Larvicidal effects of some essential oils against Aedes aegypti (L.), the vector of dengue fever in Saudi Arabia

Essential oils are very popular among organic growers because they are ecologically safe, do not have mammalian toxicity, and cannot be resistant to a variety of contaminants. Four essential oils, Lemon, Lavender, Peppermint, and Neem, were tested for larvicide efficacy against the dengue fever vector Aedes aegypti larvae under laboratory conditions using dipping bioassay techniques. Among the essential oils tested, lemon, peppermint, and lavender oils showed high larvicidal activity against larvae of Ae. aegypti. Lemon oil showed the highest effects (LC₅₀ 10.676 ppm), while Peppermint, Lavender and Neem oil showed the lowest effects (LC₅₀ 21.380, 29.818 and 38.058 ppm, respectively). As a result, the mixture of lemon oil (LC₅₀) with Peppermint oil (LC₂₅) showed the highest co-toxicity factor, whereas the mixture of Lemon oil (LC₅₀) with Diesel oil (LC₂₅) showed the lowest co-toxicity factor. Based on the results of this study, it appears that essential oils may be useful as larvicides against Ae. aegypti larvae. In search of new natural larvicides, these compounds may provide an alternative to Synthetic insecticides as these are environmentally safe insecticides.     

Larvicidal and adulticidal activity of essential oils from plants of the Lamiaceae family against the West Nile virus vector,Culex pipiens (Diptera: Culicidae)

Culex pipiens mosquitoes are the most widely distributed primary vector of the West Nile virus worldwide. Many attempts for investigation of botanical pesticides to avoid the development of pesticide resistance to conventional synthetic pesticides that are recognized as a threat to the diversity of ecosystems. The study aimed to determine the components of three essential oils of Lamiaceae family, lavender (Lavandula angustifolia), peppermint (Mentha piperita L.), and rosemary (Rosmarinus officinalis L.) by gas chromatography-mass spectrometry (GC–MS) analysis. Furthermore, aimed to validate the insecticidal activities of these oils as larvicidal agents against the third instar larvae of Culex pipiens using five different concentrations (62.5, 125, 250, 500, and 1000 ppm) for each oil in five replicates and as an adulticidal agent against approximately three-day-old female adults of Cx. Pipiens using 0.5, 1, 2, 4, and 5% concentrations in three replicates. The results generally showed a dose-related response. At 1000 ppm, rosemary oil showed the highest larvicidal (100%) (LC₅₀, 214.97 ppm), followed by peppermint oil (92.00% mortality and LC₅₀ (269.35 ppm). Lavender oil showed the lowest efficacy with 87.20% mortality and LC₅₀ (301.11 ppm). At 5% oil concentration, the highest knockdown rate at 1 h was recorded for lavender oil (95.55%), followed by peppermint oil (88.89%) and lastly rosemary oil (84.44%). After 24 h, rosemary oil showed the lowest adult mortality rate (88.89%; LC₅₀, 1.44%), while lavender and peppermint oils both showed a 100% mortality rate, with (LC₅₀, 0.81% and 0.91%, respectively). The chemical constituents of the oils consisted of monoterpenes and sesquiterpenes that determined their insecticidal activities against the target insect stage. The study proposed that rosemary essential oil may be useful for the control of Cx. pipiens larvae as part of an integrated water treatment strategy, and lavender and peppermint oils may be used in an integrated plan for adult’s control.     

Comparative study of three herbal formulations against dengue vectors Aedes aegypti

The efficacy of three formulations (i.e., natural lavender crude, essential oil, and gel) extracted from Lavender angustifolia was tested against vectors of the epidemic dengue virus, Aedesaegypti, to evaluate their larvicidal activity effect. The ethanolic extract of the lavender crude was prepared using a rotary evaporator, while the other extracts, such as essential oil and gel, were obtained from iHerb, a supplier of medicinal herbs in the US. The mortality rate of larvae was evaluated 24 h after exposure. Larvicidal activity of the lavender crude was 91% mortality at 150 ppm, 94% for essential oil at a concentration of 3000 ppm, and 97% for lavender gel at a 1000 ppm. Natural lavender crude was one of the most promising extracts tested against Ae.aegypti larvae, with lethal concentrations at LC₅₀ and LC₉₀ of 76.4 and 174.5 ppm post-treatment. The essential oil had the least effect on mosquito larvae, with LC₅₀ and LC₉₀ reaching 1814.8 and 3381.9 ppm, respectively. The lavender gel was moderately effective against Ae. aegypti larvae, with LC₅₀ and LC₉₀ values reaching 416.3 and 987.7 ppm after exposure. The occurrence of morphological abnormalities in the larvae treated with the three compounds, in turn, resulted in an incomplete life cycle. Therefore, our results indicated that natural lavender crude displayed the highest larvicidal activity against larvae, followed by gel and essential oil. Thus, this study concluded that lavender crude is an effective, eco-friendly compound that can be used as an alternative to chemical products to control vector-borne epidemic diseases.     

Larvicidal activity of some fungal extracts on Aedes caspius and Culex pipiens (Diptera: Culicidae)

A preliminary study was conducted to evaluate the effects of nine methanol fungal extracts on fourth instar larvae of Aedes caspius and Culex pipiens (Diptera: Culicidae). None of the extracts tested showed larvicidal activity except for the Paecilomyces lilacinus extract, which showed 100 % mortality against the larvae of Ae caspius after 24 h. The LC₅₀ values of the P. lilacinus extract after 24 h were 190.66 μg/mL against Ae. caspius and 254.25 μg/mL against Cx. pipiens, respectively. After 48 h, meanwhile, the LC₅₀ values were 65.70 and 164.13 μg/mL, respectively. Histological analysis of the midgut of the treated larvae of Ae. caspius and Cx. pipiens revealed changes such as cell destruction, spacing between the cells and disruption of the microvilli, resulting in an appearance of vacuolization in the midgut. This article is the first report of the use of P. lilacinus extract for the control of Ae. caspius and Cx. pipiens larvae and the data obtained may help to provide a better understanding of the mode of action of P. lilacinus as an insecticide against mosquito larvae.     

Mosquito larvicidal activity of Aloe vera (Family: Liliaceae) leaf extract and Bacillus sphaericus,against Chikungunya vector,Aedes aegypti

The bio-efficacy of Aloe vera leaf extract and bacterial insecticide, Bacillus sphaericus larvicidal activity was assessed against the first to fourth instars larvae of Aedes aegypti, under the laboratory conditions. The plant material was shade dried at room temperature and powdered coarsely. A. vera and B. sphaericus show varied degrees of larvicidal activity against various instars larvae of A. aegypti. The LC₅₀ of A. vera against the first to fourth instars larvae were 162.74, 201.43, 253.30 and 300.05 ppm and the LC₉₀ 442.98, 518.86, 563.18 and 612.96 ppm, respectively. B. sphaericus against the first to fourth instars larvae the LC₅₀ values were 68.21, 79.13, 93.48, and 107.05 ppm and the LC₉₀ values 149.15, 164.67, 183.84, and 201.09 ppm, respectively. However, the combined treatment of A. vera + B. sphaericus (1:2) material shows highest larvicidal activity of the LC₅₀ values 54.80, 63.11, 74.66 and 95.10 ppm; The LC₉₀ values of 145.29, 160.14, 179.74 and 209.98 ppm, against A. aegypti in all the tested concentrations than the individuals and clearly established that there is a substantial amount of synergist act. The present investigation clearly exhibits that both A. vera and B. sphaericus materials could serve as a potential larvicidal agent. Since, A. aegypti is a container breeder vector mosquito this user and eco-friendly and low-cost vector control strategy could be a viable solution to the existing dengue disease burden. Therefore, this study provides first report on the mosquito larvicidal activity the combined effect of A. vera leaf extract and B. sphaericus against as target species of A. aegypti.     

Pathogenicity of entomopathogenic fungus,Metarhizium anisopliae MET-GRA4 isolate on dengue vectors,Aedes albopictus and Aedes aegypti mosquito larvae (Diptera: Culicidae)

Considering the rapid transmission of the dengue virus, substantial efforts need to be conducted to ward-off the epidemics of dengue viruses. The control effort is depending on chemical insecticides and had aroused undesirable conflicts of insecticide resistance. Here, we study the entomopathogenic fungus, Metarhizium anisopliae as a promising new biological control agent for vector control. The pathogenicity effects of Metarhizium anisopliae against field and laboratory strains of Aedes albopictus and Aedes aegypti larvae were tested using the larvicidal bioassay technique. The results demonstrate that the treatments using M. anisopliae isolate MET-GRA4 were highly effective and able to kill 100% of both Ae. albopictus and Ae. aegypti mosquito larvae at a conidia concentration of 1 × 10?/ml within 7 days of the treatment period. The fungus displayed high larvicidal activity against laboratory and field strain of Ae. aegypti larvae with LC₅₀ values (9.6 × 10³/ml, 1.3 × 10³/ml) and LC₉₅ values (1.2 × 10?/ml, 5.5 × 10⁵/ml) respectively. For Ae. albopictus, LC₅₀ values for laboratory and field strains were (1.7 × 10⁴/ml, 2.7 × 10⁴/ml) and the LC₉₅ values were (2.1 × 10?/ml, 7.0 × 10⁵/ml) respectively. Interestingly, the susceptibility of field strain towards M. anisopliae was higher as compared to the laboratory strain Aedes larvae. In which, the causative agents of all the dead larvae were verified by the virulence of M. anisopliae and caused morphological deformities on larval body. The findings from this study identify this isolate could be an effective potential biocontrol agent for vector mosquitoes in Malaysia.     

Chemical composition,oviposition deterrent and larvicidal activities against Aedes aegypti of essential oils from Piper marginatum Jacq. (Piperaceae)

The essential oils of leaves, stems and inflorescences of Piper marginatum, harvested in the Atlantic forest in the State of Pernambuco, Brazil, were obtained by hydrodistillation. GC and GC–MS analyses revealed the presence of 40 components accounting, respectively, for 99.6%, 99.7% and 99.1% of the leaf, stem and inflorescence oil, the most abundant being (Z)- or (E)-asarone and patchouli alcohol. The essential oil of the inflorescences exhibited potent activity against the 4th instar of Aedes aegypti with LC₁₀ and LC₅₀ values of 13.8 and 20.0 ppm, respectively. Furthermore, the inflorescence oil did not interfere in the oviposition of A. aegypti females when assayed at 50 ppm. These properties suggest that P. marginatum oil is a potential source of valuable larvicidal compounds for direct use or in conjunction with baits in traps constructed to capture eggs and larvae.     

Biological activity of Xanthium strumarium seed extracts on different cancer cell lines and Aedes caspius,Culex pipiens (Diptera: Culicidae)

Effects of methanol extracts of Xanthium strumarium on different cancer cell lines and on the mortality rates of Aedes caspius, Culex pipiens (Diptera: Culicidae) were investigated. Among the cell lines tested, the Jurkat cell line was the most sensitive to the methanol extract and ethyl acetate fraction, with reported LC₅₀ values of 50.18 and 48.73 μg/ml respectively. Conversely, methanol extracts were not that toxic to the A549 cell line though the toxicity increased on further purification. The percentage of growth inhibition was dose dependent for the methanol extract and ethyl acetate fraction. The ethyl acetate fraction showed higher toxicity to all cell lines tested when compared to the methanol extract. The results showed that methanol extracts of plant seeds caused 100% mortality of mosquito larvae at a concentration of 1000 μg/ml after 24 h of treatment. The LC₅₀ and LC₉₀ values of X. strumarium were found to be 531.07 and 905.95 μg/ml against Ae. caspius and 502.32 and 867.63 μg/ml against Cx. Pipiens, respectively. From the investigations, it was concluded that the crude extract of X. strumarium showed a weak potential for controlling the larval instars of Ae. caspius and Cx. pipiens. However, on further purification the extract lost the larvicidal activity. The ethyl acetate fraction showed higher toxicity to all cell lines tested when compared to the methanol extract. The ethyl acetate fraction investigated in this study appears to have a weak larvicidal activity but a promising cytotoxic activity. Future studies will include purification and investigation in further detail of the action of X. strumarium on Cancer Cell Lines and mosquitoes.     

Evaluation of larvicidal efficacy of Ricinus communis (Castor) and synthesized green silver nanoparticles against Aedes aegypti L.

Aedes mosquitoes are the most important group of vectors that transmit pathogens, including arboviruses, and cause human diseases such as dengue fever, yellow fever, Zika virus, and Chikungunya. Biosynthesis and the use of green silver nanoparticles (AgNPs) is a vital step to identify reliable and eco-friendly controls for these vectors. In this study, Aedes (Ae.) aegypti larvae (2nd and 3rd instar) were exposed to leaf extracts of Ricinus communis (Castor) and AgNPs synthesized from the extract to evaluate their larvicidal potential. Synthesized AgNPs were characterized by UV–Vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), and energy-dispersive X-ray spectroscopy (XRD). Ae. aegypti larvae were treated with different concentrations (50–250 ppm) of the leaf extract and synthesized AgNPs. There were five replicates per treatment, in addition to a positive (temephos) and negative control (dechlorinated water). Mortality was recorded after 12, 24, 36, and 48 h and the data were subjected to Probit analysis. The nanoparticles were more toxic (LC₅₀ = 46.22 ppm and LC₉₀ = 85.30 ppm) than the plant extract (106.24 and 175.73 ppm, respectively). The leaf extracts of Ricinus communis were subjected to HPLC analysis to identify their chemical constituents. This study suggests that plant extracts and synthesized nanoparticles are excellent alternatives to hazardous chemical pesticides used to control vector mosquitoes. This is a potentially useful technique that can reduce aquatic toxicity from insecticide use.     

Biosynthesis of gold and silver nanoparticles using Senna alexandrina Miller extract against Culex pipiens L. (Diptera: Culicidae)

The use of nanoparticles for various purposes, including pest control, has become increasingly popular because of their cost and environmental safety. In the present study, gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) were synthesized in an extract of Senna alexandrina Miller leaves with the aim of use against vectors of disease such as Culex pipiens L. (the filarial vector in Saudi Arabia). The nanoparticles were characterized by scanning electron microscopy and spectroscopic techniques. The larvicidal activity of the nanoparticles against Cx. pipiens was evaluated according to the protocol of the World Health Organization. According to the lethal concentration LC₅₀, the result shows differentiation in the sensitivity on mosquitoes. The AuNPs (51.383 ppm) the best one followed by AgNPs (52.525 ppm) while S. alexandrina leaf extract alone (355.25 ppm), the lowest effectiveness. Generally, the Cx. pipiens mosquito larvae proved to be more susceptible to AuNPs and AgNPs than leaf extract alone by about 6.91 and 6.76 times, respectively.     

Screening and evaluation of different algal extracts and prospects for controlling the disease vector mosquito Culex pipiens L.

Continual application of synthetic insecticides in controlling mosquito larvae has resulted in several problems as build-up of mosquito resistance beside to negative impacts on human health and environment. Discovering new and affordable bio-insecticidal agents with high efficiency, cost effective and target specific become a crucial need. The current study assessed the larvicidal activity of eight methanolic algal extracts belong to three different algal divisions against the 3^rd larval instar of Culex pipiens L. (Diptera: Culicidae). Comparative studies showed that four species of red and green algal extracts exhibited good larvicidal activity. Galaxaura elongata and Jania rubens (Rhodophyta), Codium tomentosum and Ulva intestinales (Chlorophyta) showed higher larvicidal potencies than Padina boryana, Dictyota dichotoma, and Sargassum dentifolium (Phaeophyta) and Gelidium latifolium (Rhodophyta). The maximum level of toxicity was achieved by exposure to G. elongata extract with LC₅₀ (31.13 ppm), followed by C. tomentosum (69.85 ppm) then J. rubens (84.82 ppm) and U. intestinalis (97.54 ppm), while the lowest toxicity exhibited by G. latifolium (297.38 ppm) at 72 h post- treatment. The application of LC₅₀ values of G. elongate, J. rubens, C. tomentosum, and U. intestinalis extracts affected the activities of antioxidant enzymes viz. superoxide dismutase, catalase and glutathione peroxidase as oxidative stress markers. An increase of antioxidant enzymes activities was recorded. Therefore, a significant elimination of free radicals, causing toxic effects. Overall, this study casts light on the insecticidal activity of some algal extracts, suggesting the possibility of application of these bio- agents as novel and cost- effective larvicides.     

Larvicidal activity and Histopathological changes of Cinnamomum burmannii,Syzygium aromaticum extracts and their combination on Culex pipiens

In order to develop an eco-friendly botanical larvicide alternative to the synthetic larvicides, extracts were prepared from the Cinnamomum burmannii (C.B.) and Syzygium aromaticum (S.A.) with hexane using a sonicator. The extracts were evaluated for larvicidal activity individually and in combination against the Culex pipiens larvae. The LC₅₀ value of C.B. and the S.A. hexane extracts tested individually were 184.2 and 363.7 µg/mL against Cx. pipiens respectively. All the combinations of the extract of C.B. and S.A. showed synergistic factors higher than one. Among the different ratios of extracts, the SA25%: CB75% extract was found to be more toxic than the other combinations (LC₅₀:125.7 µg/mL). Midgut cells treated with S.A. 25%: C.B. 75% extract showed severe morphological alterations such as degradation of microvilli; degeneration of epithelial cells, and peritrophic membrane; loss of nuclei, irregular and damage of microvilli. The extract has a promising larvicidal potential against Cx. pipiens, However, the extract was toxic against HUVEC cells, as evident from MTT and cell morphology. Further investigation is required to assess the toxicity of the extract on aquatic animals.     

Chemical composition and larvicidal activity of Piper capense essential oil against the malaria vector,Anopheles gambiae

Hydro-distilled essential oil from Kenyan Piper capense (Piperaceae) was analysed by gas chromatography mass spectrometry (GC–MS) and evaluated for larvicidal activity against the malaria vector, Anopheles gambiae. The oil consisted mainly of sesquiterpene hydrocarbons which accounted for 43.9% of the oil. The major sesquiterpenes were δ-cadinene (16.82%), β-bisabolene (5.65%), and bicyclogermacrene (3.30%). The oil also had appreciable amounts of monoterpene hydrocarbons (30.64%), including β-pinene (7.24%) and α-phellandrene (4.76%), and arylpropanoids (8.64%), including myristicin (4.26%). The oil showed larvicidal activity against third instar larvae of A. gambiae, with LC₅₀ and LC₉₀ values of 34.9 and 85.0 ppm, respectively. Most of the larvae died within the first few hours. The high larvicidal activity of this oil was indicated by the fact that over 80% mortality was observed at a concentration of 100 ppm after 24 h. These results compared favourably with the commercial larvicide pylarvex® which had LC₅₀ and LC₉₀ values of 3.7 and 7.8 ppm, respectively. Application of this oil or of products derived from it to larval habitats may lead to promising results in malaria and mosquito management programmes.     

Larvicidal and mosquito repellent action of peppermint (Mentha piperita) oil

Oil of Mentha piperita L. (Peppermint oil), a widely used essential oil, was evaluated for larvicidal activity against different mosquito species: Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus by exposing IIIrd instar larvae of mosquitoes in enamel trays 6 × 4 inch² size filled to a depth of 3 inch with water. Of the three species tested Cx. quinquefasciatus was most susceptible followed by Ae. aegypti and An. stephensi. Application of oil at 3 ml/m² of water surface area resulted in 100% mortality within 24 h for Cx. quinquefasciatus, 90% for Ae. aegypti and 85% for An. stephensi. For Ae. aegypti 100% mortality was achieved at 3 ml/m² in 48 h or 4 ml/m² in 24 h. For An. stephensi 100% mortality was observed at 4 ml/m² in 72 h. The emergence at 3 ml/m² was also inhibited to a great extent and the few adults which emerged did not ovipost even after taking a blood meal. The oil showed strong repellent action against adult mosquitoes when applied on human skin. Percent protection obtained against An. annularis, An. culicifacies, and Cx. quinquefasciatus was 100%, 92.3% and 84.5%, respectively. The repellent action of Mentha oil was comparable to that of Mylol oil consisting of dibutyl and dimethyl phthalates.     

Insecticidal effects of a novel polyherbal formulation (HF7) against Culex pipiens L. (Diptera: Culicidae)

Plant secondary metabolites represent the most efficient and convenient method to control and overcome environmental pollution and insecticidal resistance. This study explored the mosquitocidal activity of the combined extract of seven plants, (HF7) extracted using a Soxhlet extractor against Culex pipiens under laboratory conditions. Exposure of the 3rd instars of Cx. pipiens to HF7 hexane extract resulted in LC₅₀:114.5 μg/mL and LC₉₀:117.0 μg/mL values after 24 h. The ovicidal activities of hexane extract against Cx. pipiens eggs were 21.6%, 48.3%, and 71.6% at 187.5, 93.7, and 46.88 μg/mL, respectively. HF7-treated larvae showed the formation of irregular blebbing of epithelial cells toward the lumen and sloughing into the gut lumen. HF7 extract resulted in 100% adulticidal mortality at the concentration of 3.7 mg/test tube after 30 min of exposure. The IC₅₀ of HF7 extract was 97.03 µg/ml against larvae, at which nuclear and morphological changes were observed. The spectroscopy spectrum of HF7 hexane extract disclosed the presence of 57 different secondary metabolites, among which the dominant compound was eugenol (32.3%). HF7 hexane extract could serve as a botanical insecticide for controlling Cx. pipiens and potentially other mosquito species.     

Chemical composition and larvicidal effects of essential oil of Dendropanax morbifera against Aedes aegypti L.

Essential oils obtained from the flowers of Dendropanax morbifera were extracted and the chemical composition and larvicidal effects were studied. The analyses were conducted by gas chromatography and mass spectroscopy (GC–MS) revealed that the essential oil of D. morbifera contained 27 compounds. The major chemical components identified were γ-elemene (18.59%), tetramethyltricyclohydrocarbon (10.82%), β-selinene (10.41%), α-zingibirene (10.52%), 2-isopropyl-5-methylbicylodecen (4.2%), β-cubebene (4.19), and 2,6-bis(1,1-Dimethylethyl)-4-phenol (4.01%). The essential oil had a significant toxic effect against early fourth-stage larvae of Aedes aegypti L. with an LC₅₀ value of 62.32 ppm and an LC₉₀ value of 131.21 ppm. The results could be useful in search for newer, safer, and more effective natural larvicidal agents against A. aegypti.     

Toxicity of Acalypha indica (Euphorbiaceae) and Achyranthes aspera (Amaranthaceae) leaf extracts to Aedes aegypti (Diptera: Culicidae)

Alternative control strategies for the dengue vector Aedes aegypti L. (Diptera: Culicidae) include botanical insecticides. They are believed to pose little threat to the environment or to human health and may provide practical substitutes for synthetic insecticides. In this study, we determined the biological activities of methanol extracts of Acalypha indica L. (Euphorbiaceae) and Achyranthes aspera L (Amaranthaceae) leaves individually and in combination as botanical insecticides against Ae. aegypti. Based on LC₅₀ values for 4th instar Ae. aegypti, the combined extracts showed the strongest larvicidal activity (277 ppm). A. aspera and A. indica extracts individually gave similar results (409 and 420 ppm, respectively). Respective LC₅₀ values for pupae were 326 ppm, 456 ppm, and 467 ppm. In studies of smoke toxicity, 64% of females exposed to negative control smoke (no extract) blood fed on chicken, whereas 17% blood fed when exposed to smoke containing A. aspera extract and to positive control smoke (0.2% d-allethrin). In the field, treatment of water storage tanks (≈ 0.5 m³) with combined plant extract reduced larval and pupal populations by 97% and 81%, respectively, after 5 days. Given the results of this study, further evaluation of the combined (A. indica + A. aspera) extract as a mosquito larvicide is warranted. Mosquito coils with A. aspera extract also show promise as a practical and potentially economical means for mitigating mosquito blood feeding.     

Bioassay-guided fractionation of a dried commercial source,Alpinia galanga (L.) Willd rhizomes extract,against Culex pipiens (Diptera: Culicidae)

The increasing risk of insecticide resistance in mosquito populations has led to the search for new larvicidal agents. Evaluation of bioassay-guided fractionation of the rhizome extract of Alpinia galanga (L.) Willd against Aedes aegypti was assessed. Bioactive fractions were isolated from the rhizome extract of A. galanga using a Soxhlet extractor and chromatography techniques, and subsequently tested against the fourth instar of Culex pipiens. The lethal concentration (LC) was calculated via log-probit analysis. The active fraction was evaluated by gas chromatography-mass spectroscopy (GC–MS) and infrared (IR) analysis. The highest larvicidal potential obtained from bioassays using the Soxhlet apparatus was observed in dichloromethane (DCM) and ethyl acetate (EtoAc) extracts, with LC₅₀ values of 124.49 and 176.30 ppm, respectively, after 24 h of exposure. Subsequently, the DCM extract was subjected to column and thin-layer chromatography. Results of the DCM extraction and the active TLC fraction (F133) of the Rf value 0.5 revealed that LC₅₀ and LC₉₀ values decreased over time. The F133 fraction of A. galanga exhibited zero hatchability (100% mortality) at LC₅₀ (63.416 ppm) and LC₂₅ (31.70 ppm) against Cx. pipiens eggs. GC–MS analysis of the active thin-layer chromatography TLC fraction (F133) revealed the presence of phenol 2 4-bis (1 1-dimethylethyl), which was identified as the major compound. Alpinia galanga extract is a promising candidate for the control of mosquito populations. Further study is required to determine the effect of the extracts on non-target organisms.     

Acetylenic 2-phenylethylamides and new isobutylamides from Acmella oleracea (L.) R. K. Jansen,a Brazilian spice with larvicidal activity on Aedes aegypti

Ethanol extract obtained from dried leaves of Acmella oleracea afforded after a liquid/liquid partition procedure a larvicidal hexane fraction (LC₅₀ = 145.6 ppm) and a non larvicidal dichloromethane one. From the inactive fraction, three amides were identified, two new structures, named deca-6,9-dihydroxy-(2E,7E)-dienoic acid isobutylamide (1), deca-8,9-dihydroxy-(2E,6Z)-dienoic acid isobutylamide (2) and the known nona-2,3-dihydroxy-6,8-diynoic acid 2-phenylethylamide (3). Bioassay-guided chromatographic fractionation of the hexane partition led to the identification of an amide mixture, nona-(2Z)-en-6,8-diynoic acid 2-phenylethylamide (4) and deca-(2Z)-en-6,8-diynoic acid 2-phenylethlylamide (5). This mixture was active against Aedes aegypti larvae at LC₅₀ = 7.6 ppm. Low toxicity of crude extracts and derived fractions on Artemia salina nauplies showed the possibility of using them to control the A. aegypti mosquito larvae. This is the first report on larvicidal activity of acetylenic 2-phenylethylamides and their identification in A. oleracea leaves.     

Piperonyl butoxide synergizes the larvicidal activity of Origanum vulgare essential oil and its major constituents against the larvae of Aedes albopictus and Culex pipiens quinquefasciatus

Mosquitoes bite human beings and transmit many diseases, such as malaria, dengue fever, and Zika virus. Vector control of mosquitoes is an effective strategy for reducing the spread of disease. However, extensive use of insecticides (e.g. pyrethroids and organophosphorus) has caused resistance in mosquitoes, which weakens the effectiveness of mosquito control. Phytochemicals have been considered an alternative approach to mosquito control. Essential oil (EO) was obtained from the leaves and flowers of Origanum vulgare, and its synergistic activity with piperonyl butoxide (PBO) was tested against Aedes albopictus and Culex pipiens quinquefasciatus larvae. Thirty-seven compounds were identified, among which carvacrol and thymol were two major constituents (30.73 % and 18.81 %, respectively). O. vulgare EO had a significant toxic effect against fourth-stage larvae of Cx. p. quinquefasciatus and Ae. albopictus, with LC₅₀ values of 17.51 and 75.90 mg/L. Carvacrol and thymol also each appeared to be more effective against Cx. p. quinquefasciatus (LC₅₀ = 19.30 and 11.56 mg/L, respectively) than Ae. albopictus (LC₅₀ = 26.62 and 26.66 mg/L, respectively). PBO interacted synergistically with O. vulgare EO, carvacrol and thymol with 2.60–6.26 times as much of the active compound needed without PBO as against Cx. p. quinquefasciatus and Ae. albopictus larvae. Overall, our results contribute to the development of new natural mosquito insecticides.