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In this study Pergularia daemia unripe fruits were used to synthesize zinc oxide nanoparticles (Pd-ZnONPs). UV-vis Spectroscopy detected the production of ZnONPs. XRD, FTIR, SEM, and TEM studies were used to characterize the synthesized Pd-ZnONPs. Aedes aegypti (Ae. aegypti) third instar larvae were analyzed to diverse concentrations of Pd-unripe fruit extract and Pd-ZnONPs for 24 hours to assess the larvicidal effect. Mortality was also detected in Ae. aegypti larvae under laboratory conditions, with corresponding LC 50 and LC 90 values of 11.11 and 21.20 µg/ml respectively. As a result of this study, the levels of total proteins, esterases, acetylcholine esterase, and phosphatase enzymes in the third instar larvae of Ae. aegypti were significantly lower than the control. These findings suggest that Pd-ZnONPs could be used to suppress mosquito larval populations., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: The use of Hazomalania voyronii, popularly known as hazomalana, to repel mosquitoes and resist against insect attacks is handed down from generation to generation in Madagascar. In the present study, we investigated the ability of the essential oils (EOs) obtained from the stem wood, fresh and dry bark of H. voyronii to keep important mosquito vectors (Aedes aegypti and Culex quinquefasciatus) away, as well as their toxicity on three insect species of agricultural and public health importance (Cx. quinquefasciatus, Musca domestica and Spodoptera littoralis)., Materials and Methods: Hydrodistillation was used to obtain EOs from stem wood, fresh and dry bark. The chemical compositions were achieved by gas chromatography-mass spectrometry (GC-MS). Toxicity assays using stem wood and bark EOs were performed on larvae of Cx. quinquefasciatus and S. littoralis, and adults of M. domestica by WHO and topical application methods, respectively. Mosquito repellent activity of the most effective EO, i.e. the bark one, was determined on human volunteers by arm-in-cage tests, and results were compared with that of the commercial repellent N,N-ddiethyl-m-toluamide (DEET)., Results: The H. voyronii EOs were characterized by oxygenated monoterpenes with perilla aldehyde (30.9-47.9%) and 1,8-cineole (19.7-33.2%) as the main constituents. The fresh and dry bark EOs were the most active on Cx. quinquefasciatus and S. littoralis larvae, respectively, with LC 50 /LD 50 of 65.5  mg L -1 , and 50.5  μg larva -1 ; the EOs from wood and fresh bark displayed the highest toxicity on M. domestica (LD 50 values 60.8 and 65.8 μg adult -1 , respectively). Repellence assay revealed an almost complete protection (>80%) from both mosquito species for 30 min when pure fresh bark EO was applied on the volunteers' arm, while DEET 10% repelled >80% of the mosquitoes up to 120 min from application., Conclusion: The traditional use of the bark EO to repel insects has been demonstrated although an extended-release formulation based on H. voyronii EOs is needed to increase the repellent effect over time. A wide spectrum of insecticidal activity has been provided as well, suggesting a possible use of H. voyronii EOs in the fabrication of green repellents and insecticides useful to control mosquito vectors and agricultural pests., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Mosquitoes act as vectors of devastating pathogens and parasites, representing a key threat for millions of humans and animals worldwide. Eco-friendly control tools are urgently required. We proposed a novel method of fern-mediated biosynthesis of silver nanoparticles (AgNP) using Dicranopteris linearis, acting as a reducing and capping agent. AgNP were characterized by UV-vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), zeta potential and particle size analysis. In mosquitocidal assays, the LC 50 of D. linearis extract against Aedes aegypti ranged from 165.213 (larva I) to 255.055ppm (pupa). LC 50 of D. linearis-synthesized AgNP ranged from 18.905 (larva I) to 29.328ppm (pupa). In the field, the application of D. linearis extract and AgNP (10×LC 50 ) led to 100% larval reduction after 72h. Smoke toxicity experiments conducted against A. aegypti adults showed that D. linearis leaf-, stem- and root-based coils evoked mortality rates comparable to the permethrin-based positive control (58%, 47%, 34%, and 48% respectively). In ovicidal experiments, egg hatchability was reduced by 100% after treatment with 25ppm of AgNP and 300ppm of D. linearis extract. Interestingly, oviposition deterrent assays highlighted that 100ppm of fern extract reduced oviposition rates of more than 65%, while 10ppm of fern-fabricated AgNP reduced oviposition rates of more than 70% in A. aegypti (OAI were -0.52 and -0.55, respectively). Overall, our results highlighted that D. linearis-synthesized AgNP could be useful candidates to develop nano-formulated oviposition deterrents effective against dengue vectors., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Mosquitoes are vectors of devastating pathogens and parasites, causing millions of deaths every year. Dengue is a mosquito-borne viral infection found in tropical and subtropical regions around the world. Recently, dengue transmission has strongly increased in urban and semiurban areas, becoming a major international public health concern. Aedes aegypti (Diptera: Culicidae) is a primary vector of dengue. Shedding light on genetic deviation in A. aegypti populations is of crucial importance to fully understand their molecular ecology and evolution. In this research, haplotype and genetic analyses were conducted using individuals of A. aegypti from 31 localities in the north, southeast, northeast and central regions of Tamil Nadu (South India). The mitochondrial DNA region of cytochrome c oxidase 1 (CO1) gene was used as marker for the analyses. Thirty-one haplotypes sequences were submitted to GenBank and authenticated. The complete haplotype set included 64 haplotypes from various geographical regions clustered into three groups (lineages) separated by three fixed mutational steps, suggesting that the South Indian Ae. aegypti populations were pooled and are linked with West Africa, Columbian and Southeast Asian lineages. The genetic and haplotype diversity was low, indicating reduced gene flow among close populations of the vector, due to geographical barriers such as water bodies. Lastly, the negative values for neutrality tests indicated a bottle-neck effect and supported for low frequency of polymorphism among the haplotypes. Overall, our results add basic knowledge to molecular ecology of the dengue vector A. aegypti, providing the first evidence for multiple introductions of Ae. aegypti populations from Columbia and West Africa in South India.

Mosquito vectors (Diptera: Culicidae) are responsible for transmission of serious diseases worldwide. Mosquito control is being enhanced in many areas, but there are significant challenges, including increasing resistance to insecticides and lack of alternative, cost-effective, and eco-friendly products. To deal with these crucial issues, recent emphasis has been placed on plant materials with mosquitocidal properties. Furthermore, cancers figure among the leading causes of morbidity and mortality worldwide, with approximately 14 million new cases and 8.2 million cancer-related deaths in 2012. It is expected that annual cancer cases will rise from 14 million in 2012 to 22 million within the next two decades. Nanotechnology is a promising field of research and is expected to give major innovation impulses in a variety of industrial sectors. In this study, we synthesized titanium dioxide (TiO2) nanoparticles using the hydrothermal method. Nanoparticles were subjected to different analysis including UV-Vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), zeta potential, and energy-dispersive spectrometric (EDX). The synthesized TiO2 nanoparticles exhibited dose-dependent cytotoxicity against human breast cancer cells (MCF-7) and normal breast epithelial cells (HBL-100). After 24-h incubation, the inhibitory concentrations (IC50) were found to be 60 and 80 μg/mL on MCF-7 and normal HBL-100 cells, respectively. Induction of apoptosis was evidenced by Acridine Orange (AO)/ethidium bromide (EtBr) and 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) staining. In larvicidal and pupicidal experiments conducted against the primary dengue mosquito Aedes aegypti, LC50 values of nanoparticles were 4.02 ppm (larva I), 4.962 ppm (larva II), 5.671 ppm (larva III), 6.485 ppm (larva IV), and 7.527 ppm (pupa). Overall, our results suggested that TiO2 nanoparticles may be considered as a safe tool to build newer and safer mosquitocides and chemotherapeutic agents with little systemic toxicity.

Mosquitoes (Diptera: Culicidae) serve as important vectors for a wide number of parasites and pathogens of huge medical and veterinary importance. Aedes aegypti is a primary dengue vector in tropical and subtropical urban areas. There is an urgent need to develop eco-friendly mosquitocides. In this study, silver nanoparticles (AgNP) were biosynthesized using neem cake, a by-product of the neem oil extraction from the seed kernels of Azadirachta indica. AgNP were characterized using a variety of biophysical methods, including UV-vis spectrophotometry, FTIR, SEM, EDX, and XRD analyses. Furthermore, the neem cake extract and the biosynthesized AgNP were tested for acute toxicity against larvae and pupae of the dengue vector Ae. aegypti. LC50 values achieved by the neem cake extract ranged from 106.53 (larva I) to 235.36 ppm (pupa), while AgNP LC50 ranged from 3.969 (larva I) to 8.308 ppm (pupa). In standard laboratory conditions, the predation efficiency of a Carassius auratus per day was 7.9 (larva II) and 5.5 individuals (larva III). Post-treatment with sub-lethal doses of AgNP, the predation efficiency was boosted to 9.2 (larva II) and 8.1 individuals (larva III). The genotoxic effect of AgNP was studied on C. auratus using the comet assay and micronucleus frequency test. DNA damage was evaluated on peripheral erythrocytes sampled at different time intervals from the treatment; experiments showed no significant damages at doses below 12 ppm. Overall, this research pointed out that neem cake-fabricated AgNP are easy to produce, stable over time, and can be employed at low dosages to reduce populations of dengue vectors, with moderate detrimental effects on non-target mosquito natural enemies.

Dengue is a mosquito-borne viral disease that has rapidly spread in all regions of the world in recent years. Female mosquitoes, mainly Aedes aegypti, transmit dengue. Approximately 3,900 million people, in 128 countries, are at risk of dengue infection. Recently, a focus has been provided on the potential of green-synthesized nanoparticles as inhibitors of the production of dengue viral envelope (E) protein in Vero cells and downregulators of the expression of dengue viral E gene. Algae are an outstanding reservoir of novel compounds, which may help in the fight against mosquito-borne diseases. In this research, silver nanoparticles (AgNP) were rapidly synthesized using a cheap extract of the alga Centroceras clavulatum. AgNP were characterized by UV–vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray diffraction (XRD). In mosquitocidal assays, LC50 values of C. clavulatum extract against A. aegypti larvae and pupae were 269.361 ppm (larva I), 309.698 ppm (larva II), 348.325 ppm (larva III), 387.637 ppm (larva IV), and 446.262 ppm (pupa). C. clavulatum extract also exhibited moderate antioxidant activity, both in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging assays. LC50 values of C. clavulatum-synthesized AgNP were 21.460 ppm (larva I), 23.579 ppm (larva II), 25.912 ppm (larva III), 29.155 ppm (larva IV), and 33.877 ppm (pupa). Furthermore, C. clavulatum-synthesized AgNP inhibited dengue (serotype dengue virus type-2 (DEN-2)) viral replication in Vero cells. Notably, 50 μg/ml of green-synthesized AgNP showed no cytotoxicity on Vero cells while reduced DEN-2 viral growth of more than 80%; 12.5 μg/ml inhibited viral growth of more than 50%. Cellular internalization assays highlighted that untreated infected cells showed high intensity of fluorescence emission, which denotes high level of viral internalization. Conversely, AgNP-treated infected cells showed reduced levels of fluorescence, failing to show significant viral load. Overall, our study showed that alga-mediated synthesis of metal nanoparticles may be considered to develop newer, safer, and cheap tools in the fight against the dengue virus, serotype DEN-2, and its vector A. aegypti, with little cytotoxicity on mammalian cells.

Aedes albopictus is an important arbovirus vector, including dengue. Currently, there is no specific treatment for dengue. Its prevention solely depends on effective vector control measures. In this study, silver nanoparticles (AgNPs) were biosynthesized using a cheap leaf extract of Berberis tinctoria as reducing and stabilizing agent and tested against Ae. albopictus and two mosquito natural enemies. AgNPs were characterized by using UV–vis spectrophotometry, X-ray diffraction, and scanning electron microscopy. In laboratory conditions, the toxicity of AgNPs was evaluated on larvae and pupae of Ae. albopictus. Suitability Index/Predator Safety Factor was assessed on Toxorhynchites splendens and Mesocyclops thermocyclopoides. The leaf extract of B. tinctoria was toxic against larval instars (I–IV) and pupae of Ae. albopictus; LC50 was 182.72 ppm (I instar), 230.99 ppm (II), 269.65 ppm (III), 321.75 ppm (IV), and 359.71 ppm (pupa). B. tinctoria-synthesized AgNPs were highly effective, with LC50 of 4.97 ppm (I instar), 5.97 ppm (II), 7.60 ppm (III), 9.65 ppm (IV), and 14.87 ppm (pupa). Both the leaf extract and AgNPs showed reduced toxicity against the mosquito natural enemies M. thermocyclopoides and T. splendens. Overall, this study firstly shed light on effectiveness of B. tinctoria-synthesized AgNPs as an eco-friendly nanopesticide, highlighting the concrete possibility to employ this newer and safer tool in arbovirus vector control programs.

Aedes aegypti is a primary vector of dengue, a mosquito-borne viral disease infecting 50-100 million people every year. Here, we biosynthesised mosquitocidal silver nanoparticles (AgNP) using the aqueous leaf extract of Crotalaria verrucosa. The green synthesis of AgNP was studied by UV-vis spectroscopy, SEM, EDX and FTIR. C. verrucosa-synthesised AgNPs were toxic against A. aegypti larvae and pupae. LC50 of AgNP ranged from 3.496 ppm (I instar larvae) to 17.700 ppm (pupae). Furthermore, we evaluated the predatory efficiency of dragonfly nymphs, Brachydiplax sobrina, against II and III instar larvae of A. aegypti in an aquatic environment contaminated with ultra-low doses of AgNP. Under standard laboratory conditions, predation after 24 h was 87.5% (II) and 54.7% (III). In an AgNP-contaminated environment, predation was 91 and 75.5%, respectively. Overall, C. verrucosa-synthesised AgNP could be employed at ultra-low doses to reduce larval population of dengue vectors enhancing predation rates of dragonfly nymphs.

Introduction: Development of new and improved mosquito control methods, that are economically sustainable and effective, is a critical requirement in the management of vector-borne epidemic diseases. Aedes albopictus is one of the main vectors of various important pathogens in the tropics, which now have the potential to also spread in temperate regions, owing to the environmental and climate changes in act., Materials and Methods: We report about the isolation of steroidal saponins from Dracaena arborea by fractionation followed by column separation. The obtained fractions and/or pure compounds were tested by biological essays for their insecticidal activity against A. albopictus larvae., Results: Various compounds were found to exert larvicidal effects. In specific, spiroconazole A demonstrated the best insecticidal activity, showing LT50 value of 57.23 hours at 25 ppm., Discussion: We finally discuss about the value of this finding in the context of the present strategies of Integrated Mosquito Management.

Mosquito-borne diseases represent a deadly threat for millions of people worldwide. However, the use of synthetic insecticides to control Culicidae may lead to high operational costs and adverse non-target effects. Plant-borne compounds have been proposed for rapid extracellular synthesis of mosquitocidal nanoparticles. Their impact against biological control agents of mosquito larval populations has been poorly studied. We synthesized silver nanoparticles (AgNP) using the aqueous leaf extract of Mimusops elengi as a reducing and stabilizing agent. The formation of AgNP was studied using different biophysical methods, including UV-vis spectrophotometry, TEM, XRD, EDX and FTIR. Low doses of AgNP showed larvicidal and pupicidal toxicity against the malaria vector Anopheles stephensi and the arbovirus vector Aedes albopictus. AgNP LC50 against A. stephensi ranged from 12.53 (I instar larvae) to 23.55 ppm (pupae); LC50 against A. albopictus ranged from 11.72 ppm (I) to 21.46 ppm (pupae). In the field, the application of M. elengi extract and AgNP (10 × LC50) led to 100 % larval reduction after 72 h. In adulticidal experiments, AgNP showed LC50 of 13.7 ppm for A. stephensi and 14.7 ppm for A. albopictus. The predation efficiency of Gambusia affinis against A. stephensi and A. albopictus III instar larvae was 86.2 and 81.7 %, respectively. In AgNP-contaminated environments, predation was 93.7 and 88.6 %, respectively. This research demonstrates that M. elengi-synthesized AgNP may be employed at ultra-low doses to reduce larval populations of malaria and arbovirus vectors, without detrimental effects on predation rates of mosquito natural enemies, such as larvivorous fishes.

Two of the most important challenges facing humanity in the 21st century comprise food production and disease control. Eco-friendly control tools against mosquito vectors and agricultural pests are urgently needed. Insecticidal products of marine origin have a huge potential to control these pests. In this research, we reported a single-step method to synthesize silver nanoparticles (AgNP) using the aqueous leaf extract of the seaweed Hypnea musciformis, a cheap, nontoxic and eco-friendly material, that worked as reducing and stabilizing agent during the biosynthesis. The formation of AgNP was confirmed by surface plasmon resonance band illustrated in UV-vis spectrophotometer. AgNP were characterized by FTIR, SEM, EDX and XRD analyses. AgNP were mostly spherical in shape, crystalline in nature, with face-centered cubic geometry, and their mean size was 40-65nm. Low doses of H. musciformis aqueous extract and seaweed-synthesized AgNP showed larvicidal and pupicidal toxicity against the dengue vector Aedes aegypti and the cabbage pest Plutella xylostella. The LC50 value of AgNP ranged from 18.14 to 38.23ppm for 1st instar larvae (L1) and pupae of A. aegypti, and from 24.5 to 38.23ppm for L1 and pupae of P. xylostella. Both H. musciformis extract and AgNP strongly reduced longevity and fecundity of A. aegypti and P. xylostella adults. This study adds knowledge on the toxicity of seaweed borne insecticides and green-synthesized AgNP against arthropods of medical and agricultural importance, allowing us to propose the tested products as effective candidates to develop newer and cheap pest control tools., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Aedes aegypti is a primary vector of dengue and chikungunya. The use of synthetic insecticides to control Aedes populations often leads to high operational costs and adverse non-target effects. Botanical extracts have been proposed for rapid extracellular synthesis of mosquitocidal nanoparticles, but their impact against predators of mosquito larvae has not been well studied. We propose a single-step method for the biosynthesis of silver nanoparticles (AgNP) using the extract of Artemisia vulgaris leaves as a reducing and stabilizing agent. AgNP were characterized by UV-vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray diffraction (XRD). SEM and XRD showed that AgNP were polydispersed, crystalline, irregularly shaped, with a mean size of 30-70 nm. EDX confirmed the presence of elemental silver. FTIR highlighted that the functional groups from plant metabolites capped AgNP, stabilizing them over time. We investigated the mosquitocidal properties of A. vulgaris leaf extract and green-synthesized AgNP against larvae and pupae of Ae. aegypti. We also evaluated the predatory efficiency of Asian bullfrog tadpoles, Hoplobatrachus tigerinus, against larvae of Ae. aegypti, under laboratory conditions and in an aquatic environment treated with ultra-low doses of AgNP. AgNP were highly toxic to Ae. aegypti larval instars (I-IV) and pupae, with LC50 ranging from 4.4 (I) to 13.1 ppm (pupae). In the lab, the mean number of prey consumed per tadpole per day was 29.0 (I), 26.0 (II), 21.4 (III), and 16.7 (IV). After treatment with AgNP, the mean number of mosquito prey per tadpole per day increased to 34.2 (I), 32.4 (II), 27.4 (III), and 22.6 (IV). Overall, this study highlights the importance of a synergistic approach based on biocontrol agents and botanical nano-insecticides for mosquito control.

Dengue is an arthropod-borne viral infection mainly vectored through the bite of Aedes mosquitoes. Recently, its transmission has strongly increased in urban and semi-urban areas of tropical and sub-tropical regions worldwide, becoming a major international public health concern. There is no specific treatment for dengue. Its prevention and control solely depends on effective vector control measures. In this study, we proposed the green-synthesis of silver nanoparticles (AgNP) as a novel and effective tool against the dengue serotype DEN-2 and its major vector Aedes aegypti. AgNP were synthesized using the Moringa oleifera seed extract as reducing and stabilizing agent. AgNP were characterized using a variety of biophysical methods including UV-vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and sorted for size categories. AgNP showed in vitro antiviral activity against DEN-2 infecting vero cells. Viral titer was 7 log10 TCID50/ml in control (AgNP-free), while it dropped to 3.2 log10 TCID50/ml after a single treatment with 20 μl/ml of AgNP. After 6 h, DEN-2 yield was 5.8 log10 PFU/ml in the control, while it was 1.4 log10 PFU/ml post-treatment with AgNP (20 μl/ml). AgNP were highly effective against the dengue vector A. aegypti, with LC50 values ranging from 10.24 ppm (I instar larvae) to 21.17 ppm (pupae). Overall, this research highlighted the concrete potential of green-synthesized AgNP in the fight against dengue and its primary vector A. aegypti. Further research on structure-activity relationships of AgNP against other dengue serotypes is urgently required.

Plant-borne compounds can be employed to synthesize mosquitocidal nanoparticles that are effective at low doses. However, how they affect the activity of mosquito predators in the aquatic environment is unknown. In this study, we synthesized gold nanoparticles (AuN) using the leaf extract of Cymbopogon citratus, which acted as a reducing and capping agent. AuN were characterized by a variety of biophysical methods and sorted for size in order to confirm structural integrity. C. citratus extract and biosynthesized AuN were tested against larvae and pupae of the malaria vector Anopheles stephensi and the dengue vector Aedes aegypti. LC₅₀ of C. citratus extract ranged from 219.32 ppm to 471.36 ppm. LC₅₀ of AuN ranged from 18.80 ppm to 41.52 ppm. In laboratory, the predatory efficiency of the cyclopoid crustacean Mesocyclops aspericornis against A. stephensi larvae was 26.8% (larva I) and 17% (larva II), while against A. aegypti was 56% (I) and 35.1% (II). Predation against late-instar larvae was minimal. In AuN-contaminated environment,predation efficiency against A. stephensi was 45.6% (I) and 26.7% (II), while against A. aegypti was 77.3% (I) and 51.6% (II). Overall, low doses of AuN may help to boost the control of Anopheles and Aedes larval populations in copepod-based control programs., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Mosquitoes are vectors of devastating pathogens and parasites, causing millions of deaths every year. Dengue is a mosquito-borne viral infection found in tropical and subtropical regions around the world. Recently, transmission has strongly increased in urban and semiurban areas, becoming a major international public health concern. Aedes aegypti (Diptera: Culicidae) is the primary vector of dengue. The use of synthetic insecticides to control Aedes mosquitoes lead to high operational costs and adverse nontarget effects. In this scenario, eco-friendly control tools are a priority. We proposed a novel method to synthesize silver nanoparticles using the aqueous leaf extract of Phyllanthus niruri, a cheap and nontoxic material. The UV-vis spectrum of the aqueous medium containing silver nanostructures showed a peak at 420 nm corresponding to the surface plasmon resonance band of nanoparticles. SEM analyses of the synthesized nanoparticles showed a mean size of 30-60 nm. EDX spectrum showed the chemical composition of the synthesized nanoparticles. XRD highlighted that the nanoparticles are crystalline in nature with face-centered cubic geometry. Fourier transform infrared spectroscopy (FTIR) of nanoparticles exhibited prominent peaks 3,327.63, 2,125.87, 1,637.89, 644.35, 597.41, and 554.63 cm(-1). In laboratory assays, the aqueous extract of P. niruri was toxic against larval instars (I-IV) and pupae of A. aegypti. LC50 was 158.24 ppm (I), 183.20 ppm (II), 210.53 ppm (III), 210.53 ppm (IV), and 358.08 ppm (pupae). P. niruri-synthesized nanoparticles were highly effective against A. aegypti, with LC50 of 3.90 ppm (I), 5.01 ppm (II), 6.2 ppm (III), 8.9 ppm (IV), and 13.04 ppm (pupae). In the field, the application of silver nanoparticles (10 × LC50) lead to A. aegypti larval reduction of 47.6%, 76.7% and 100%, after 24, 48, and 72 h, while the P. niruri extract lead to 39.9%, 69.2 % and 100 % of reduction, respectively. In adulticidal experiments, P. niruri extract and nanoparticles showed LC50 and LC90 of 174.14 and 6.68 ppm and 422.29 and 23.58 ppm, respectively. Overall, this study highlights that the possibility to employ P. niruri leaf extract and green-synthesized silver nanoparticles in mosquito control programs is concrete, since both are effective at lower doses if compared to synthetic products currently marketed, thus they could be an advantageous alternative to build newer and safer tools against dengue vectors.

Neem seed oil (NSO) of Azadirachta indica (Meliaceae) contains more than 100 determined biologically active compounds, and many formulations deriving from them showed toxicity, antifeedancy and repellence against a number of arthropod pests. However, it is widely known that botanical products can differ in their chemical composition and bioactivity, as function of the production site and production process. We used high-performance thin layer chromatography (HPTLC) to investigate differences in chemical constituents of NSOs from three production sites. HPTLC analyses showed several differences in chemical abundance and diversity among NSOs, with special reference to limonoids. Furthermore, the three NSOs and their fractions of increasing polarities [i.e. ethyl acetate (EA) fraction and butanol (BU) fraction] were evaluated for larvicidal toxicity and field oviposition deterrence against the Asian tiger mosquito, Aedes albopictus, currently the most invasive mosquito worldwide. Results from bioactivity experiments showed good toxicity of NSOs and EA fractions against A. albopictus fourth instar larvae (with LC50 values ranging from 142.28 to 209.73 ppm), while little toxicity was exerted by BU fractions. A significant effect of the production site and dosage was also found and is probably linked to differences in abundance of constituents among samples, as highlighted by HPTLC analyses. NSOs and EAs were also able to deter A. albopictus oviposition in the field (effective repellence values ranging from 98.55 to 70.10%), while little effectiveness of BU fractions was found. Concerning ovideterrent activity, no difference due to the production site was found. This is the first report concerning larvicidal toxicity of NSO against A. albopictus and ovideterrence against Culicidae in the field. The chance to use chemicals from the NSO EA fraction seems promising, since they are effective at lower doses, if compared to synthetic products currently marketed, and could be an advantageous alternative to build newer and safer mosquito control tools.

Industrial plant-borne by-products can be sources of low-cost chemicals, potentially useful to build eco-friendly control strategies against mosquitoes. Neem cake is a cheap by-product of neem oil extraction obtained by pressing the seeds of Azadirachta indica. Neem products are widely used as insecticides since rarely induce resistance because their multiple mode of action against insect pests and low-toxicity rates have been detected against vertebrates. In this research, we used field bioassays to assess the effective oviposition repellence of neem cake fractions of increasing polarity [n-hexane (A), methanol (B), ethyl acetate (C), n-butanol (D), and aqueous (E) fraction] against Aedes albopictus, currently the most invasive mosquito worldwide. These fractions, already characterized for low nortriterpenoids contents by HPLC analyses, were analyzed for their total content by HPTLC, highlighting striking differences in their chemical composition. Field results showed that B, A, and C tested at 100 ppm exerted higher effective repellence over the control (71.33, 88.59, and 73.49% of ER, respectively), while E and D did not significantly deter A. albopictus oviposition (17.06 and 22.72% of ER, respectively). The highest oviposition activity index was achieved by A (-0.82), followed by C (-0.63), and B (-0.62). Lower OAIs were achieved by D (-0.14) and E (-0.09). On the basis of our results, we believe that A, B, and C are very promising as oviposition deterrents against the arbovirus vector A. albopictus since they are proved as rich in active metabolites, cheap, and really effective at low doses.

New pesticides based on natural products are urgently needed, in consideration of their environmental care and lower collateral effects. Neem oil, the main product obtained from Azadiractha indica A. Juss, commonly known as neem tree, is mainly used in medical devices, cosmetics and soaps, as well as important insecticide. Manufacturing of neem oil first includes the collection of the neem seeds as raw material used for the extraction. Neem cake is the waste by-product remaining after extraction processes. The quality of the oil, as that of the cake, strictly depends from the quality of seeds as well as from the type of extraction processes used, which strongly influences the chemical composition of the product. Currently, the different types of commercial neem cake on the market are roughly identified as oiled and deoiled cake, but several other differences can be detected. The differences are relevant and must be determined, to obtain the necessary correlation between chemical constitution and larvicidal activities. Six different batches of neem cake, marketed by several Indian and European companies, were analyzed by HPLC and HPTLC, and their fingerprints compared, obtaining information about the different compositions, focusing in particular on nortriterpenes, considered as the main active components of neem oil. Therefore, the chemical composition of each cake was connected with the biological activitiy, i.e., the effects of the extracts of the six neem cakes were tested on eggs and larvae of Aedes albopictus (Stegomyia albopicta) (Diptera: Culicidae), commonly known as Asian tiger mosquito. The results confirmed the previously reported larvicide effects of neem cake that, however, can now be related to the chemical composition, in particular with nortriterpenes, allowing in that way to discriminate between the quality of the various marketed products, as potential domestic insecticides.

In order to investigate its insecticide potential, the neem cake methanol extract was first analyzed and then separated by different solvents. The high-performance liquid chromatography analysis showed that the neem cake methanol extract still contained relevant quantities of nortriterpenes. Fractions of increasing polarity were obtained from the separation process: hexane fraction (Hp), EtOAc fraction (Ep), n-BuOH fraction (Bp), and aqueous fraction (Wp). The activity of the fractions on Aedes albopictus (Skuse) eggs and larvae was tested, and the Ep fraction exhibits the most relevant larvicide effect. The nuclear magnetic resonance fingerprint analysis of this phytocomplex isolated on EtOAc fraction was performed. The larvicidal effectiveness of the phytocomplex isolated on EtOAc, compared to that of pure and commercial azadirachtin solutions of different concentrations, was checked. The results showed that the activity of the phytocomplex, as a whole, was significantly higher than those of isolated compound solutions. As a consequence, the neem cake is a promising low-cost, easily available on the market, and natural resource to develop a new bioinsecticide, mainly in developing countries.