Knowledge about how the COVID-19 pandemic can affect aquatic wildlife is still extremely limited, and no effect of SARS-CoV-2 or its structural constituents on invertebrate models has been reported so far. Thus, we investigated the presence of the 2019-new coronavirus in different urban wastewater samples and, later, evaluated the behavioral and biochemical effects of the exposure of Culex quinquefasciatus larvae to two SARS-CoV-2 spike protein peptides (PSPD-2002 and PSPD-2003) synthesized in our laboratory. Initially, our results show the contamination of wastewater by the new coronavirus, via RT-qPCR on the viral N1 gene. On the other hand, our study shows that short-term exposure (48 h) to a low concentration (40 μg/L) of the synthesized peptides induced changes in the locomotor and the olfactory-driven behavior of the C. quinquefascitus larvae, which were associated with increased production of ROS and AChE activity (cholinesterase effect). To our knowledge, this is the first study that reports the indirect effects of the COVID-19 pandemic on the larval phase of a freshwater invertebrate species. The results raise concerns at the ecological level where the observed biological effects may lead to drastic consequences.

The increasing demand for fresh water has forced many countries to use reclaimed wastewater for agricultural purposes. This water contains pharmaceuticals and personal care products (PPCPs) that remain biologically active following passage through wastewater treatment plants. Run-off from farms and contaminated water from treatment facilities exposes aquatic ecosystems to PPCPs. This study examined the effects of PPCPs on a lower trophic organism. Culex quinquefasciatus larvae were reared in water contaminated with environmentally relevant concentrations of common PPCPs. Acetaminophen alone and a mixture of contaminants were found to increase developmental time of larvae. Susceptibility to Bti increased in larvae exposed to antibiotics, acetaminophen, or a mixture of PPCPs. Antibiotics, hormones, and the mixture altered the mosquito bacterial microbiome. Overall, the results indicate that at environmentally relevant concentrations, PPCPs in reclaimed water can have biologically important effects on an ecologically and medically important lower trophic level insect.

The essential oil (EO) of Cupressus sempervirens was obtained by hydrodistillation and analysed using gas chromatography–flame ionization detection (GC–FID) and gas chromatography–mass spectrometry (GC–MS). Two monoterpenes, α-pinene (49.1%) and δ-3-carene (21.4%), and one sesquiterpene hydrocarbon, α-cedrol (5.1%), were isolated as the EO major terpenes. An oil-in-water nanoemulsion (particle size 71.2 nm) was produced from the EO through a low-energy method. The EO, its nanoemulsion and its main constituents showed mosquitocidal and biochemical effects against Culex quinquefasciatus Say, the common vector of lymphatic filariasis parasites. All treatments showed dose-dependent bioactivity, and adults were more susceptible to the EO products than the larvae. The nanoemulsion showed superior activity, followed by the crude EO and α-cedrol. At 40 μg/ml, the nanoemulsion caused 100% larval mortality, while the EO and α-cedrol required twice this concentration to achieve the same larval mortality. The LC₅₀ values were 8.4, 16.1, 15.1, 30.7 and 53.4 μg/ml at 24 h after exposure for the nanoemulsion, crude oil, α-cedrol, δ-3-carene and α-pinene, respectively. For adults, 20.0 μl/l nanoemulsion caused 100% mortality, while twice this concentration of the EO was required to achieve the same effect. The LC₅₀’s against adults ranged between 6.2 and 40.4 μl/l. EO products prominently repelled mosquitoes at concentrations between 0.75 and 6.0 μl/cm². The EO products caused remarkable inhibition of Cx. quinquefasciatus acetylcholinesterase activity but were safer towards the non-target aquatic species Gambusia affinis. These results recommend the use of C. sempervirens EO, its nanoemulsion and main terpenes as natural tools to control Cx. quinquefasciatus.

Slaughterhouse waste water (SHW) is discarded as unused disposals into the environment every day. The objective of the present study is to explore the possibility on the utilization of SHW for the preparation of culture media to produce mosquitocidal bacterium (Bacillus thuringiensis subsp. israelensis). This would help to overcome the problem of dumping SHW in to environment. A judicial combination of SHW with a mineral salt (MnCl2) was made to produce an enhanced level of bacterial production when compared with other culture media including conventional medium (Luria Bertani, LB). A complete degradation of SHW by the bacteria was observed. The biomass yield, bacterial growth, toxin production, and larvicidal activity against mosquito vectors were satisfactory. Cell mass yield of 4.55 gm l−1 (dry wt) and larvicidal activity of 0.006 mg ml−1 and 0.026 mg ml−1 at LC50 and LC90 levels were observed, respectively, against the filarial vector of Culex quinquefasciatus with bacteria grown in SHW + MnCl2. The B. thuringiensis subsp. israelensis also controlled the larvae in the field significantly for three weeks (>90% mortality) and the effect was comparable with LB. Cost-analysis for production of B. thuringiensis subsp. israelensis showed that it is more economical. Thus, this study suggested the dual benefit of efficient production of mosquitocidal toxin and management of slaughterhouse wastewater.

Medicinal and aromatic plants represent an outstanding source of green active ingredients for a broad range of real-world applications. In the present study, we investigated the insecticidal potential of the essential oils obtained from three medicinal and aromatic plants of economic importance in Algeria, Artemisia campestris, Pulicaria arabica, and Saccocalyx satureioides. Gas chromatography coupled with mass spectrometry (GC-MS) was used to study the essential oil chemical compositions. The three essential oils were tested against a mosquito vectoring filariasis and arboviruses, i.e., Culex quinquefasciatus, a fly pest acting also as pathogens vector, Musca domestica, and an agricultural moth pest, i.e., Spodoptera littoralis, using WHO and topical application methods, respectively. The essential oil from A. campestris, containing β-pinene (15.2%), α-pinene (11.2%), myrcene (10.3%), germacrene D (9.0%) (Z)-β-ocimene (8.1%) and γ-curcumene (6.4%), showed remarkable toxicity against C. quinquefasciatus (LC₅₀ of 45.8 mg L⁻¹) and moderate effects (LD₅₀ of 99.8 μg adult⁻¹) against M. domestica. Those from P. arabica and S. satureioides, containing epi-α-cadinol (23.9%), δ-cadinene (21.1%), α-cadinol (19.8%) and germacrene D-4-ol (8.4%), and thymol (25.6%), α-terpineol (24.6%), borneol (17.4%) and p-cymene (11.4%), respectively, were more active on S. littoralis showing LD₅₀ values of 68.9 and 61.2 μg larva⁻¹, respectively. Based on our results, the essential oil from A. campestris may be further considered a candidate ingredient for developing botanical larvicides.

Populations of Culex quinquefasciatus Say, 1823 (Diptera: Culicidae) have shown resistance to insecticides of the carbamate and organophosphate classes. The objective of this study was to assess the susceptibility of C. quinquefasciatus larvae to essential oils from leaves of Eugenia uniflora L., Melaleuca armillaris (Sol. ex Gaertn.) Sm., and Schinus molle L and C. quinquefasciatus larvae’s biochemical responses after their exposure to these leaves. The essential oils were chemically analyzed by GC and GC/MS. First, the lethal concentration for 50% (LC₅₀) values was estimated using different concentrations of essential oils and probit analysis. The larvae were exposed for 1 h at the LC₅₀ estimated for each essential oil. The susceptibility of the larvae to essential oils was evaluated using the following biochemical parameters: concentrations of total protein and reduced glutathione; levels of production of hydrogen peroxide and lipid peroxidation; and the activity of the enzyme acetylcholinesterase (AChE). The main chemical constituents in E. uniflora were E-β-ocimene, curzerene, germacrene B, and germacrone; in M. armillaris were 1,8-cineole and terpinolene; and in S. molle were sabinene, myrcene, and sylvestrene. The essential oils had LC₅₀ values between 31.52 and 60.08 mg/L, all of which were considered effective. All of them also promoted changes in biochemical parameters when compared to the control treatment. The essential oils of S. molle and E. uniflora inhibited the activity of the AChE enzyme, and the essential oil of M. armillaris increased it. All essential oils had larvicidal activity against C. quinquefasciatus, but the essential oil of E. uniflora was the most efficient. Thus, the findings of the present study suggest that the essential oil of E. uniflora can be considered promising for the development of botanical larvicides.

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.

Titanium dioxide nanoparticles are emerging as a biocompatible nanomaterial with multipurpose bioactivities. In this study, titanium dioxide (TiO₂) nanoparticles were effectively synthesized using the aqueous leaf extracts of Parthenium hysterophorus prepared by microwave irradiation. TiO₂ nanoparticles were fabricated by treating the P. hysterophorus leaf extracts with the TiO₄ solution. Biologically active compounds such as alcohols, phenols, alkanes, and fluoroalkanes were involved in bioreduction of TiO₄ into TiO₂. The formation of green-engineered TiO₂ nanoparticles was confirmed by UV-vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), field emission scanning electron microscopy (FESEM), and energy-dispersive X-ray (EDX) spectroscopy and further characterized by X-ray diffraction (XRD) studies. UV-vis spectroscopy analysis showed maximum absorbance at 420 nm due to surface plasmon resonance of synthesized TiO₂ NPs. FTIR spectrum of the engineered TiO₂ NPs showed the presence of bioactive compounds in the leaf extract, which acted as capping and reducing agents. FESEM exhibited an average size of 20–50 nm and a spherical shape of TiO₂ NPs. EDX analysis indicated the presence of TiO₂ NPs by observing the peaks of titanium ions. XRD results pointed out the crystalline nature of engineered TiO₂ NPs. The larvicidal activity of TiO₂ NPs was studied on fourth instar larvae of dengue, Zika virus, and filariasis mosquito vectors Aedes aegypti and Culex quinquefasciatus. Antimicrobial efficacy of TiO₂ NPs was assessed on clinically isolated pathogens Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus vulgaris, and Staphylococcus epidermidis. Besides, we found that TiO₂ NPs are able to quickly degrade the industrially harmful pigments methylene blue, methyl orange, crystal violet, and alizarin red dyes under sunlight illumination. Overall, this novel, simple, and eco-friendly approach can be of interest for the control of vector-borne diseases, as well as to formulate new bactericidal agents and to efficiently degrade dye solutions in the polluted areas.

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.