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.

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.

Mosquitoes represent an important threat for lives of millions of people worldwide, acting as vectors for devastating pathogens, such as malaria, yellow fever, dengue, and West Nile. In addition, pathogens and parasites polluting water also constitute a severe plague for populations of developing countries. Here, we investigated the mosquitocidal and antibacterial properties of Aloe vera leaf extract and silver nanoparticles synthesized using A. vera extract. Mosquitocidal properties were assessed in laboratory against larvae (I-IV instar) and pupae of the malaria vector Anopheles stephensi. Green-synthesized silver nanoparticles were tested against An. stephensi also in field conditions. Antibacterial properties of nanoparticles were evaluated against Bacillus subtilis, Klebsiella pneumoniae, and Salmonella typhi using the agar disk diffusion and minimum inhibitory concentration protocol. The synthesized silver nanoparticles were characterized by UV-vis spectrum, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and X-ray diffraction (XRD). In laboratory conditions, the A. vera extract was toxic against An. stephensi larvae and pupae, even at low dosages. LC50 were 48.79 ppm (I instar), 59.09 ppm (II instar), 70.88 ppm (III instar), 83.58 ppm (IV instar), and 152.55 ppm (pupae). Green-synthesized silver nanoparticles were highly toxic against An. stephensi. LC50 were 3.825 ppm (I instar), 4.119 ppm (II instar), 4.982 ppm (III instar), 5.711 ppm (IV instar), and 6.113 ppm (pupae). In field conditions, the application of A. vera-synthesized silver nanoparticles (10 × LC50) leads to An. stephensi larval reduction of 74.5, 86.6, and 97.7%, after 24, 48, and 72 h, respectively. Nanoparticles also showed antibacterial properties, and the maximum concentration tested (150 mg/L) evoked an inhibition zone wider than 80 mm in all tested bacterium species. This study adds knowledge about the use of green synthesis of nanoparticles in medical entomology and parasitology, allowing us to propose A. vera-synthesized silver nanoparticles as effective candidates to develop newer and safer mosquitocidal control tools.

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Nearly 1.4 billion people in 73 countries worldwide are threatened by lymphatic filariasis, a parasitic infection that leads to a disease commonly known as elephantiasis. Filariasis is vectored by mosquitoes, with special reference to the genus Culex. The main control tool against mosquito larvae is represented by treatments with organophosphates and insect growth regulators, with negative effects on human health and the environment. Recently, green-synthesized nanoparticles have been proposed as highly effective larvicidals against mosquito vectors. In this research, we attempted a reply to the following question: do green-synthesized nanoparticles affect predation rates of copepods against mosquito larvae? We proposed a novel method of seaweed-mediated synthesis of silver nanoparticles using the frond extract of Caulerpa scalpelliformis. The toxicity of the seaweed extract and silver nanoparticles was assessed against the filarial vector Culex quinquefasciatus. Then, we evaluated the predatory efficiency of the cyclopoid crustacean Mesocyclops longisetus against larval instars of C. quinquefasciatus in a nanoparticle-contaminated water environment. Green-synthesized silver nanoparticles were characterized by UV-vis spectrum, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and X-ray diffraction (XRD). In mosquitocidal assays, the LC₅₀ values of the C. scalpelliformis extract against C. quinquefasciatus were 31.38 ppm (I), 46.49 ppm (II), 75.79 ppm (III), 102.26 ppm (IV), and 138.89 ppm (pupa), while LC₅₀ of silver nanoparticles were 3.08 ppm, (I), 3.49 ppm (II), 4.64 ppm (III), 5.86 ppm (IV), and 7.33 ppm (pupa). The predatory efficiency of the copepod M. longisetus in the control treatment was 78 and 59% against I and II instar larvae of C. quinquefasciatus. In a nanoparticle-contaminated environment, predation efficiency was 84 and 63%, respectively. Predation was higher against first instar larvae over other instars. Overall, our study showed that seaweed-synthesized silver nanoparticles can be proposed in synergy with biological control agents against Culex larvae, since their use leads to little detrimental effects against aquatic predators, such as copepods.

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The development of parasites and pathogens resistant to synthetic drugs highlighted the needing of novel, eco-friendly and effective control approaches. Recently, metal nanoparticles have been proposed as highly effective tools towards cancer cells and Plasmodium parasites. In this study, we synthesized silver nanoparticles (EW-AgNP) using Eudrilus eugeniae earthworms as reducing and stabilizing agents. EW-AgNP showed plasmon resonance reduction in UV-vis spectrophotometry, the functional groups involved in the reduction were studied by FTIR spectroscopy, while particle size and shape was analyzed by FESEM. The effect of EW-AgNP on in vitro HepG2 cell proliferation was measured using MTT assays. Apoptosis assessed by flow cytometry showed diminished endurance of HepG2 cells and cytotoxicity in a dose-dependent manner. EW-AgNP were toxic to Anopheles stephensi larvae and pupae, LC(50) were 4.8 ppm (I), 5.8 ppm (II), 6.9 ppm (III), 8.5 ppm (IV), and 15.5 ppm (pupae). The antiplasmodial activity of EW-AgNP was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of Plasmodium falciparum. EW-AgNP IC(50) were 49.3 μg/ml (CQ-s) and 55.5 μg/ml (CQ-r), while chloroquine IC(50) were 81.5 μg/ml (CQ-s) and 86.5 μg/ml (CQ-r). EW-AgNP showed a valuable antibiotic potential against important pathogenic bacteria and fungi. Concerning non-target effects of EW-AgNP against mosquito natural enemies, the predation efficiency of the mosquitofish Gambusia affinis towards the II and II instar larvae of A. stephensi was 68.50% (II) and 47.00% (III), respectively. In EW-AgNP-contaminated environments, predation was boosted to 89.25% (II) and 70.75% (III), respectively. Overall, this research highlighted the EW-AgNP potential against hepatocellular carcinoma, Plasmodium parasites and mosquito vectors, with little detrimental effects on mosquito natural enemies.

Each year, mosquito-borne diseases infect nearly 700 million people, resulting more than one million deaths. In this study, we proposed a Metarhizium anisopliae-based method of green synthesis of silver nanoparticles to control the rural malaria vector Anopheles culicifacies. Silver nanoparticles were characterized by UV–Vis spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, energy-dispersive X-ray analysis, and X-ray diffraction. In acute toxicity experiments, larvae (I–IV instar) and pupae of A. culicifacies were exposed to M. anisopliae-synthesized silver nanoparticles (15, 30, 45, 60, and 75 ppm). LC50 of silver nanoparticles was 32.8 ppm (I), 39.8 ppm (II), 45.9 ppm (III), 51.9 (IV), and 60.0 ppm (pupa). Lower dosages of myco-synthesized silver nanoparticles have detrimental effects on larval and pupal development of A. culicifacies. EI50 was 14.9 ppm. Overall, this research highlighted that myco-synthesized silver nanoparticles can be proposed as effective tools for eco-friendly control of the rural malaria vector A. culicifacies