Green synthesis of silver nanoparticles using Murraya koenigii leaf extract against Anopheles stephensi and Aedes aegypti

Mosquitoes transmit serious human diseases, causing millions of deaths every year. The use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of synthesized natural products for vector control have been a priority in this area. In the present study, the activity of silver nanoparticles (AgNPs) synthesized using Murraya koenigii plant leaf extract against first to fourth instars larvae and pupae of Anopheles stephensi and Aedes aegypti was determined. Range of concentrations of synthesized AgNPs (5, 10, 20, 30, and 40 ppm) and ethanol leaf extract (50, 200, 350, 500, and 650 ppm) were tested against the larvae of A. stephensi and A. aegypti. The synthesized AgNPs from M. koenigii leaf were highly toxic than crude leaf ethanol extract in both mosquito species. The results were recorded from UV–Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy analysis. Larvae were exposed to varying concentrations of aqueous extract of synthesized AgNPs for 24 h. The maximum mortality was observed in synthesized AgNPs, and ethanol leaf extract of M. koenigii against A. stephensi had LC₅₀ values of 10.82, 14.67, 19.13, 24.35, and 32.09 ppm and 279.33, 334.61, 406.95, 536.11, and 700.16 ppm and LC₉₀ values of 32.38, 42.52, 53.65, 63.51, and 75.26 ppm and 737.37, 843.84, 907.67, 1,187.62, and 1,421.13 ppm. A. aegypti had LC₅₀ values of 13.34, 17.19, 22.03, 27.57, and 34.84 ppm and 314.29, 374.95, 461.01, 606.50, and 774.01 ppm and LC₉₀ values of 36.98, 47.67, 55.95, 67.36, and 77.72 ppm and 777.32, 891.16, 1,021.90, 1,273.06, and 1,509.18 ppm, respectively. These results suggest that the use of M. koenigii synthesized silver nanoparticles can be a rapid, environmentally safer biopesticide which can form a novel approach to develop effective biocides for controlling the target vector mosquitoes.     

Chemical constituents and larvicidal potential of Feronia limonia leaf essential oil against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus

In the present investigation, the leaf essential oil of Feronia limonia was evaluated for chemical constituents and mosquito larvicidal activity against the larvae of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus. GC and GC–MS analyses revealed that the essential oil contain 51 compounds. Estragole (34.69 %) and β-pinene(23.59 %) were identified as the major constituents followed by methyl (Z)-caryophyllene (11.05 %), eugenol (6.50 %), linalool (3.97 %), phytol (3.27 %), sabinene (2.41 %) and limonene (2.27 %). Larval mortality was observed after 12 and 24 h of exposure period. The oil showed remarkable larvicidal activity against A. stephensi (LC₅₀?=?38.93 and LC₉₀?=?108.64 ppm (after 12 h); LC₅₀?=?15.03 and LC₉₀?=?36.69 ppm (after 24 h)), A. aegypti (LC₅₀?=?37.60 and LC₉₀?=?104.69 ppm (after 12 h); LC₅₀?=?11.59 and LC₉₀?=?42.95 ppm (after 24 h)) and C. quinquefasciatus (LC₅₀?=?52.08 and LC₉₀?=?124.33 ppm (after 12 h); LC₅₀?=?22.49 and LC₉₀?=?60.90 ppm (after 24 h)). Based on the results, the essential oil of F. limonia can be considered as a new source of larvicide for the control of vector mosquitoes.     

Mosquito larvicidal, pupicidal, adulticidal, and repellent activity of Artemisia nilagirica (Family: Compositae) against Anopheles stephensi and Aedes aegypti

Mosquito-borne diseases have an economic impact, including loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. The aim of the present study, to evaluate the larvicidal, pupicidal, repellent, and adulticidal activities of methanol crude extract of Artemisia nilagirica were assayed for their toxicity against two important vector mosquitoes, viz., Anopheles stephensi and Aedes aegypti (Diptera: Culicidae). The fresh leaves of A. nilagirica were washed thoroughly in tap water and shade dried at room temperature (28?±?2?°C) for 5?C8?days. The air-dried materials were powdered separately using commercial electrical blender. From the plants, 500?g powdered was macerated with 1.5?L organic solvents of methanol sequentially for a period of 72?h each and filtered. The larval and pupal mortality was observed after 24?h of exposure; no mortality was observed in the control group. The first- to fourth-instar larvae and pupae of A. stephensi had values of LC₅₀?=?272.50, 311.40, 361.51, 442.51, and 477.23?ppm, and the LC₉₀?=?590.07, 688.81, 789.34, 901.59, and 959.30?ppm; the A. aegypti had values of LC₅₀?=?300.84, 338.79, 394.69, 470.74, and 542.11?ppm, and the LC₉₀?=?646.67, 726.07, 805.49, 892.01, and 991.29?ppm, respectively. The results of the repellent activity of plant extract of A. nilagirica plants at five different concentrations of 50, 150, 250, 350, and 450?ppm were applied on skin of fore arm in man and exposed against adult female mosquitoes. In this observation, the plant crude extract gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. The adult mortality was found in methanol extract of A. nilagirica, with the LC₅₀ and LC₉₀ values of 205.78 and 459.51?ppm for A. stephensi, and 242.52 and 523.73?ppm for A. aegypti, respectively. This result suggests that the leaf extract have the potential to be used as an ideal eco-friendly approach for the control of vector mosquito as target species.     

Low-cost and eco-friendly green synthesis of silver nanoparticles using Feronia elephantum (Rutaceae) against Culex quinquefasciatus,Anopheles stephensi,and Aedes aegypti (Diptera: Culicidae)

Mosquitoes transmit serious human diseases, causing millions of deaths every year. The use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of synthesized natural products for vector control have been a priority in this area. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Feronia elephantum plant leaf extract against late third-instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus was determined. The range of concentrations of synthesized AgNPs (5, 10, 15, 20, and 25 μg mL^?1) and aqueous leaf extract (25, 50, 75, 100, and 125 μg mL^?1) were tested against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of F. elephantum for all three important vector mosquitoes. The synthesized AgNPs from F. elephantum were highly toxic than crude leaf aqueous extract to three important vector mosquito species. The results were recorded from UV–visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy analysis (EDX). Synthesized AgNPs against the vector mosquitoes A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC₅₀ and LC₉₀ values: A. stephensi had LC₅₀ and LC₉₀ values of 11.56 and 20.56 μg mL^?1; A. aegypti had LC₅₀ and LC₉₀ values of 13.13 and 23.12 μg mL^?1; and C. quinquefasciatus had LC₅₀ and LC₉₀ values of 14.19 and 24.30 μg mL^?1. No mortality was observed in the control. These results suggest that the green synthesis of silver nanoparticles using F. elephantum has the potential to be used as an ideal eco-friendly approach for the control of A. stephensi, A. aegypti, and C. quinquefasciatus. This is the first report on the mosquito larvicidal activity of the plant extracts and synthesized nanoparticles.     

Mosquito larvicidal properties of silver nanoparticles synthesized using Heliotropium indicum (Boraginaceae) against Aedes aegypti,Anopheles stephensi,and Culex quinquefasciatus (Diptera: Culicidae)

Mosquitoes transmit dreadful diseases to human beings wherein biological control of these vectors using plant-derived molecules would be an alternative to reduce mosquito population. In the present study activity of aqueous leaf extract and silver nanoparticles (AgNPs) synthesized using Helitropium indicum plant leaves against late third instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. The range of varying concentrations of synthesized AgNPs (8, 16, 24, 32, and 40 μg/mL) and aqueous leaf extract (30, 60, 90, 120, and 150 μg/mL) were tested against the larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The synthesized AgNPs from H. indicum were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV–Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy analysis, transmission electron microscopy, and histogram. The synthesized AgNPs showed larvicidal effects after 24 h of exposure. Considerable mortality was evident after the treatment of H. indicum for all three important vector mosquitoes. The LC₅₀ and LC₉₀ values of H. indicum aqueous leaf extract appeared to be effective against A. stephensi (LC₅₀, 68.73 μg/mL; LC₉₀, 121.07 μg/mL) followed by A. aegypti (LC₅₀, 72.72 μg/mL; LC₉₀, 126.86 μg/mL) and C. quinquefasciatus (LC₅₀, 78.74 μg/mL; LC₉₀, 134.39 μg/mL). Synthesized AgNPs against the vector mosquitoes of A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC₅₀ and LC₉₀ values: A. stephensi had LC₅₀ and LC₉₀ values of 18.40 and 32.45 μg/mL, A. aegypti had LC₅₀ and LC₉₀ values of 20.10 and 35.97 μg/mL, and C. quinquefasciatus had LC₅₀ and LC₉₀ values of 21.84 and 38.10 μg/mL. No mortality was observed in the control. These results suggest that the leaf aqueous extracts of H. indicum and green synthesis of silver nanoparticles have the potential to be used as an ideal ecofriendly approach for the control of A. stephensi, A. aegypti, and C. quinquefasciatus. This is the first report on the mosquito larvicidal activity of the plant extracts and synthesized nanoparticles.     

Bioefficacy of larvicdial and pupicidal properties of Carica papaya (Caricaceae) leaf extract and bacterial insecticide, spinosad, against chikungunya vector, Aedes aegypti (Diptera: Culicidae)

The present study was carried out to establish the properties of Carica papaya leaf extract and bacterial insecticide, spinosad on larvicidal and pupicidal activity against the chikungunya vector, Aedes aegypti. The medicinal plants were collected from the area around Bharathiar University, Coimbatore, India. C. papaya leaf was washed with tap water and shade-dried at room temperature. An electrical blender powdered the dried plant materials (leaves). The powder (500 g) of the leaf was extracted with 1.5 l of organic solvents of methanol for 8 h using a Soxhlet apparatus and then filtered. The crude leaf extracts were evaporated to dryness in a rotary vacuum evaporator. The plant extract showed larvicidal and pupicidal effects after 24 h of exposure; however, the highest larval and pupal mortality was found in the leaf extract of methanol C. papaya against the first- to fourth-instar larvae and pupae of values LC₅₀ = I instar was 51.76 ppm, II instar was 61.87 ppm, III instar was 74.07 ppm, and IV instar was 82.18 ppm, and pupae was 440.65 ppm, respectively, and bacterial insecticide, spinosad against the first to fourth instar larvae and pupae of values LC₅₀ = I instar was 51.76 ppm, II instar was 61.87 ppm, III instar was 74.07 ppm, and IV instar was 82.18 ppm, and pupae was 93.44 ppm, respectively. Moreover, combined treatment of values of LC₅₀ = I instar was 55.77 ppm, II instar was 65.77 ppm, III instar was 76.36 ppm, and IV instar was 92.78 ppm, and pupae was 107.62 ppm, respectively. No mortality was observed in the control. The results that the leaves extract of C. papaya and bacterial insecticide, Spinosad is promising as good larvicidal and pupicidal properties of against chikungunya vector, A. aegypti. This is an ideal eco-friendly approach for the control of chikungunya vector, A. aegypti as target species of vector control programs.     

Green synthesis of silver nanoparticles using Sida acuta (Malvaceae) leaf extract against Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti (Diptera: Culicidae)

Mosquitoes act as a vector for most of the life-threatening diseases like malaria, yellow fever, dengue fever, chikungunya fever, filariasis, encephalitis, West Nile Virus infection, etc. Under the Integrated Mosquito Management, emphasis was given on the application of alternative strategies in mosquito control. The continuous application of synthetic insecticides causes development of resistance in vector species, biological magnification of toxic substances through the food chain, and adverse effects on environmental quality and nontarget organisms including human health. Application of active toxic agents from plant extracts as an alternative mosquito control strategy was available from ancient times. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Sida acuta plant leaf extract against late third instar larvae of Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti was determined. Range of concentrations of synthesized AgNPs (10, 20, 30, 40, and 50 μg/mL) and aqueous leaf extract (50, 100, 150, 200, and 250 μg/mL) were tested against the larvae of C. quinquefasciatus, A. stephensi and A. aegypti. The synthesized AgNPs from S. acuta leaf were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV–Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of S. acuta for all three important vector mosquitoes. The LC₅₀ and LC₉₀ values of S. acuta aqueous leaf extract appeared to be most effective against A. stephensi (LC₅₀, 109.94 μg/mL and LC₉₀, 202.42 μg/mL) followed by A. aegypti LC₅₀ (119.32 μg/mL and LC₉₀, 213.84 μg/mL) and C. quinquefasciatus (LC₅₀, 130.30 μg/mL and LC₉₀, 228.20 μg/mL). Synthesized AgNPs against the vector mosquitoes of A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC₅₀ and LC₉₀ values: A. stephensi had LC₅₀ and LC₉₀ values of 21.92, and 41.07 μg/mL; A. aegypti had LC₅₀ and LC₉₀ values of 23.96, and 44.05 μg/mL; C. quinquefasciatus had LC₅₀ and LC₉₀ values of 26.13 and 47.52 μg/mL. These results suggest that the use of S. acuta synthesized silver nanoparticles can be a rapid, environmentally safer biopesticide which can form a novel approach to develop effective biocides for controlling the target vector mosquitoes. This is the first report on the mosquito larvicidal activity of the plant aqueous extract and synthesized nanoparticles.     

Ovicidal, repellent, adulticidal and field evaluations of plant extract against dengue, malaria and filarial vectors

Mosquitoes are insect vectors responsible for the transmission of parasitic and viral infections to millions of people worldwide, with substantial morbidity and mortality. Infections transmitted by mosquitoes include malaria, yellow fever, chikungunya, filariasis and other arboviruses. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. The adulticidal activities of crude hexane, benzene, ethyl acetate, acetone and methanol leaf extracts of Acalypha alnifolia were assayed for their toxicity against three important vector mosquitoes, viz., Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. The adult mortality was observed after 24 h of exposure. All extracts showed moderate adulticide effects; however, the highest adult mortality was found in methanol extract were observed. The LC₅₀ values of A. alnifolia leaf extracts against adulticidal activity of (hexane, benzene, ethyl acetate, acetone and methanol) A. aegypti, A. stephensi and C. quinquefasciatus were the following: A. aegypti values were 371.87, 342.97, 320.17, 300.86 and 279.75 ppm; A. stephensi values were 358.35, 336.64, 306.10, 293.01 and 274.76 ppm; C. quinquefasciatus values were 383.59, 354.13, 327.74, 314.33 and 291.71 ppm. The results of the repellent activity of hexane, benzene, ethyl acetate, acetone and methanol extract of A. alnifolia plant at three different concentrations of 1.0, 3.0, and 5.0 mg/cm² were applied on skin of forearm in man and exposed against adult female mosquitoes. In this observation, this plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. Mean percent hatchability of the ovicidal activity was observed 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Mortality of 100 % with methanol extract of A. alnifolia was exerted at 125 and 300 ppm. The larval density was decreased after the treatment of plant extracts at the breeding sites (water bodies system) of vector mosquitoes, and hence, these plant extracts are suitable alternatives of synthetic insecticides for mosquito vector management.These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes. This study provides first report on the mosquito ovicidal, repellent and adulticidal activities of these plant extracts against mosquito vector species from India.     

Larvicidal,ovicidal, and adulticidal efficacy of Erythrina indica (Lam.) (Family: Fabaceae) against Anopheles stephensi,Aedes aegypti,and Culex quinquefasciatus (Diptera: Culicidae)

Mosquitoes are the major vector for the transmission of malaria, dengue fever, yellow fever, filariasis, schistosomiasis, and Japanese encephalitis. Mosquito control is facing a threat because of the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal, ovicidal, and adulticidal potential of the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts from the medicinal plant Erythrina indica against the medically important mosquito vectors, Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of leaf of E. indica against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus with the LC₅₀ and LC₉₀ values of 69.43, 75.13, and 91.41 ppm and 125.49, 134.31, and 167.14 ppm, respectively. The mean percent hatchability of the eggs was observed after 48 h post treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. The methanol extract of E. indica against A. stephensi, A. aegypti, and C. quinquefasciatus exerted 100 % mortality (zero hatchability) at 150, 200, and 250 ppm, respectively. Control eggs showed above 99.3–100 % hatchability. The adult mortality was observed after 24 h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in methanol extract against A. stephensi followed by A. aegypti and C. quinquefasciatus with the LD₅₀ and LD₉₀ values of 88.76, 94.09, and 119.64 ppm and 160.83, 169.01, and 219.77 ppm, respectively. No mortality was recorded in the control. Our data suggest that the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts of E. indica have the potential to be used as an eco-friendly approach for the control of the A. stephensi, A. aegypti, and C. quinquefasciatus.     

Evaluation of leaf aqueous extract and synthesized silver nanoparticles using Nerium oleander against Anopheles stephensi (Diptera: Culicidae)

Green nanoparticle synthesis has been achieved using environmentally acceptable plant extract and ecofriendly reducing and capping agents. The present study was carried out to establish the larvicidal activity of synthesized silver nanoparticles (AgNPs) using leaf extract of Nerium oleander (Apocynaceae) against the first to fourth instar larvae and pupae of malaria vector, Anopheles stephensi (Diptera: Culicidae). Nanoparticles are being used in many commercial applications. It was found that aqueous silver ions can be reduced by the aqueous extract of the plant parts to generate extremely stable silver nanoparticles in water. The results were recorded from UV–Vis spectrum, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and energy-dispersive X-ray (EDX) spectroscopy analysis. The production of the AgNPs synthesized using leaf extract of N. oleander was evaluated through a UV–Vis spectrophotometer in a wavelength range of 200 to 700 nm. This revealed a peak at 440 nm in N. oleander leaf extracts, indicating the production of AgNPs. The FTIR spectra of AgNPs exhibited prominent peaks at 509.12 cm^?1 (C–H bend alkenes), 1,077.05 cm^?1 (C–O stretch alcohols), 1,600.63 cm^?1 (N–H bend amines), 2,736.49 and 2,479.04 cm^?1 (O–H stretch carboxylic acids), and 3,415.31 cm^?1 (N–H stretching due to amines group). An SEM micrograph showed 20–35-nm-size aggregates of spherical- and cubic-shaped nanoparticles. EDX showed the complete chemical composition of the synthesized nanoparticles of silver. Larvicidal activity of aqueous leaf extract of N. oleander and synthesized AgNPs was carried out against Anopheles stephensi, and the results showed that the highest larval mortality was found in the synthesized AgNPs against the first to fourth instar larvae and pupae of Anopheles stephensi with the following values: LC₅₀ of instar larvae 20.60, 24.90, 28.22, and 33.99 ppm; LC₉₀ of instar larvae 41.62, 50.33, 57.78, and 68.41 ppm; and LC₅₀ and LC₉₀ of pupae 39.55 and 79.10 ppm, respectively. The aqueous leaf extract exhibited larval toxicity against the first to fourth instar larvae and pupae of Anopheles stephensi with the following values: LC₅₀ of instar larvae 232.90, 273.71, 318.94, and 369.96 ppm; LC₉₀ of instar larvae 455.95, 563.10, 639.86, and 730.30 ppm; and LC₅₀ and LC₉₀ of pupae 426.01 and 805.13 ppm, respectively. The chi-square value was significant at p?<?0.05 level. The possible larvicidal activity may be due to penetration of nanoparticles through a membrane. The results could suggest that the use of plant N. oleander to synthesize silver nanoparticles is a rapid, environmentally safer, and greener approach for mosquito control. This could lead us to a new possibility in vector-control strategy.     

Entomopathogenic marine actinomycetes as potential and low-cost biocontrol agents against bloodsucking arthropods

A novel approach to control strategies for integrated blood-feeding parasite management is in high demand, including the use of biological control agents. The present study aims to determine the efficacy of optimized crude extract of actinomycetes strain LK1 as biological control agent against the fourth-instar larvae of Anopheles stephensi and Culex tritaeniorhynchus (Diptera: Culicidae) and adults of Haemaphysalis bispinosa, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), and Hippobosca maculata (Diptera: Hippoboscidae). Antiparasitic activity was optimized using the Plackett–Burman method, and the design was developed using the software Design-Expert version 8.0.7.1. The production of the optimized crude actinomycetes LK1 strain extract was performed using response surface methodology to optimize the process parameters of protease inhibitor activity of marine actinobacteria for the independent variables like pH, temperature, glucose, casein, and NaCl at two levels (?1 and +1). The potential actinomycetes strain was identified as Saccharomonas spp., and the metamodeling surface simulation procedure was followed. It was studied using a computer-generated experimental design, automatic control of simulation experiments, and sequential optimization of the metamodels fitted to a simulation response surface function. The central composite design (CCD) used for the analysis of treatment showed that a second-order polynomial regression model was in good agreement with the experimental results at R ²?=?0.9829 (p?<?0.05). The optimized values of the variables for antioxidant production were pH 6.00, glucose 1.3 %, casein 0.09 %, temperature 31.23 °C, and NaCl 0.10 %. The LK1 strain-optimized crude extract was purified using reversed-phase high-pressure liquid chromatography, and the isolated protease inhibitor showed antiparasitic activity. The antiparasitic activity of optimized crude extract of LK1 was tested against larvae of A. stephensi (LC₅₀?=?31.82 ppm; r ²?=?0.818) and C. tritaeniorhynchus (LC₅₀?=?26.62 ppm; r ²?=?0.790) and adults of H. bispinosa (LC₅₀?=?106.58 ppm; r ²?=?0.871), R. (B.) microplus (LC₅₀?=?92.96 ppm; r ²?=?0.913), and H. maculata (LC₅₀?=?84.90 ppm; r ²?=?0.857).     

Silver nanoparticles: a possibility for malarial and filarial vector control technology

Green synthesis technology is one of the rapid, reliable and best routes for the synthesis of silver nanoparticles (AgNPs). There are bioactive compounds with enormous potential in Azadirachta indica (Neem). The extraordinary mosquitoes warrant nanotechnology to integrate with novel molecules. This will be sustainable technology for future. Here, we synthesized AgNPs using aqueous extracts of leaves and bark of Az. indica (Neem). We tested AgNPs as larvicides, pupicides and adulticides against the malaria vector Anopheles stephensi and filariasis vector Culex quinquefasciatus. The results were obtained using UV-visible spectrophotometer and the images were recorded with a transmission electron microscope (TEM). The efficacy tests were then performed at different concentrations varying many hours by probit analysis. The synthesized AgNPs were spherical in shape and with varied sizes (10.47-nm leaf and 19.22-nm bark). The larvae, pupae and adults of filariasis vector C. quinquefasciatus were found to be more susceptible to our AgNPs than the malaria vector An. stephensi. The first and the second instar larvae of C. quinquefasciatus show a mortality rate of 100 % after 30 min of exposure. The results against the pupa of C. quinquefasciatus were recorded as LC₅₀ 4 ppm, LC₉₀ 11 ppm and LC₉₉ 13 ppm after 3 h of exposure. In the case of adult mosquitoes, LC₅₀ 1.06 μL/cm², LC₉₀ 2.13 μL/cm² and LC₉₉ 2.4 μL/cm² were obtained after 4 h of exposure. These results suggest that our AgNPs are environment-friendly for controlling malarial and filarial vectors.     

Strong larvicidal potential of Artemisia annua leaf extract against malaria (Anopheles stephensi Liston) and dengue (Aedes aegypti L.) vectors and bioassay-driven isolation of the marker compounds

Malaria and dengue are the two most important vector-borne human diseases caused by mosquito vectors Anopheles stephensi and Aedes aegypti, respectively. Of the various strategies adopted for eliminating these diseases, controlling of vectors through herbs has been reckoned as one of the important measures for preventing their resurgence. Artemisia annua leaf chloroform extract when tried against larvae of A. stephensi and A. aegypti has shown a strong larvicidal activity against both of these vectors, their respective LC50 and LC90 values being 0.84 and 4.91 ppm for A. stephensi and 0.67 and 5.84 ppm for A. aegypti. The crude extract when separated through column chromatography using petroleum ether-ethyl acetate gradient (0–100 %) yielded 76 fractions which were pooled into three different active fractions A, B and C on the basis of same or nearly similar R _f values. The aforesaid pooled fractions when assayed against the larvae of A. stephensi too reported a strong larvicidal activity. The respective marker compound purified from the individual fractions A, B and C, were Artemisinin, Arteannuin B and Artemisinic acid, as confirmed and characterized through FT-IR and NMR. This is our first report of strong mortality of A. annua leaf chloroform extract against vectors of two deadly diseases. This technology can be scaled up for commercial exploitation.     

Larvicidal activity of silver nanoparticles synthesized using <Emphasis Type="Italic">Plumeria rubra</Emphasis> plant latex against <Emphasis Type="Italic">Aedes aegypti</Emphasis> and <Emphasis Type="Italic">Anopheles stephensi</Emphasis>

In the present study activity of silver nanoparticles (AgNPs) synthesized using Plumeria rubra plant latex against second and fourth larval instar of Aedes aegypti and Anopheles stephensi was determined. Range of concentrations of synthesized AgNps (10, 5, 2.5, 1.25, 0.625, 0.3125 ppm) and aqueous crude latex (1,000, 500, 250, 125, 62.50, 31.25 ppm) were tested against larvae of A. aegypti and A. Stephensi. The synthesized AgNps from P. rubra latex were highly toxic than crude latex extract in both mosquito species. The LC₅₀ values for second and fourth larval instars after 24 h of crude latex exposure were 1.49, 1.82 ppm against A. aegypti and 1.10, 1.74 ppm against A. stephensi respectively. These figures were 181.67, 287.49 ppm against A. aegypti and 143.69, 170.58 ppm against A. stephensi respectively for crude latex extract. The mortality rates were positively correlated with the concentration of AgNPs. The characterization studies of synthesized AgNPs by UV–Vis spectrophotometry, transmission electron microscopy (TEM), Particle size analysis (PSA) and zeta potential confirmed the spherical shape and size (32–200 nm) of silver nanoparticles alongwith stability. Toxicity studies carried out against non-target fish species Poecilia reticulata, the most common organism in the habitats of A. aegypti and A. stephensi showed no toxicity at LC₅₀ and LC₉₀ doses of the AgNPs. This is the first report on mosquito larvicidal activity of latex synthesized nanoparticles.     

Ovicidal,larvicidal and adulticidal properties of Asparagus racemosus (Willd.) (Family: Asparagaceae) root extracts against filariasis (Culex quinquefasciatus), dengue (Aedes aegypti) and malaria (Anopheles stephensi) vector mosquitoes (Diptera: Culicidae)

Several diseases are associated to the mosquito–human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present investigation was undertaken to study the ovicidal, larvicidal and adulticidal activities of crude hexane, ethyl acetate, benzene, chloroform and methanol extracts of root of Asparagus racemosus were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. The methanol extract of Asparagus racemosus against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi exerted 100 % mortality (zero hatchability) at 375, 300 and 225 ppm, respectively. Control eggs showed 99–100 % hatchability. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of root of Asparagus racemosus against the larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi with the LC₅₀ and LC₉₀ values were 115.13, 97.71 and 90.97 ppm and 210.96, 179.92, and 168.82 ppm, respectively. The adult mortality was observed after 24 h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in methanol extract against Anopheles stephensi followed by Aedes aegypti and Culex quinquefasciatus with the LD₅₀ and LD₉₀ values were 120.44, 135.60, and 157.71 ppm and 214.65, 248.35, and 290.95 ppm, respectively. No mortality was recorded in the control. The finding of the present investigation revealed that the root extract of Asparagus racemosus possess remarkable ovicidal, larvicidal and adulticidal activity against medically important vector mosquitoes and this is the low cost and ideal eco-friendly approach for the control of mosquitoes. This is the first report on the mosquito ovicidal, larvicidal and adulticidal activities of the reported Asparagus racemosus root.     

Studies on the impact of biosynthesized silver nanoparticles (AgNPs) in relation to malaria and filariasis vector control against Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae)

Biosynthesized nanoparticles have been achieved using environmentally acceptable plant extract and eco-friendly reducing and capping agents. The present study was based on assessments of the larvicidal activities to determine the efficacies of synthesized silver nanoparticles (AgNPs) using aqueous leaf extract of Vinca rosea (L.) (Apocynaceae) against the larvae of malaria vector Anopheles stephensi Liston and filariasis vector Culex quinquefasciatus Say (Diptera: Culicidae). Larvae were exposed to varying concentrations of aqueous extract of V. rosea and synthesized AgNPs for 24, 48, and 72 h. AgNPs were rapidly synthesized using the leaf extract of V. rosea, and the formation of nanoparticles was observed within 15 min. The results recorded from UV–Vis spectrum, Fourier transform infrared (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) support the biosynthesis and characterization of AgNPs. The formation of the AgNPs synthesized from the XRD spectrum compared with the Bragg reflections at 2θ?=?29.36, 38.26, 44.51, 63.54, and 77.13° which can be indexed to the (121), (111), (200), (220), and (311) orientations, respectively, confirmed the presence of AgNPs. The FTIR spectra of AgNPs exhibited prominent peaks at the spectra showed sharp and strong absorption band at 3,406.71 to 3,431.90 cm^?1 double in case of NH₂ group of a primary amine (N–H stretch). The presence of the sharp peak at 2,926.54 to 2,925.80 cm^?1 very broad often looks like distorted baseline (O–H carboxylic acids). The band 1,633.26 to 1,625.81 cm^?1 was assigned to C?=?C alkenes, aromatic ring stretching vibration, respectively. SEM analysis of the synthesized AgNPs clearly showed the clustered and irregular shapes, mostly aggregated and having the size of 120 nm. TEM reveals spherical shape of synthesized AgNPs. Particle size analysis revealed that the size of particles ranges from 25 to 47 nm with average size of 34.61 nm. Energy-dispersive X-ray spectroscopy showed the complete chemical composition of the synthesized AgNPs. In larvicidal activity, the results showed that the maximum efficacy was observed in synthesized AgNPs against the fourth instar larvae of A. stephensi (LC₅₀?=?12.47 and 16.84 mg/mL and LC₉₀?=?36.33 and 68.62 mg/mL) on 48 and 72 h of exposure and against C. quinquefasciatus (LC₅₀?=?43.80 mg/mL and LC₉₀?=?120.54 mg/mL) on 72-h exposure, and aqueous extract showed 100 % mortality against A. stephensi and C. quinquefasciatus (LC₅₀?=?78.62 and 55.21 mg/mL and LC₉₀?=?184.85 and 112.72 mg/mL) on 72-h exposure at concentrations of 50 mg/mL, respectively. The AgNPs did not exhibit any noticeable toxicity on Poecilia reticulata after 24, 48, and 72 h of exposure. These results suggest that the synthesized AgNPs have the potential to be used as an ideal eco-friendly approach for the control of the A. stephensi and C. quinquefasciatus. This method is considered as a new approach to control vectors. Therefore, this study provides the first report on the mosquito larvicidal activity of V. rosea synthesized AgNPs against vectors.     

Effect of bioactive fractions of <Emphasis Type="Italic">Citrullus vulgaris</Emphasis> Schrad. leaf extract against <Emphasis Type="Italic">Anopheles stephensi</Emphasis> and <Emphasis Type="Italic">Aedes aegypti</Emphasis>

The benzene extract of Citrullus vulgaris was tested against Anopheles stephensi and Aedes aegypti for the larvicidal activity and ovicidal properties. The crude benzene extract was found to be more effective against A. stephensi than A. aegypti. The LC₅₀ values were 18.56 and 42.76 ppm respectively. The LC₅₀ values for silica gel fractions (bioactive fractions I, II, III and IV) were 11.32, 14.12, 14.53 and 16.02 ppm respectively. The mean per cent hatchability of the egg rafts were observed after 48 h post treatment. The crude extract of benzene exerted 100% mortality at 250 ppm against A. stephensi and at 300 ppm against A. aegypti. The silica gel fractions I and II afforded 100% mortality at 100 ppm and III and IV exerted the hatchability rate of 4.9 and 5.3% at the same concentration against A. stephensi.     

Knockdown and larvicidal activity of six monoterpenes against Aedes aegypti (Diptera: Culicidae) and their structure-activity relationships

The relationships between physicochemical parameters of majority components of Eucalyptus essential oils and their insecticide effect were evaluated on Aedes aegypti (L.) (Diptera: Culicidae). The octanol–water partition coefficients of the monoterpenes were estimated by the atom/fragment contribution method and the vapor pressures were determined by our laboratory in previous studies. The larvicidal activity (LC_{50 (ppm)}) and knockdown effect (KT_{50 (min)}) of each component was determined. The results show that the toxicity of EOs main components of Eucalyptus on adults and larvae of A. aegypti is strongly related to their physicochemical properties (vapor pressure and Log P). However, the interaction of both variables (vapor pressure * Log P) explains the toxicological phenomenon more precisely. The regression models were expressed as follows: KT _50(min)?=????10.9?+?3.7?*?Log P?+?1.9?*?1/P_vapor (R²?=?0.80;?F?=?42.5) and LC _50(ppm)?=????94.3?+?438.6?*? 1/Log P?+?2.8?*? 1/P_vapor (F?=?57.8;?R²?=?0.85). The six evaluated components present different functional groups. Therefore, it was considered to evaluate the monoterpenes as a group and separated in two groups: oxygenated monoterpenes (α-terpineol, 4-terpineol, and 1,8-cineole) and terpene hydrocarbons (γ-terpinene, p-cymene, and α-pinene). The results show the regression models for each group as follows: (A) oxygenated terpenes: KT _50(min)?=???515.3?+?1613.2???1/Log P?+?5,?2???1/P_vapor (F?=?3176.7?R²?=?0.99) and LC _50(ppm) ?=????1679.4?+?5402.1???1/Log P?+?12.7??? 1/P_vapor (F?=?282.9;?R²?=?0.99). (B) Hydrocarbons terpenes: KT _50(min)?=?18.2???58.3?*?1/Log P?+?2.7?*?1/P_vapor (F?=?171.7;? R²?=?0.97) and LC _50(ppm)?=???21.1?+?174.9???1/Log P???14.3???1/P_vapor (F?=?410.0;?R²?=?0.99). The association between the toxic effect of the evaluated monoterpenes against A. aegypti and the physicochemical properties can be better described when they are separated into functional groups (hydrocarbons vs. oxygenated terpenes).     

Adulticidal and repellent properties of Cassia tora Linn. (Family: Caesalpinaceae) against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi

Mosquitoes have developed resistance to various synthetic insecticides, making its control increasingly difficult. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. The adulticidal and repellent activities of crude hexane, chloroform, benzene, acetone, and methanol extracts of the leaf of Cassia tora were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. The adult mortality was observed after 24?h of exposure. All extracts showed moderate adulticidal effects; however, the highest adult mortality observed was found in methanol extract. The LC₅₀ and LC₉₀ values of C. tora leaf extracts against adulticidal activity of (hexane, chloroform benzene, acetone, and methanol) C. quinquefasciatus, A. aegypti, and A. stephensi were the following: C. quinquefasciatus LC₅₀ values were 338.81, 315.73, 296.13, 279.23, and 261.03?ppm and LC₉₀ values were 575.77, 539.31, 513.99, 497.06, and 476.03?ppm; A. aegypti LC₅₀ values were 329.82, 307.31, 287.15, 269.57, and 252.03?ppm and LC₉₀ values were 563.24, 528.33, 496.92, 477.61, and 448.05?ppm; and A. stephensi LC₅₀ values were 317.28, 300.30, 277.51, 263.35, and 251.43?ppm and LC₉₀ values were 538.22, 512.90, 483.78, 461.08, and 430.70?ppm, respectively. The results of the repellent activity of hexane, chloroform, benzene, acetone, and methanol extracts of C. tora plant at three different concentrations of 1.0, 2.5, and 5.0?mg/cm² were applied on skin of forearm in man and exposed against adult female mosquitoes. In this observation, this plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes. This is the first report on mosquito adulticidal and repellent activities of the reported C. tora against mosquito vectors from Southern India.     

Chemical composition and larvicidal activity of plant extracts from Clausena dentata (Willd) (Rutaceae) against dengue,malaria, and filariasis vectors

Mosquitoes in the larval stage are attractive targets for pesticides because mosquitoes breed in water, and thus, it is easy to deal with them in this habitat. The use of conventional pesticides in the water sources, however, introduces many risks to people and/or the environment. Natural pesticides, especially those derived from plants, are more promising in this aspect. Aromatic plants and their essential oils are very important sources of many compounds that are used in different respects. Insecticides of botanical origin may serve as suitable alternative to chemical insecticides. Acetone, chloroform, ethyl acetate, methanol, and petroleum benzine leaf extracts of Clausena dentata were tested against the fourth instar larvae of Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti (Diptera: Culicidae). Larval mortality was observed after 24 h of exposure. The highest larval mortality was found in acetone leaf extract, C. quinquefasciatus (LC₅₀?=?0.150278 mg/ml; LC₉₀?=?7.302613 mg/ml), A. aegypti (LC₅₀?=?0.169495 mg/ml; LC₉₀?=?1.10034 mg/ml), and A. stephensi (LC₅₀?=?0.045684 mg/ml; LC90?=?0.045684 mg/ml). GC–MS analysis of plant extracts of acetone solvent revealed 16 compounds, of which the major compounds were benzene,1,2,3-trimethoxy-5-(2-propenyl) (14.97 %), Z,Z-6,28-heptatriactontadien-2-one (6.81 %), 2-allyl-4-methylphenol (28.14 %), 2-allyl-4-methylphenol (17.34 %), and 2,6,10,14,18,22-tetracosahexaene, 2,6,10,15,19,23-hexamethyl (10.35 %). Our result shows acetone leaf extracts of C. dentata have the potential to be used as an ideal eco-friendly approach for mosquito control.