Mosquitoes are responsible for the transmission of many pathogens and parasites, which cause serious diseases in humans and animals. Currently, botanical products have been suggested as alternative tools in the fight against arthropod vectors. In this study, the essential oil (EO) extracted from Zingiber cernuum was tested as larvicide and oviposition deterrent on six mosquito species of public health relevance, including malaria and Zika virus vectors. The EO showed high toxicity on third instar larvae of Anopheles stephensi (LC₅₀ = 41.34 μg/ml), Aedes aegypti (LC₅₀ = 44.88 μg/ml), Culex quinquefasciatus (LC₅₀ = 48.44 μg/ml), Anopheles subpictus (LC₅₀ = 51.42 μg/ml), Aedes albopictus (LC₅₀ = 55.84 μg/ml), and Culex tritaeniorhynchus (LC₅₀ = 60.20 μg/ml). In addition, low doses of Z. cernuum EO reduced oviposition rates in six mosquito species. The acute toxicity of Z. cernuum EO on four mosquito predators was scarce; LC₅₀ ranged from 3119 to 11,233 μg/ml. Overall, our results revealed that the Z. cernuum EO can be considered for the development of effective and environmental-friendly mosquito larvicides and oviposition deterrents.

The use of synthetic pesticides to control vector populations is detrimental to human health and the environment and may lead to the development of resistant strains. Plants can be alternative sources of safer compounds effective on mosquito vectors. In this study, the mosquito larvicidal activity of Boswellia ovalifoliolata leaf essential oil (EO) was evaluated against Anopheles stephensi, Anopheles subpictus, Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Culex tritaeniorhynchus. GC-MS revealed that the B. ovalifoliolata EO contained at least 20 compounds. The main constituents were β-pinene, α-terpineol, and caryophyllene. In acute toxicity assays, the EO was toxic to larvae of An. stephensi, Ae. aegypti, Cx. quinquefasciatus, An. subpictus, Ae. albopictus, and Cx. tritaeniorhynchus with LC₅₀ values of 61.84, 66.24, 72.47, 82.26, 89.80, and 97.95 μg/ml, respectively. B. ovalifoliolata EO was scarcely toxic to mosquito fishes, backswimmers, and water bugs predating mosquito larvae with LC₅₀ from 4186 to 14,783 μg/ml. Overall, these results contribute to develop effective and affordable instruments to magnify the reliability of Culicidae control programs.

Anopheles stephensi acts as vector of Plasmodium parasites, which are responsible for malaria in tropical and subtropical areas worldwide. Currently, malaria management is a big challenge due to the presence of insecticide-resistant strains as well as to the development of Plasmodium species highly resistant to major antimalarial drugs. Therefore, the present study focused on biosurfactant produced by two bacteria Bacillus subtilis A1 and Pseudomonas stutzeri NA3, evaluating them for insecticidal applications against malaria mosquitoes. The produced biosurfactants were characterized using FT-IR spectroscopy and gas chromatography-mass spectrometry (GC-MS), which confirmed that biosurfactants had a lipopeptidic nature. Both biosurfactants were tested against larvae and pupae of A. stephensi. LC₅₀ values were 3.58 (larva I), 4.92 (II), 5.73 (III), 7.10 (IV), and 7.99 (pupae) and 2.61 (I), 3.68 (II), 4.48 (III), 5.55 (IV), and 6.99 (pupa) for biosurfactants produced by B. subtilis A1 and P. stutzeri NA3, respectively. Treatments with bacterial surfactants led to various physiological changes including longer pupal duration, shorter adult oviposition period, and reduced longevity and fecundity. To the best of our knowledge, there are really limited reports on the mosquitocidal and physiological effects due to biosurfactant produced by bacterial strains. Overall, the toxic activity of these biosurfactant on all young instars of A. stephensi, as well as their major impact on adult longevity and fecundity, allows their further consideration for the development of insecticides in the fight against malaria mosquitoes.

The development of novel mosquito control tools is a key prerequisite to build effective and reliable Integrated Vector Management strategies. Here, we proposed a novel method using cigarette butts for the synthesis of Ag nanostructures toxic to young instars of the malaria vector Anopheles stephensi, chloroquine (CQ)-resistant malaria parasites Plasmodium falciparum and microbial pathogens. The non-target impact of these nanomaterials in the aquatic environment was evaluated testing them at sub-lethal doses on the predatory copepod Mesocyclops aspericornis. Cigarette butt-synthesized Ag nanostructures were characterized by UV–vis and FTIR spectroscopy, as well as by EDX, SEM and XRD analyses. Low doses of cigarette butt extracts (with and without tobacco) showed larvicidal and pupicidal toxicity on An. stephensi. The LC₅₀ of cigarette butt-synthesized Ag nanostructures ranged from 4.505 ppm (I instar larvae) to 8.070 ppm (pupae) using smoked cigarette butts with tobacco, and from 3.571 (I instar larvae) to 6.143 ppm (pupae) using unsmoked cigarette butts without tobacco. Smoke toxicity experiments conducted against adults showed that unsmoked cigarette butts-based coils led to mortality comparable to permethrin-based positive control (84.2 and 91.2%, respectively). A single treatment with cigarette butts extracts and Ag nanostructures significantly reduced egg hatchability of An. stephensi. Furthermore, the antiplasmodial activity of cigarette butt extracts (with and without tobacco) and synthesized Ag nanostructures was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of P. falciparum. The lowest IC₅₀ values were achieved by cigarette butt extracts without tobacco, they were 54.63 μg/ml (CQ-s) and 63.26 μg/ml (CQ-r); while Ag nanostructure IC₅₀ values were 72.13 μg/ml (CQ-s) and 77.33 μg/ml (CQ-r). In MIC assays, low doses of the Ag nanostructures inhibited the growth of Bacillus subtilis, Klebsiella pneumoniae and Salmonella typhi. Finally, the predation efficiency of copepod M. aspericornis towards larvae of An. stephensi did not decrease in a nanoparticle-contaminated environment, if compared to control predation assays. Overall, the present research would suggest that an abundant hazardous waste, such as cigarette butts, can be turned to an important resource for nanosynthesis of highly effective antiplasmodials and insecticides.

Microbial control agents offer alternatives to chemical pest control, as they can be more selective than chemical insecticides. The present study evaluates the mosquito larvicidal and pupicidal potential of fungus mycelia using ethyl acetate and methanol solvent extracts produced by Aspergillus terreus against Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti. The A. terreus mycelia were extracted after 15 days from Sabouraud dextrose broth medium. The ethyl acetate extracts showed lethal concentration that kills 50 % of the exposed larvae (LC₅₀) and lethal concentration that kills 90 % of the exposed larvae (LC₉₀) values of the first, second, third, and fourth instar larvae of An. stephensi (LC₅₀ = 97.410, 102.551, 29.802, and 8.907; LC₉₀ = 767.957, 552.546, 535.474, and 195.677 μg/ml), Cx. quinquefasciatus (LC₅₀ = 89.584, 74.689, 68.265, and 67.40; LC₉₀ = 449.091, 337.355, 518.793, and 237.347 μg/ml), and Ae. aegypti (LC₅₀ = 83.541, 84.418, 80.407, and 95.926; LC₉₀ = 515.464, 443.167, 387.910, and 473.998 μg/ml). Pupicidal activity of mycelium extracts was tested against An. stephensi (LC₅₀ = 25.228, LC₉₀ = 140.487), Cx. quinquefasciatus (LC₅₀ = 54.525, LC₉₀ = 145.366), and Ae. aegypti (LC₅₀ = 10.536, LC₉₀ = 63.762 μg/ml). At higher concentration (500 μg/ml), mortality starts within the first 6 h of exposure. One hundred percent mortality occurs at 24-h exposure. The overall result observed that effective activity against selected mosquito larvae and pupae after 24 h was a dose and time-dependent activity. These ensure that the resultant mosquito population reduction is substantial even where the larvicidal and pupicidal potential is minimal. The FTIR spectra of ethyl acetate extract reflect prominent peaks (3448.32, 3000.36, 2914.59, 2118.73, 1668.21, 1436.87, 1409.02, 954.33, 901.13, and 704.67 cm⁻¹). The spectra showed a sharp absorption band at 1314.66 cm⁻¹ assigned to wagging vibration of the C–H group. The band at 1023.59 cm⁻¹ developed for C–O and C=N, respectively, and was commonly found in carboxylic acid and amine groups. GC–MS analysis of ethyl acetate extracts showed the presence of six compounds, of which the major compounds were identified as n-hexadecanoic acid (15.31 %) and methyl 12,15-octadecadienoate (31.989 %), based on their peak molecular weight. The HPLC analysis result highlights that the A. terreus ethyl acetate extract was compared with pure n-hexadecanoic acid which resulted in similar retention time of 19.52 and 19.38, respectively. Thus, the active compound produced by this species would be more useful against vectors responsible for diseases of public health importance. This is the first report on mosquito larvicidal and pupicidal activity of ethyl acetate extract produced by A. terreus species.

Mosquitoes are principal vector of several vector-borne diseases affecting human beings leading to thousands of deaths per year and responsible for transmitting diseases like malaria, dengue, chikungunya, yellow fever, Zika virus, Japanese encephalitis, and lymphatic filariasis. In the present study, we evaluated the different solvent extracts of mangrove Avicennia marina for their toxicity against larvae of three major mosquito vectors, as well as selected microbial pathogens. The larvicidal mortality of third instars was observed after 24 h. Highest larval mortality was found for the acetone extract of A. marina against Culex quinquefasciatus (LC₅₀ = 0.197 mg/ml; LC₉₀ = 1.5011 mg/ml), Anopheles stephensi (LC₅₀ = 0.176 mg/ml; LC₉₀ = 3.6290 mg/ml), and Aedes aegypti (LC₅₀ = 0.164 mg/ml; LC₉₀ = 4.3554 mg/ml). GC-MS analysis of acetone extract revealed 5 peaks, i.e., 1-hexyl-2-nitrocyclohexane (3.229%), eicosanoic acid (40.582%), cis-9-hexadecenal (70.54%), oleic acid (4.646%), and di-N-decylsulfone (5.136%). Parallel to larvicidal assay, sub-lethal dosage acetone extracts severely affected the enzyme regulations (α,β-carboxylesterase, GST and CYP450) of third instars. Larval and pupal durations increased in all treatment sub-lethal dosage (0.127, 0.151, 0.177, and 0.197 mg/ml), whereas egg hatchability and means of fecundity decreased compared to control. The survival rate was reduced statistically in Cx. quinquefasciatus (χ² = 23.77, df = 1, P = 0.001) in all the treatment dosages as compared to the control. Antimicrobial activity assays showed significant growth inhibition post treatment with acetone and methanol extracts against Salmonella typhimurium, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus pneumoniae, Escherichia coli, and Shigella flexneri. Overall, these results indicated the potential employment of A. marina extracts as a source of natural mosquitocidal and antimicrobial compounds of green-based environment.

The gold nanoparticles (AuNPs) were synthesized using the lichen Parmelia sulcata extract (PSE) and characterized. The peaks of ultraviolet spectrophotometer and Fourier transmission infrared confirmed the formation of nanoparticles and the bioactive compounds of the lichen being responsible for reducing and capping of the particles. The face-centered cubic particles were determined by XRD peaks at 111, 200, 220, and 311. The elemental composition and spherical shape of AuNPs were confirmed by energy-dispersive spectroscopy and transmission electron microscopy. The average particle size is 54 nm, and the zeta potential − 18 was ascertained by dynamic light scattering. The potential effect of synthesized nanoparticles and lichen extracts was evaluated for antioxidant bioassays like DPPH and H₂O₂ and tested for mosquitocidal activity against Anopheles stephensi. Results showed that the lichen extract and AuNPs have the capability to scavenge the free radicals with the IC₅₀ values of DPPH being 1020 and 815 μg/ml and the IC₅₀ values of H₂O₂ being 694 and 510 μg/ml, respectively. The mosquitocidal experimental results in this study showed the inhibition of A. stephensi and A. aegypti against the larvae (I–IV instar), pupae, adult, and egg hatching. On comparison, A. stephensi showed effective inhibition than A. aegypti even at low concentration. Based on the obtained results, gold nanoparticles synthesized using PSE showed an excellent mosquitocidal effect against Anopheles stephensi.

In vitro elicitation of an important compound conessine has been done in the bark-derived callus culture of Holarrhena antidysenterica (L.) Wall. employing different elicitors. For induction of callus, green bark explants excised from field-grown plants were cultured on MS medium augmented with different concentrations (0, 1, 2.5, 5, and 10 μM) of various growth regulators such as BA, IBA, NAA, and 2,4-D either alone or in combinations. The maximum amount of conessine (458.18 ± 0.89ᵈ μg/g dry wt.) was achieved in callus developed on MS medium supplemented with 5 μM BA and 5 μM 2,4-D through HPLC analysis. Elicitation in conessine content in the above callus was achieved employing a variety of organic (phenylalanine, tyrosine, chitosan, tryptophan, casein hydrolysate, proline, sucrose, and yeast extract) as well as inorganic elicitors (Pb(NO₃)₂, As₂O₃, CuSO₄, NaCl, and CdCl₂) in different concentrations. The optimum enhancement in conessine content (3518.58 ± 0.28ᵍ μg/g dry wt.) was seen at the highest concentration (200 mg/L) of phenylalanine. The enhancement was elicitor specific and dose dependent. The overall increment of the conessine content was seen in the order of phenylalanine > tryptophan > Pb(NO₃)₂ > sucrose > NaCl > As₂O₃ > casein hydrolysate > CdCl₂ > chitosan > proline > yeast extract > CuSO₄ > tyrosine. The isolation and purification of conessine was done using methanol as a solvent system through column chromatography (CC) and TLC. The isolated compound was characterized by FT-IR, ¹H-NMR, and HRMS which confirmed with the structure of conessine. The bioassays conducted with the isolated compound revealed a strong larvicidal activity against Anopheles stephensi Liston with LC₅₀ and LC₉₀ values being 1.93 and 5.67 ppm, respectively, without harming the nontarget organism, Mesocyclops thermocyclopoides Harada, after 48 h of treatment. This is our first report for the isolation and elicitation of conessine in the callus culture of H. antidysenterica.

Recently, the biofabrication of metal nanoparticles has gained wide interest owing to its inherent features such as swift, simplicity, eco-friendliness, and cheaper costs. Different green-reducing agents led to the production of nanoparticles with varying toxicity on insects. In the current study, silver nanoparticles (AgNPs) were successfully synthesized using Habenaria plantaginea leaf extract. Ag nanoparticles were studied by UV–Vis spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), scanning electron microscopy (SEM) coupled with energy-dispersive spectroscopy (EDS), and transmission electron microscopy (TEM). H. plantaginea extract and AgNPs were tested for mosquito larvicidal activity on Anopheles stephensi, Aedes aegypti, Culex quinquefasciatus, An. subpictus, Ae. albopictus, and Cx. tritaeniorhynchus. LC₅₀ values were 102.51, 111.99, 123.47, 123.96, 136.56, 149.42 μg/ml and 12.23, 13.38, 14.78, 14.37, 15.39, 16.89 μg/ml, respectively. Moreover, H. plantaginea aqueous extract and AgNPs were tested against the non-target species Anisops bouvieri, Diplonychus indicus, Poecilia reticulata, and Gambusia affinis obtaining LC₅₀ values ranging from 831.82 to 36,212.67 μg/ml. Overall, this study showed the effectiveness of H. plantaginea-fabricated nanoparticles on a wide range of important mosquito vectors, highlighting their scarce toxicity on four natural enemies predating mosquito larvae and pupae.

Mosquitoes threaten the lives of humans, livestock, pets and wildlife around the globe, due to their ability to vector devastating diseases. Aglaia elaeagnoidea, commonly known as Priyangu, is widely employed in Asian traditional medicine and pest control. Medicinal activities include anti-inflammatory, analgesic, anticancer, and anesthetic actions. Flavaglines, six cyclopenta[b]benzofurans, a cyclopenta[bc]benzopyran, a benzo[b]oxepine, and an aromatic butyrolactone showed antifungal properties, and aglaroxin A and rocaglamide were effective to control moth pests. Here, we determined the larvicidal action of A. elaeagnoidea leaf aqueous extract. Furthermore, we focused on Priyangu-mediated synthesis of Ag nanoparticles toxic to Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. The plant extract and the nanolarvicide were tested on three mosquito vectors, following the WHO protocol, as well as on three non-target mosquito predators. Priyangu-synthesized Ag nanoparticles were characterized by spectroscopic (UV, FTIR, XRD, and EDX) and microscopic (AFM, SEM, and TEM) analyses. Priyangu extract toxicity was moderate on Cx. quinquefasciatus (LC₅₀ 246.43; LC₉₀ 462.09 μg/mL), Ae. aegypti (LC₅₀ 229.79; LC₉₀ 442.71 μg/mL), and An. stephensi (LC₅₀ 207.06; LC₉₀ 408.46 μg/mL), respectively, while Priyangu-synthesized Ag nanoparticles were highly toxic to Cx. quinquefasciatus (LC₅₀ 24.91; LC₉₀ 45.96 μg/mL), Ae. aegypti (LC₅₀ 22.80; LC₉₀ 43.23 μg/mL), and An. stephensi (LC₅₀ 20.66; LC₉₀ 39.94 μg/mL), respectively. Priyangu extract and Ag nanoparticles were found safer to non-target larvivorous fishes, backswimmers, and waterbugs, with LC₅₀ ranging from 1247 to 37,254.45 μg/mL, if compared to target pests. Overall, the current research represents a modern approach integrating traditional botanical pesticides and nanotechnology to the control of larval populations of mosquito vectors, with negligible toxicity against non-target including larvivorous fishes, backswimmers, and waterbugs.