiO2 NPs. Santhoshkumar et al. (2014) showed the energy dispersive X-ray analysis study (EDX) which proves that the particles are crystalline in nature and indeed metallic TiO2 NPs. The similar results were reported in the previous studies by (Suman et al., 2015).

Fig. 7.

Fig. 7

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EDX images of synthesized TiO2 NPs.

3.6. Larvicidal activity of ZnO and TiO2 NPs

Larvicidal activity of F. religiosa leaf extract, synthesized ZnO and TiO2 NPs were evaluated against the larvae (I-IV) of An. stephensi at different concentrations (25, 50, 100, 150 and 250 ppm).

The larvae of An. stephensi were found highly susceptible to the ZnO NPs. The fourth instar larvae have shown the 100% mortality after 24 h of exposure. Whereas, the first instar (LC50 50 ppm), second instar (LC50 75 ppm) and third instar (LC50 5 ppm) larvae were observed with their probit equations and 95% confidential limit, R2, chi-square and p value after 24 h (Table 2). No mortality was observed in control group. The anti-parasitic activities have been assessed to determine the efficacies of synthesized zinc oxide nanoparticles (ZnO NPs) prepared by wet chemical method using zinc nitrate and sodium hydroxide as precursors and soluble starch as stabilizing agent against the larvae of cattle tick Rhipicephalus (Boophilus) microplus, Canestrini (Acari: Ixodidae); head louse Pediculus humanus capitis, De Geer (Phthiraptera: Pediculidae); larvae of malaria vector, An. subpictus, Grassi; and filariasis vector, Cx. quinquefasciatus, Say (Diptera: Culicidae) (Kirthi et al., 2011). The maximum efficacy was observed in zinc oxide against the R. microplus, P. humanus capitis, and the larvae of An. subpictus, Cx. quinquefasciatus with LC(50) values of 29.14, 11.80, 11.14, and 12.39 mg/L; r (2) = 0.805, 0.876, 0.894, and 0.904, respectively. The synthesized ZnO NPs showed the LC (50) and r (2) values against the R. microplus (13.41 mg/L; 0.982), P. humanus capitis (11.80 mg/L; 0.966), and the larvae of An. subpictus (3.19; 0.945 mg/L), against Cx. quinquefasciatus (4.87 mg/L; 0.970), respectively.

Table 2.

Efficacy of F. religiosa aqueous leaf extract, Syntheiszed ZnO NPs and TiO2 NPs against the An. stephensi larvae with their probit equation, LC50 with 95% CL, χ2, p and R2 values.

Instar Concentrations (ppm) % mortality Probit equation LC50 (±CL) χ2 p R2
Extract I 25 0 y = 0.2203x + 0.6641 250 ± 1.22 1.95 0.744 0.903
50 10
100 30
150 40
250 50
II 25 0 y = 0.2203x + 0.6641 250 ± 1.22 1.95 0.744 0.903
50 10
100 30
150 40
250 50
III 25 30 y = 0.1016x + 28.32 200 ± 1.18 2.27 0.686 0.825
50 30
100 40
150 50
250 50
IV 25 0 y = 0.3125x - 15.938 200 ± 1.14 1.54 0.820 0.919
50 20
100 30
150 50
250 60
ZnO NPs I 25 40 y = 0.2297x + 35.586 50 ± 0.23 2.67 0.615 0.898
50 50
100 60
150 70
250 80
II 25 40 y = 0.2297x + 35.586 75 ± 0.25 2.74 0.603 0.898
50 50
100 50
150 80
250 90
III 25 50 y = 0.1859x + 42.617 2 5 ± 0.23 2.74 0.602 0.988
50 50
100 60
150 70
250 90
IV 25 60 ** ** ** ** **
50 90
100 100
150 100
250 100
TiO2 NPs I 25 60 y = 0.1188x + 62.344 15 ± 0.12 2.912 0.573 0.868
50 70
100 80
150 80
250 90
II 25 40 y = 0.2141x + 37.383 50 ± 0.29 2.727 0.604 0.991
50 50
100 60
150 70
250 90
III 25 50 y = 0.1234x + 49.805 25 ± 0.23 2.689 0.611 0.938
50 60
100 60
150 70
250 80
IV 25 50 y = 0.1234x + 49.805 25 ± 0.34 2.710 0.608 0.938
50 60
100 60
150 70
250 80
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** 100% mortality.

The TiO2 NPs were found effective against the larvae of An. stephensi. The mortality was recorded after 48 h of exposure. The first instar (LC50 15 ppm), second instar (LC50 50 ppm), third instar (LC50 25 ppm) and fourth instar (LC50 25 ppm) larvae were observed with their probit equations and 95% confidential limit, R2, chi-square and p value after 24 h (Table 2). The larvicidal activity of titanium dioxide nanoparticles (TiO2 NPs) synthesized from the root aqueous extract of M. citrifolia against the larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus has been assessed (Suman et al., 2015). The biosynthesized TiO2 NPS showed maximum activity against the larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus when compared to the aqueous extract of M. citrifolia. Similarly, the anti-parasitic activity of TiO2 NPs against the larvae of R. microplus, H. anatolicum anatolicum and H. bispinosa, fourth instar larvae of An. subpictus, and Cx. quinquefasciatus has been assessed by (Rajakumar et al., 2015). The maximum efficacy was observed in synthesized TiO2 NPs against the larvae of R. microplus, H. anatolicum anatolicum, H. bispinosa, An. subpictus, and Cx. quinquefasciatus with LC value of 28.56, 33.17, 23.81, 5.84, and 4.34 mg/L, respectively. Recently, the larvicidal and the pediculicidal activity of synthesized titanium dioxide nanoparticles (TiO2 NPs) using the leaf aqueous extract of V. negundo against the fourth instar larvae of the malaria vector, An. subpictus Grassi and filariasis vector, Cx. quinquefasciatus Say and the head louse, P. humanus capitis De Geer has been carried out by (Gandhi et al., 2016). The maximum activity has been observed in the synthesized TiO2 NPs against An. subpictus, Cx. quinquefasciatus and lice, (LC50 = 7.52, 7.23 and 24.32 mg/L; χ2 = 0.161, 2.678 and 4.495; r2 = 0.663, 0.742 and 0.924), respectively. The larvicidal activity of synthesized ZnO and TiO2 has been reported by other researchers (Table 3)

Table 3.

Comparative larvicidal efficacy of synthesized ZnO and TiO2 nanoparticles against different mosquito species.

Plant species Common name Plant part used Test NPs tested Mosquito species References
Momordica charantia Bitter guard Leaf ZnO An. stephensi Cx. quinquefasciatus Gandhi et al., 2016
Syzgium cumini Black plum Seed ZnO Ae. aegypti Roopan et al., 2018
Scadoxus multiflorus Blood lily Leaf ZnO Ae. aegypti Abdullah Al-Dhabi and Valan Arasu, 2018
Morinda citrifolia Noni Root TiO2 An. stephensi Ae. aegypti, Cx. quinquefasciatus Suman et al., 2015
Mangifera indica Mango Leaf TiO2 An. stephensi Cx. quinquefasciatus Rajakumar et al., 2015
Vitex negundo Chinese chaste tree Leaf TiO2 An. stephensi Cx. quinquefasciatus Gandhi et al., 2016
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The larvae of An. stephensi have also shown the mortality against the aqueous leaves extract of F. religiosa and mortality was recorded after 48 h. The first instar (LC50 250 ppm), second instar (LC50 250 ppm), third instar (LC50 200 ppm) and fourth instar (LC50 200 ppm) larvae were observed with their probit equations and 95% confidential limit, R2, chi-square and p value after 24 h (Table 1). The larvicidal efficacy of different extracts of F. benghalensis against Cx. quinquefasciatus, Ae. aegypti and An. stephensi has been investigated (Govindarajan, 2010). The lethal concentration (LC50) values of F. benghalensis against early second, third and fourth larvae of Cx. quinquefasciatus, Ae. Aegypti and An. stephensi were 41.43, 58.21 and 74.32 ppm, 56.54, 70.29 and 80.85 ppm and 60.44, 76.41 and 89.55 ppm respectively. Further, the larvicidal efficacy of different solvent leaf extracts of F. benghalensis against Cx. tritaeniorhynchus and An. subpictus has been determined (Govindarajan et al., 2011). The LC50 and LC90 values of F. benghalensis against early third instar of Cx. tritaeniorhynchus and An. subpictus were 100.88, 159.76 ppm and 56.66, 85.84 ppm, respectively. Thereafter, the Larvicidal activity of Indian medicinal plants, C. berryi, C. pentandra, P. graveolens, T. peruviana, S.indicum, F. microcarpa, M. dubia, C. bonplandianus, F. religiosa and C. lacryma has been tested against Ae. aegypti mosquito. They found the highest LC50 values of methanol extracts of F. microcarpa against Ae. aegypti larvae were 91.63 ppm followed by, LC50 values of C. lacryma, P. graveolens, C. berryi, and M. dubia extracts against Ae. aegypti larvae were 92.77, 95.65, 96.52 and 100.12 ppm, respectively.

3.7. Antibacterial activity

The antibacterial assay for biologically synthesized ZnO and TiO2 NPs against the pathogens is shown in Fig. 8. Well diffusion method was used to provide the evidence for and validate the antibacterial activity of ZnO and TiO2 NPs against E. coli (Gram negative) and S. aureus, (Gram positive) bacteria. The antibacterial activity of the ZnO and TiO2 NPs was indicated by the formation of the zone. The diameter of the inhibition zone was measured in millimetre. The maximum zone of inhibition was observed against ZnO NPs in E. coli (8, 10, 12 and 14 mm) and S. aureus (6, 8, 10 and 12) (Table 4 and Fig. 8). Several research confirming antimicrobial activity of ZnO NPs against the food related bacteria B. subtilis, E. coli, P. fluorescens, S. typhimurium and S. aureus has been reported (Russell and Hugo, 1994; Ip et al., 2006). ZnO NPs are also known to exhibit antimicrobial activities against L. monocytogenes, S. enteritidis and E. coli (Russell and Hugo, 1994). The formation of hydrogen peroxide from the surface of ZnO is considered to be mainly responsible for its antimicrobial property (Rai et al., 2009).

Fig. 8.

Fig. 8

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Antibacterial activity of synthesized (a) ZnO NPs and (b) TiO2 NPs against E. coli and S. aureus.

Table 4.

Antibacterial activity of synthesized ZnO NPs and TiO2 NPs against E. coli and S. aureus.

Species
Zone of inhibition/mm
50 150 250 500
ZnO NPs E. coli 8 ± 0.612 10 ± 0.654 12 ± 0.712 14 ± 0.801
S. aureus 6 ± 0.563 8 ± 0.612 10 ± 0.654 12 ± 0.712
TiO2 NPs E. coli 7 ± 0.552 9 ± 0.642 10 ± 0.654 13 ± 0.752
S. aureus 5 ± 0.456 6 ± 0.563 8 ± 0.612 10 ± 0.654
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While, TiO2 NPs has shown the zone of inhibition in E. coli (7, 9, 10, and 13) and S. aureus (5, 6, 8 and 10) (Table 4 and Fig. 8). The higher zone of inhibition occurred at 500 ppm concentration of synthesized ZnO and TiO2 NPs. The antibacterial activity of TiO2 by pure plant extracts of B. variegata and T. cordifolia has been studied (Maurya et al., 2012). Plant extract/TiO2 nanocomposites have shown various level of antibacterial activity on different test microorganisms. The highest antibacterial potentiality expressed in terms of zone of inhibition (ZOI) in mm was exhibited by the aqueous extract of B. variegata /TiO2 (45 mm against E. faecalis and 30 mm against E. coli) and benzene extract of T. cordifolia /TiO2 (26 mm) nanocomposites. Similar results were reported (Kumar et al., 2014) using the biosynthesized and chemically synthesized titania nanoparticles and other researchers also (Table 5).

Table 5.

Comparative antibacterial activity of synthesized nanoparticles ZnO and TiO2 nanoparticles against different microorganisms.

Plant species Plant part used NPs tested Species References
Catharanthus roseus Leaf ZnO B. thuringiensis, E. coli, S. aureus, P. aeruginosa Bhumi and Savithramma, 2014
Brassica oleraceae Leaf ZnO E. coli, V. cholera, C. Botulinum, S. aureus, B. subtilis Raj et al., 2015
Trifolium pratense Flower ZnO S. aureus, P. aeruginosa, E. coli Dobrucka and Dugaszewska, 2016
Bauhinia tomentosa Leaf ZnO B. thuringiensis, E. coli, S. aureus, P. aeruginosa Sharmila et al., 2018
Pandanus odorifer Leaf ZnO B. subtilis, E. coli Hussain et al., 2019
Aloe vera Leaf ZnO E. coli Izwanie Rasli et al., 2020
Psidium guajava Leaf TiO2 E. coli, S. aureus Santhoshkumar et al., 2014
Cynodon dactylon Leaf TiO2 E. coli Hariharan et al., 2017
Glycosmis Leaf TiO2 S. saprophyticus, B. subtilis, E. coli, cochinchinensis P. aeruginosa Rosi and Kalyanasundaram, 2018
Cassia fistula Leaf TiO2 E. coli, S. aureus Swathi et al., 2019
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4. Conclusion

The present study, synthesis of ZnO NPs and TiO2 NPs from the F. religiosa is a green, rapid, cost-effective, non-toxic and eco-friendly approach for mosquito control as well as antimicrobial agent also.

Declaration of Competing Interest

Authors declare that there is no conflict of interest.

Acknowledgements

I sincerely grateful to Department of Science & Technology, Science and Engineering Research Board, New Delhi, providing the Young Scientist Project (File No.-DST SERB/SB/YS/LS-35/2014). I thank to Director ICMR-National Institute of Malaria Research, New Delhi for providing the space to run the project. I wish to thank Dr. T.C. Nagpal, AIIMS, New Delhi for TEM facility, Director CSIR-National Physical Laboratory, New Delhi for XRD and SEM-EDS facility.

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