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 recent outbreaks of dengue, chikungunya, and Zika virus highlighted the pivotal importance of mosquito vector control in tropical and subtropical areas worldwide. However, mosquito control is facing hot challenges, mainly due to the rapid development of pesticide resistance in Culicidae and the limited success of biocontrol programs on Aedes mosquitoes. In this framework, screening botanicals for their mosquitocidal potential may offer effective and eco-friendly tools in the fight against mosquitoes. In the present study, the essential oil (EO) obtained from the medicinal plant Origanum scabrum was analyzed by GC-MS and evaluated for its mosquitocidal and repellent activities towards Anopheles stephensi, Aedes aegypti, Culex quinquefasciatus, and Culex tritaeniorhynchus. GC-MS analysis showed a total of 28 compounds, representing 97.1 % of the EO. The major constituents were carvacrol (48.2 %) and thymol (16.6 %). The EO was toxic effect to the A. stephensi, A. aegypti, C. quinquefasciatus, and C. tritaeniorhynchus larvae, with LC₅₀ of 61.65, 67.13, 72.45, and 78.87 μg/ml, respectively. Complete ovicidal activity was observed at 160, 200, 240, and 280 μg/ml, respectively. Against adult mosquitoes, LD₅₀ were 122.38, 134.39, 144.53, and 158.87 μg/ml, respectively. In repellency assays, the EOs tested at 1.0, 2.5, and 5.0 mg/cm² concentration of O. scabrum gave 100 % protection from mosquito bites up to 210, 180, 150, and 120 min, respectively. From an eco-toxicological point of view, the EO was tested on three non-target mosquito predators, Gambusia affinis, Diplonychus indicus, and Anisops bouvieri, with LC₅₀ ranging from 4162 to 12,425 μg/ml. Overall, the EO from O. scabrum may be considered as a low-cost and eco-friendly source of phytochemicals to develop novel repellents against Culicidae.

Silver nanoparticles (AgNPs) were successfully synthesised from aqueous silver nitrate using the extracts of Arachis hypogaea peels. The synthesised SNPs were characterized by Fourier transform-infrared spectroscopy analysis, X-ray diffraction, transmission electron microscopy analysis and high-resonance scanning electron microscopy, and energy dispersive X-ray spectroscopy. AgNPs were well defined and measured 20 to 50 nm in size. The nanoparticles were crystallized with a face-centered cubic structure. Larvicidal activity of synthesised AgNPs from A. hypogaea peels was tested for their larvicidal activity against the fourth instar larvae of Aedes aegypti (Yellow fever), Anopheles stephensi (Human malaria). The results suggest that the synthesised AgNPs have the potential to be used as an ideal eco-friendly resource for the control of A. aegypti and A. stephensi. This study provides the first report on the mosquito larvicidal activity of synthesised AgNPs from A. hypogaea peels against vectors of malaria and dengue.

In mosquito control programs, insecticides of botanical origin have the potential to eliminate eggs, larvae, and adults. So, the larvicidal, ovicidal, and oviposition-deterrent activities of petroleum ether and ethyl acetate extracts of the leaves of Eugenia jambolana, Solidago canadensis, Euodia ridleyi, and Spilanthes mauritiana were assayed against the three vector mosquito species, namely Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The larval bioassay was conducted following the World Health Organization method. The maximum larval mortality was found with ethyl acetate extract of S. mauritiana against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus with LC₅₀values of 11.51, 28.1, 14.10 ppm, respectively. The mean percent hatchability of the ovicidal activity was observed at 48-h post-treatment. The percent hatchability was found to be inversely proportional to the concentration of the extract and directly proportional to the number of eggs. The flower head extract of S. mauritiana gave 100 % mortality followed by E. ridleyi, S. canadensis, and E. jambolana against the eggs of the three mosquito vectors. For oviposition-deterrent effect, out of the five concentrations tested (20, 40, 60, 80, and 100 ppm), the concentration of 100 ppm showed a significant egg laying-deterrent capacity. The oviposition activity index value of E. jambolana, E. ridleyi, S. canadensis, and S. mauritiana against A. aegypti, A. stephensi, C. quinquefasciatus at 100 ppm were −0.71, −0.71, −0.90, −0.93, −0.85, −0.91, −1, −1, −0.71, −0.85, −1, and −1, respectively. These results suggest that the leaf/flower extracts of certain local plants have the potential to be developed as possible eco-friendly means for the control of mosquitoes.

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.

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.

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.

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.

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.

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.