North-Eastern Brazil saw intensive application of the insecticide pyriproxyfen (PPF) during the microcephaly outbreak caused by the Zika virus (ZIKV). ZIKV requires the neural RNA-binding protein Musashi-1 to replicate. Thyroid hormone (TH) represses MSI1. PPF is a suspected TH disruptor. We hypothesized that co-exposure to the main metabolite of PPF, 4′–OH–PPF, could exacerbate ZIKV effects through increased MSI1 expression. Exposing an in vivo reporter model, Xenopus laevis, to 4′–OH–PPF decreased TH signaling and increased msi1 mRNA and protein, confirming TH-antagonistic properties. Next, we investigated the metabolite's effects on mouse subventricular zone-derived neural stem cells (NSCs). Exposure to 4′–OH–PPF dose-dependently reduced neuroprogenitor proliferation and dysregulated genes implicated in neurogliogenesis. The highest dose induced Msi1 mRNA and protein, increasing cell apoptosis and the ratio of neurons to glial cells. Given these effects of the metabolite alone, we considered if combined infection with ZIKV worsened neurogenic events. Only at the fourth and last day of incubation did co-exposure of 4′–OH–PPF and ZIKV decrease viral replication, but viral RNA copies stayed within the same order of magnitude. Intracellular RNA content of NSCs was decreased in the combined presence of 4′–OH–PPF and ZIKV, suggesting a synergistic block of transcriptional machinery. Seven out of 12 tested key genes in TH signaling and neuroglial commitment were dysregulated by co-exposure, of which four were unaltered when exposed to 4′–OH–PPF alone. We conclude that 4′–OH–PPF is an active TH-antagonist, altering NSC processes known to underlie correct cortical development. A combination of the TH-disrupting metabolite and ZIKV could aggravate the microcephaly phenotype.

This paper explores the main factors for mosquito-borne transmission of the Zika virus by focusing on environmental, anthropogenic, and social risks. A literature review was conducted bringing together related information from this genre of research from peer-reviewed publications. It was observed that environmental conditions, especially precipitation, humidity, and temperature, played a role in the transmission. Furthermore, anthropogenic factors including sanitation, urbanization, and environmental pollution promote the transmission by affecting the mosquito density. In addition, socioeconomic factors such as poverty as well as social inequality and low-quality housing have also an impact since these are social factors that limit access to certain facilities or infrastructure which, in turn, promote transmission when absent (e.g., piped water and screened windows). Finally, the paper presents short-, mid-, and long-term preventative solutions together with future perspectives. This is the first review exploring the effects of anthropogenic aspects on Zika transmission with a special emphasis in Brazil.

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.

Dengue is the most rapidly spreading mosquito-borne viral disease in the world. The mosquito, Aedes aegypti, also spreads Yellow fever, Chikungunya, and Zika virus. As the primary vector for dengue, Ae. aegypti now occurs in over 20 countries and is a serious concern with reports of increasing insecticide resistance. Developing new treatments to manage mosquitoes are needed. Formulation of crude volatile oil from Piper betle leaves (Pb-CVO) was evaluated as a potential treatment which showed larvicidal, ovipositional, and repellency effects. Gut-histology and enzyme profiles were analyzed post treatment under in-vitro conditions. The Pb-CVO from leaves of field collected plants was obtained by steam distillation and separated through rotary evaporation. The Pb-CVO were evaluated for chemical constituents through GC-MS analyses revealed 20 vital compounds. The peak area was establish to be superior in Eudesm-7(11)-en-4-ol (14.95%). Pb-CVO were determined and tested as four different concentrations (0.25, 0.5, 1.0, and 1.5 mg/L) of Pb-CVO towards Ae. aegypti. The larvicidal effects exhibited dose dependent mortality being greatest at 1.5 mg Pb-CVO/10 g leaves. The LC₅₀ occurred at 0.63 mg Pb-CVO/L. Larva of Ae. aegypti exposed to Pb-CVO showed significantly reduced digestive enzyme actions of α- and β-carboxylesterases. In contrast, GST and CYP450 enzyme levels increased significantly as concentration increased. Correspondingly, oviposition deterrence index and egg hatch of Ae. aegypti exposed to sub-lethal doses of Pb-CVO demonstrated a strong effect suitable for population suppression. Repellency at 0.6 mg Pb-CVO applied as oil had a protection time of 15–210 min. Mid-gut histological of Ae. aegypti larvae showed severe damage when treated with 0.6 mg of Pb-CVO treatment compared to the control. Non-toxic effects against aquatic beneficial insects, such as Anisops bouvieri and Toxorhynchites splendens, were observed at the highest concentrations, exposed for 3 h. These results suggest that the Pb-CVO may contain effective constituents suitable for development of new vector control agents against Ae. aegypti.

Microbial polysaccharides produced by marine species play a key role in food and cosmetic industry, as they are nontoxic and biodegradable polymers. This investigation reports the isolation of exopolysaccharide from Bacillus licheniformis Dahb1 and its biomedical applications. Bacillus licheniformis Dahb1 exopolysaccharide (Bl-EPS) was extracted using the ethanol precipitation method and structurally characterized. FTIR and ¹H-NMR pointed out the presence of various functional groups and primary aromatic compounds, respectively. Bl-EPS exhibited strong antioxidant potential confirmed via DPPH radical, reducing power and superoxide anion scavenging assays. Microscopic analysis revealed that the antibiofilm activity of Bl-EPS (75 μg/ml) was higher against Gram-negative (Pseudomonas aeruginosa and Proteus vulgaris) bacteria over Gram-positive species (Bacillus subtilis and Bacillus pumilus). Bl-EPS led to biofilm inhibition against Candida albicans when tested at 75 μg/ml. The hemolytic assay showed low cytotoxicity of Bl-EPS at 5 mg/ml. Besides, Bl-EPS achieved LC₅₀ values < 80 μg/ml against larvae of mosquito vectors Anopheles stephensi and Aedes aegypti. Overall, our findings pointed out the multipurpose bioactivity of Bl-EPS, which deserves further consideration for pharmaceutical, environmental and entomological applications.

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.

Pyriproxyfen is used in Brazil to combat epidemics of Dengue Fever, Chikungunya Fever, and Zika virus. This study assessed the effects of pyriproxyfen on reproductive performance, embryo-fetal development, head measurements, and DNA integrity in a preclinical model. Thirty pregnant mice were divided into three groups (n = 10): control (drinking water—0.1 ml/10 g (body weight—b.w., gavage) and treated with pyriproxyfen 0.0002 mg/kg and 0.0021 mg/kg (b.w., gavage) during the gestational period. Analysis of biometric, reproductive performance and embryo-fetal development parameters related to control presented no significant differences, suggesting no maternal or embryo-fetal toxicity. Head measurements showed no differences except an increase in anterior/posterior measurement and glabella/external occipital protuberance. Analysis of DNA integrity showed an increase in micronucleus only at 72 h for the lowest dose group. Thus, we infer that pyriproxyfen is not related to the occurrence of microcephaly, nor does it alter reproductive performance, embryo-fetal development or DNA integrity.

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.

Plant extracts with their enriched chemical constituents have established potential alternative mosquito control agents. In this research, we developed an eco-friendly mosquitocidal agent from Alangium salvifolium leaves against the dengue and Zika virus vector Aedes aegypti and we investigated its biosafety on the mosquito aquatic predator Toxorhynchites splendens. Results showed that the methanolic extract of A. salvifolium leaves was composed by eight main compounds, with major peak area for hexadecenoic acid (21.74%). LC₅₀ and LC₉₀ values calculated on Ae. aegypti fourth instar larvae were 104.80 and 269.15 ppm respectively. The methanolic extract tested at 100 ppm decreased the α-β carboxylesterase and SOD ratio significantly and upregulated the GST and CYP450 level. The A. salvifolium methanolic extract displayed significant repellent and adulticidal activity at 100 and 400 ppm respectively. The treatment with 100 ppm of the methanolic extract led to 210 min of protection from Ae. aegypti bites. Four hundred parts per million of the extract showed 98% adult mortality within 30 min from the treatment. Lastly, biosafety assays on the mosquito aquatic predator Tx. splendens showed that the toxicity of the A. salvifolium extract was significantly lower if compared to the cypermethrin-based treatments. The methanolic extract of A. salvifolium showed a maximum of 47.3% mortality rate at the concentration of 1000 ppm, while 0.7 ppm of cypermethrin achieved 91.3% mortality rate on Tx. splendens. Overall, our study enhances basic knowledge on how to improve natural larvicidal agents against dengue and Zika virus mosquito vector with harmless responses on non-target aquatic predators.

Aedes aegypti is the vector responsible for transmitting pathogens that cause various infectious diseases, such as dengue, Zika, yellow fever, and chikungunya, worrying health authorities in the tropics. Due to resistance of mosquitoes to synthetic insecticides, the search for more effective insecticidal agents becomes crucial. The aim of this study was to verify the larvicidal, adulticidal, and anticholinesterase activities of the essential oils of the Illicium verum (EOIV), Pimenta dioica (EOPD), and Myristica fragrans (EOMF) against Ae. aegypti. The essential oils (EOs) were obtained by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS). The larvicidal and adulticidal activities of EOs were evaluated against third instar larvae and Ae. aegypti adult females, respectively, using the procedures of the World Health Organization (WHO) and the anticholinesterase activity of the EOs by the modified Ellman method. The following major components were identified: (E)-anethole (90.1%) for EOIV, methyl eugenol (55.0%) for EOPD, and sabinene (52.1%) for EOMF. All EOs exhibited larvicidal and adulticidal activity against Ae. aegypti. The highest larval mortality was observed in EOMF with LC₅₀ = 28.2 μg mL⁻¹. Adult mortality was observed after 1 (knockdown) and 24 h exposure, with the highest potential established by the EOIV, KC₅₀ = 7.3 μg mg female⁻¹ and LC₅₀ = 10.3 μg mg female⁻¹. EOIV (IC₅₀ = 4800 μg mL⁻¹), EOMF (IC₅₀ = 4510 μg mL⁻¹), and EOPD (IC₅₀ = 1320 μg mL⁻¹) inhibited AChE. EOMF (4130 μg mL⁻¹) and EOPD (IC₅₀ = 3340 μg mL⁻¹) inhibited BChE whereas EOIV showed no inhibition. The EOs were toxic to larvae and adults of Ae. aegypti, as well as being less toxic to humans than the currently used insecticides, opening the possibility of elaboration of a natural, safe, and ecological bioinsecticide for vector control.