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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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.