Eco-friendly bio-larvicidal and antimicrobial activity of isolated bioactive compound from Kurthia gibsonii (Bacillales).

The targeted organisms include mosquito vectors, bacterial pathogens and non-targeted organisms. Preliminary mosquito larvicidal activity was conducted using cell-free supernatants (CFSs) from 11 gut bacteria. Among them, the bacterium SS11 exhibited promising results and was identified as Kurthia gibsonii based on its 16S rRNA sequence (1350 bp). The diethyl ether extract (DEE) of K. gibsonii demonstrated significant larvicidal effects, with LC50 values of 5.59 µL/mL and 8.59 µL/mL for 3rd instar larvae of Aedes aegypti and 2nd instar larvae of Anopheles stephensi, respectively. Analysis of the DEE using FT-IR, and GC-MS revealed the presence of 16 functional groups, and 7 bioactive compounds, respectively. A molecular docking study identified GC-MS compounds against odorant receptors from A. aegypti and odorant-binding proteins from A. stephensi was performed to assess the interaction and binding affinity. Overall, these findings suggest that the bioactive compounds 2, 4, 6-tribromoaniline from the DEE of K. gibsonii hold potential as an environmentally compatible alternative for biocontrol purposes, and compounds 9-tricosene and didecyl phthalate can be used for mosquito traps.

Overexpression of cytochrome P450 and esterase genes involved in permethrin resistance in larvae and adults of Culex quinquefasciatus.

Mosquitoes are important vectors of several arthropod-borne diseases, which remain a priority for epidemiological research. Mosquito vector control strategies have traditionally relied on chemical insecticides such as synthetic pyrethroids. However, the indiscriminate use of pesticides has resulted in the development of resistance in many mosquito species. In insects, resistance evolves primarily through the overexpression of one or more gene products from the cytochrome P450, carboxylesterase, and glutathione superfamilies. The current study examined the expression of cytochrome P450 CYP6M2, CYP6AA7, CYP6Z2, CYP9J34, α-Esterase, Esterase B1, and neuroactin genes in larvae and adults of a permethrin-resistant (PerRes) and susceptible (Sus) Culex quinquefasciatus strains. The results showed that the CYP6AA7 gene was overexpressed (10-fold) in larvae and adults with PerRes (p < 0.01) followed by CYPJ34 (9.0-fold) and CYP6Z2 (5.0-fold) compared to the Sus, whereas fewer changes in CYP6M gene expression were observed in PerRes adults (p < 0.05), and no expression was found in larvae. The esterase gene was overexpressed in PerRes larvae (9.0-fold) followed by adults (2.5-fold) compared to the susceptible strain. Based on data, the present study suggests that cytochrome P450, CYP6AA7, CYP6Z2, CYP9J34, α-Esterase, Esterase B1, and neuroactin genes were involved in permethrin resistance in larval and adult Cx. quinquefasciatus.

Effect of melatonin and luzindole antagonist on fipronil toxicity, detoxification and antioxidant enzyme system in different tissues of Helicoverpa armigera (Lepidoptera: Noctuidae).

Studies have investigating the detoxification and antioxidant enzymes with melatonin under pesticide stress in many vertebrates, whereas no reports produced in invertebrates. In this study possible role of melatonin and luzindole effect on fipronil toxicity and the detoxification, antioxidant enzymes in H. armigera has been reported. Result showed high toxicity of fipronil treatment (LC50 4.24 ppm), followed by increased LC50 value with melatonin pretreatment (6.44 ppm). Whereas decreased toxicity was observed with melatonin and luzindole combination (3.72 ppm). The detoxification enzymes AChE, esterase and P450 were increased in larval head and whole body with exogenous melatonin level compared to control 1-1.5 µmol/mg of protein. The antioxidant levels of CAT, SOD and GST in whole body and head tissue had been increased by melatonin and fipronil combination 1.1-1.4 unit/mg of protein followed by GPx and GR in larval head (1-1.2 µmol/mg of protein). Mean while the luzindole antagonist inhibits CAT, SOD, GST and GR oxidative enzyme level (1-1.5 fold) in most of the tissue compared to melatonin and fipronil treatment (p < 0.01). Hence this study concludes that the melatonin pretreatment can reduce the fipronil toxicity by enhanced detoxification and antioxidant enzyme system in H. armigera.

Impact assessment of natural variations in different weather factors on the incidence of whitefly, Bemisia tabaci Genn. and yellow vein mosaic disease in Abelmoschus esculentus (L.) Moench.

Bemisia tabaci Gennadius, also renowned as the silver leaf whitefly, is among the most damaging polyphagous insect pests in many commercially important crops and commodities. A set of field experiments were conducted for three consecutive years i.e., 2018-2020, to investigate the role of variations in rainfall, temperature, and relative humidity on the abundance of B. tabaci in okra (Abelmoschus esculentus L. Moench). In the first experiment, the variety Arka Anamika was cultivated twice a year to analyse the incidence of B. tabaci concerning the prevailing weather factors and the overall pooled incidence recorded during the dry and wet season was 1.34 ± 0.51 to 20.03 ± 1.42 and 2.26 ± 1.08 to 18.3 ± 1.96, respectively. Similarly, it was observed that the highest number of B. tabaci catch (19.51 ± 1.64 whiteflies/3 leaves) was recorded in morning hours between 08:31 to 09:30 a.m. The Yellow Vein Mosaic Disease (YVMD) is a devastating disease of okra caused by begomovirus, for which B. tabaci acts as a vector. In another experiment, screening was conducted to check the relative susceptibility of three different varieties viz., ArkaAnamika, PusaSawani, and ParbhaniKranti against B. tabaci (incidence) and YVMD ((Percent Disease Incidence (PDI), Disease Severity Index (DSI), and Area Under the Disease Progress Curve (AUDPC)). The recorded data was normalized by standard transformation and subjected to ANOVA for population dynamics and PDI. Pearson's rank correlation matrix and Principal Component Analysis (PCA) have been used to relate the influences of various weather conditions on distribution and abundance. SPSS and R software were used to create the regression model for predicting the population of B. tabaci. Late sown PusaSawani evolved as a highly susceptible variety in terms of B. tabaci (24.83 ± 6.79 adults/3leaves; mean ± SE; N = 10) as well as YVMD i.e., PDI (38.00 ± 4.95 infected plants/50plants), DSI (71.6-96.4% at 30 DAS) and AUDPC (mean ß-value = 0.76; R2 = 0.96) while early sown Parbhani Kranti least susceptible to both. However, the variety ArkaAnamika was observed as moderately susceptible to B. tabaci and its resultant disease. Moreover, environmental factors were predominantly responsible for regulating the population of insect pests in the field and hence its productivity like rainfall and relative humidity were negative while the temperature was positively correlated with B. tabaci (incidence) and YVMD (AUDPC). The findings are helpful for the farmers to choose need-based IPM strategies than timing-based, which would fit perfectly with the present agro-ecosystems in all ways.

Synergism and toxicity of iron nanoparticles derived from Trigonella foenum-graecum against pyrethriod treatment in S. litura and H. armigera (Lepidoptera: Noctuidae).

The tobacco cutworm, Spodoptera litura and cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae) are important pests of various agricultural crops that cause sevier economic loses throughout the world. Indiscriminate and frequent use of insecticide may lead to development of resistance in these pests. Nanotechnology has given an alternative to manage and overcome insecticide resistance for pest management strategies. In the present study the iron nanoparticles derived from Trigonella foenum-graecum leaf extract (FeNPs) was investigated for its ecofriendly management of pyrethroid resistance in two lepidopteron pest species at 24 h, 48 h and 72 h post treatment. The result showed high mortality (92.83% and 91.41%) of S. litura and H. armigera at 72 h treatment upon FeNPs and fenvalerate (Fen + FeNPs) teratment. Probit analysis revealed high LC50 upon Fen + FeNPs treatment (130.31 and 89.32 mg/L) with a synergism ratio of 1.38 and 1.36. Antifeedant activity of six dofferent concentration of FeNPs revelaed increased antifeedant activity with respect to increasing concentration of nanoparticles ranging from 10 to 90% and 20-95% againt both insects (p<0.05). Detoxification activity of carboxylesterase was elevated at 630 µmol/mg protein/min (p<0.05) in fenvalerate treatment, whereas decreased activity was found (392umole/mg protein/min) in FeNPs and Fen + FeNPs treatment (P<0.001). GST and P450 activity was also increased in fenvalerate treatment, whereas decreased activity was observed in FeNPs and Fen + FeNPs. Esterase isoenzyme banding pattern revealed four bands in fenvalerate treatment and two bans (E3 and E4) in Fen + FeNPs combination. Hence the present study concludes that T. foenum-graecum synthesized iron nanoparticles could be an effective alternate for ecofriendly management of S. litura and H. armigera.

Effects of phytocompound Precocene 1 on the expression and functionality of the P450 gene in λ-cyhalothrin-resistant Spodoptera litura (Fab.).

Spodoptera litura (Fabricius) is an agriculturally significant polyphagous insect pest that has evolved a high level of resistance to conventional insecticides. A dietary assay was used in this work to assess the resilience of field populations of S. litura to λ-cyhalothrin. Analysis of the function and expression of the cytochrome P450 gene was used to test the sensitivity of S. litura larvae to sub-lethal concentrations of the insecticidal plant chemical Precocene 1, both by itself and in combination with λ-cyhalothrin. The activity of esterase enzymes (α and ß) was found to decrease 48 h post treatment with Precocene 1. The activity of GST enzyme and cytochrome P450 increased with Precocene 1 treatment post 48 h, however. Expression studies revealed the modulation by Precocene 1 of cytochrome P450 genes, CYP4M16, CYP4M15, CYP4S8V4, CYP4G31, and CYP4L10. While CYP4M16 expression was stimulated the most by the synergistic Precocene 1 + λ-cyhalothrin treatment, expression of CYP4G31 was the most down-regulated by Precocene 1 exposure. Hence, it is evident that λ-cyhalothrin-resistant pest populations are still sensitive to Precocene 1 at a sublethal concentration that is nevertheless capable of hindering their development. Precocene 1 can therefore be considered a potent candidate for the effective management of insecticide-resilient S. litura.

Pretreatment of mosquito larvae with ultraviolet-B and nitropolycyclic aromatic hydrocarbons induces increased sensitivity to permethrin toxicity.

Nitropolycyclic aromatic hydrocarbons (Nitro-PAH) are highly toxic PHA derivatives. Nitro-PHAs are emitted by carbonaceous materials and PHA post-emission transformation, which causes water and environmental pollution and also exists as carcinogenic and immunotoxic agents. UV light has been shown to cause DNA damage and improves the covalent binding of PAH to DNA significantly. Mosquito breeding grounds are pools of water that can be large open zones or encased ponds with varying levels of sunlight exposure. This research was performed to assess the combined effects of UV-B exposure and Nitro-PAH on the physiological function of Culex quinquefasciatus larvae. To assess the impact of UV-B irradiation and Nitro-PAH exposure on mosquito vectors, parameters were examined: (1) Nitro-PAH availability and its impact on cell fatalities; (2) the detoxifying abilities of cytochrome P450, glutathione-S-transferase, and esterase; (3) the reactions to Reactive Oxygen Species; and (4) The resistance of mosquito larvae to three synthetic pesticides (temephos, imidacloprid, and permethrin). UV-B and Nitro-PAH treatment caused cellular damage and increased major detoxification enzymes such as α & ß-esterase, cytoP450, CAT, GST, and POX. The levels of oxidative stress, ROS and protein carbonyl content, nitrite, ascorbic acid and thiobarbituric acid were decreased significantly. Toxicology bioassays revealed that UV-B + Nitro-PAH exposure significantly increased larval susceptibility. The current study concludes that prior exposure to Nitro-PAHs and UV-B may make mosquito larvae more vulnerable to chemical insecticides.

Green synthesis of gold nanoparticles using Gracilaria crassa leaf extract and their ecotoxicological potential: Issues to be considered.

The use of vegetal species for gold nanoparticles (AuNPs) biosynthesis can constitute an alternative to replacing the extensive use of several hazardous chemicals commonly used during NPs synthesis and, therefore, can reduce biological impacts induced by the release of these products into the natural environment. However, the "green nanoparticles" and/or "eco-friendly nanoparticles" label does not ensure that biosynthesized NPs are harmless to non-target organisms. Thus, we aimed to synthesize AuNPs from seaweed Gracilaria crassa aqueous extract through an eco-friendly, fast, one-pot synthetic route. The formation of spherical, stable, polycrystalline NPs with a diameter of 32.0 nm ± 4.0 nm (mean ±SEM) was demonstrated by UV-vis spectroscopy, field emission scanning electron microscopy, and high-resolution transmission electron microscopy, energy-dispersive X-ray and X-ray diffraction measurement, and Fourier-transform infrared spectroscopy analysis. In addition, different phytocomponents were identified in the biosynthesized AuNPs, using Gas Chromatography-Mass Spectrometry (GC-MS). However, both G. crassa aqueous extract and the biosynthesized AuNPs showed high ecotoxicity in Anopheles stephensi larvae exposed to different concentrations. Therefore, our study supports the potential of seaweed G. crassa as a raw material source for AuNPs biosynthesis while also shedding light on its ecotoxicological potential, which necessitates consideration of its risk to aquatic biota.

Characterization and Evaluation of Metarhizium spp. (Metsch.) Sorokin Isolates for Their Temperature Tolerance.

A field survey was done in teak (Tectona grandis F.) forests in South India to explore the entomopathogenic effect of Metarhizium anisopliae (Ascomycota: Sordariomycetes) against teak defoliator, Hyblaea puera (Lepidoptera: Hyblaeidae). About 300 soils and infected insect samples were collected during the survey and thirty-six fungal isolates were isolated from soil and insect samples and characterized. The fungi were cultured on PDAY with dodine and antibiotics. Generally, the EPF culture was incubated at 27 °C in darkness for 15 days. Virulence of the Entomopathogenic Fungi (EPF) ability to germinate under cold and heat temperatures was assessed in a culture impregnated with conidia. In the experiment, it was found that for the first time Metarhizium quizhouense, Metarhizium robertsii, and Metarhizium majus species caused significantly higher mortality to hosts. These isolates of M. anisopliae, M. robertsii, M. majus, and M. quizhouense were all considered to be effective virulent and environmentally adaptive. The Metarhizium isolates were recommended as effective bio-control agents through the field investigation of teak defoliator Hyblaea puera from South India forest. This study paves the way to utilize the indigenous isolates of EPF for the control of teak defoliator and to combat the pests thatare resistant to insecticide.

RNA Interference Suppression of v-ATPase B and Juvenile Hormone Binding Protein Genes Through Topically Applied dsRNA on Tomato Leaves: Developing Biopesticides to Control the South American Pinworm, Tuta absoluta (Lepidoptera: Gelechiidae).

The South American pinworm Tuta absoluta (Meyrick) (Family: Gelechiidae) is one of the most devastating lepidopteran pests in the developing countries of South America, Africa, and Asia. This pest is classified as the most serious threat for tomato production worldwide. In the present study, we analyzed RNAi-mediated control through exogenously applied dsRNA delivery on tomato. The dsRNA treatments were made to target the juvenile hormone binding protein and the v-ATPase B. Both mRNA targets were cloned, validated by sequencing, and used to produce each dsRNA. After treatments the relative transcript expression was analyzed using qRTPCR to assess to efficacy of RNAi. A leaf-dip assay was used to provide late 2nd instar larvae three feeding access periods: 24, 48, and 72 h, to evaluate the effect of gene silencing of each target. Larvae were fed tomato leaves coated with five different RNAi concentrations (10, 20, 30, 40, and 50 micrograms/centimeter-squared), that suppressed two genes (juvenile hormone protein, JHBP, and vacuolar-type adenosine triphosphatase enzyme, v-ATPase). Treatments with dsRNA showed a significant increase in mortality at 24, 48, and 72 h after ingestion (P < 0.01, α = 0.05), along with reduced leaf damage, and increased feeding deterrence. The results suggest that these two RNAi products may provide a suitable treatment for control of this and other lepidopteran pests.

Efficacy of Precocene I from Desmosstachya bipinnata as an Effective Bioactive Molecules against the Spodoptera litura Fab. and Its Impact on Eisenia fetida Savigny.

The sustainability of agroecosystems are maintained with agro-chemicals. However, after more than 80 years of intensive use, many pests and pathogens have developed resistance to the currently used chemistries. Thus, we explored the isolation and bioactivity of a chemical compound, Precocene I, isolated from the perennial grass, Desmosstachya bipinnata (L.) Stapf. Fractions produced from chloroform extractions showed suppressive activity on larvae of Spodoptera litura (Lepidoptera: Noctuidae), the Oriental armyworm. Column chromatography analyses identified Precocene I confirmed using FTIR, HPLC and NMR techniques. The bioactivity of the plant-extracted Dp-Precocene I was compared to a commercially produced Precocene I standard. The percentage of mortality observed in insects fed on plant tissue treated with 60 ppm Db-Precocene I was 97, 87 and 81, respectively, for the second, third and fourth instar larvae. The LC50 value of third instars was 23.2 ppm. The percentages of survival, pupation, fecundity and egg hatch were altered at sub-lethal concentrations of Db-Precocene I (2, 4, 6 and 8 ppm, sprays on castor leaves). The observed effects were negatively correlated with concentration, with a decrease in effects as concentrations increased. Distinct changes in feeding activity and damage to gut tissues were observed upon histological examination of S. litura larvae after the ingestion of Db-Precocene I treatments. Comparative analyses of mortality on a non-target organism, the earthworm, Eisenia fetida, at equal concentrations of Precocene I and two chemical pesticides (cypermethrin and monocrotophos) produced mortality only with the chemical pesticide treatments. These results of Db-Precocene I as a highly active bioactive compound support further research to develop production from the grass D. bipinnata as an affordable resource for Precocene-I-based insecticides.

Genome mining and functional analysis of cytochrome P450 genes involved in insecticide resistance in Leucinodes orbonalis (Lepidoptera: Crambidae).

Genome-wide analysis of cytochrome P450 monooxygenase (CYP) genes from the advanced genome project of the Leucinodes orbonalis and the expression analysis provided significant information about the metabolism-mediated insecticide resistance. A total of 72 putative CYP genes were identified from the genome and transcriptome of L. orbonalis. The genes were classified under 30 families and 46 subfamilies based on the standard nomenclature. In the present study, a novel CYP gene, CYP324F1, was identified and it has not been reported from any other living system so far. Biochemical assays showed enhanced titers (5.81-18.5-fold) of O-demethylase of CYP in five field-collected populations. We selected 34 homologous CYP gene sequences, seemed to be involved in insecticide resistance for primer design and quantitative real-time PCR studies. Among the many overexpressed genes (>10 fold), the expression levels of CYP324F1 and CYP306A1 were prominent across all the field populations as compared with the susceptible iso-female line. Oral delivery of ds-CYP324F1 and ds-CYP306A1 directed against CYP324F1 and CYP306A1 to the larvae of one of the insecticide resistance populations caused reduced expression of these two transcripts in a dose-dependent manner (53.4%-85.0%). It appears that the increased titer of O-demethylase is the result of increased transcription level of CYP genes in resistant populations. The data provide insight for identifying the novel resistance management strategies against L. orbonalis.

Culex quinquefasciatus Egg Membrane Alteration and Ovicidal Activity of Cipadessa baccifera (Roth) Plant Extracts Compared to Synthetic Insect Growth Regulators.

BACKGROUND: Insecticide resistance among mosquito vectors for synthetic insecticides still remains a major problem for control efforts. This study assessed the ovicidal potential of crude solvent extracts from the medicinal plant Cipadessa baccifera comparatively to standard registered synthetic insect growth regulators (IGR) on freshly laid eggs of Culex quinquefasciatus. METHOD: Five plant extracts were prepared using different solvents. The batches of eggs were exposed to different concentrations of each solvent extract comparatively to synthetic IGR. The hatched eggs of Cx. quinquefasciatus were subjected to different concentrations. The first instars that emerged from the eggs were counted daily. The egg hatching inhibition was observed 24, 48 and 72 hrs post treatment. The desiccation median time (DT50 and DT90) was calculated. RESULTS: The percent egg hatching inhibition was inversely proportional to the concentration of extracts. The morphological damage to the eggs was observed. Among five solvent extracts, acetone extracts showed the highest ovicidal activity. The changes in eggshell morphology were observed. The maximum ovicidal activity was observed in acetone extracts with DT50 value of 1.70 hrs (0.91-2.22). The methanol plant extract using gas chromatography-mass spectrometry identified 14 compounds. CONCLUSION: These results suggest that the acetone extracts of C. baccifera have the potential to be used as an ovicidal agent for controlling mosquito populations in aquatic stages. The biodegradability of the extracts has the advantage of being eco-friendly.

Comparative Analysis of Major Mosquito Vectors Response to Seed-Derived Essential Oil and Seed Pod-Derived Extract from Acacia nilotica.

Botanical metabolites are increasingly realized as potential replacements to chemical insecticides. In the present study, Acacia nilotica seed essential oil and seed pod solvent extracts were tested for bioefficacy against three important types of mosquitoes. Mortality was recorded 24 h post-treatment, while smoke toxicity of adult mosquitoes was recorded at 10 min intervals for 40 min. Seed pod powder was extracted with different solvents and hydrodistilled seed oil chemical constituents were determined by using Gas chromatography mass spectroscopy (GC-MS) -. Larvicidal and adulticidal efficacy of seed hydrodistilled essential oil and solvent extracts were tested against larval and adult mosquitoes. The seed hydrodistilled oil provided strong larvicidal activity against Anopheles stephensi, (LC50 (lethal concentration that kills 50% of the exposed larvae) = 5.239, LC90 (lethal concentration that kills 90% of the exposed larvae) = 9.713 mg/L); Aedes aegypti, (LC50 = 3.174, LC90 = 11.739 mg/L); and Culex quinquefasciatus, (LC50 = 4.112, LC90 = 12.325 mg/L). Smoke toxicities were 82% in Cx. quinquefasciatus, 90% in Ae. aegypti, and 80% mortality in An. stephensi adults, whereas 100% mortality was recorded for commercial mosquito coil. The GC-MS profile of seed essential oil from A. nilotica showed the presence of hexadecane (18.440%) and heptacosane (15.914%), which are the main and active compounds, and which may be involved in insecticidal activity. Overall findings suggest that the seed oil showed strong mosquitocidal activity against mosquito vectors and therefore may provide an ecofriendly replacement to chemical insecticides.

Mosquitocidal Effect of Glycosmis pentaphylla Leaf Extracts against Three Mosquito Species (Diptera: Culicidae).

BACKGROUND: The resistance status of malaria vectors to different classes of insecticides used for public health has raised concern for vector control programmes. Alternative compounds to supplement the existing tools are important to be searched to overcome the existing resistance and persistence of pesticides in vectors and the environment respectively. The mosquitocidal effects of Glycosmis pentaphylla using different solvents of acetone, methanol, chloroform and ethyl acetate extracts against three medically important mosquito vectors was conducted. METHODS: Glycosmis pentaphylla plant leaves were collected from Kolli Hills, India. The WHO test procedures for larval and adult bioassays were used to evaluate extracts against mosquito vectors, and the chemical composition of extracts identified using GC-MS analysis. RESULTS: The larvicidal and adulticidal activity of G. pentaphylla plant extracts clearly impacted the three species of major mosquitoes vectors. Acetone extracts had the highest larvicidal effect against An. stephensi, Cx. quinquefasciatus and Ae. aegypti with the LC50 and LC90 values of 0.0004, 138.54; 0.2669, 73.7413 and 0.0585, 303.746 mg/ml, respectively. The LC50 and LC90 adulticide values of G. pentaphylla leaf extracts in acetone, methanol, chloroform and ethyl acetate, solvents were as follows for Cx. quinquefasciatus, An. stephensi and Ae. Aegypti: 2.957, 5.458, 2.708, and 4.777, 3.449, 6.676 mg/ml respectively. The chemical composition of G. pentaphylla leaf extract has been found in 20 active compounds. CONCLUSIONS: The plant leaf extracts of G. pentaphylla bioactive molecules which are effective and can be developed as an eco-friendly approach for larvicides and adulticidal mosquitoes vector control. Detailed identification and characterization of mosquitocidal effect of individual bioactive molecules ingredient may result into biodegradable effective tools for the control of mosquito vectors.

Insecticidal potential of Ocimum canum plant extracts against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus larval and adult mosquitoes (Diptera: Culicidae).

Mosquitoes have developed resistance to various synthetic insecticides, making their control increasingly difficult. Insecticides of botanical origin may serve as suitable natural control. This study evaluates the toxic potential of Ocimum canum (Sims) leaf extract and powder against Anopheles stephensi (Liston), Aedes aegypti (Lin) and Culex quinquefasciatus (Say) larval and adult mosquitoes. Larval mortality was observed after 24 h recovery period and adult smoke toxicity observed for 40 min duration at 10 min interval. Methanol extract of O. canum showed highest larval mortality against the larvae of C. quinquefasciatus LC50 = 28.3225, LC90 = 44.1150; Ae. aegypti LC50 = 43.327, LC90 = 61.249; and An. stephensi LC50 = 30.2001, LC90 = 48.2866 ppm. The smoke toxicities were 93% mortality in C. quinquefasciatus, 74% in Ae. aegypti and 79% in An. stephensi adults, respectively, whereas 100% mortality was recorded in the commercial mosquito control. Our results suggest that O. canum leaf extract and powder are natural insecticide, and ideal eco friendly approach for mosquito control.

Laboratory development of permethrin resistance and cross-resistance pattern of Culex quinquefasciatus to other insecticides.

Resistance of mosquitoes to insecticides is a growing concern in India. Since only a few insecticides are used for public health and limited development of new molecules is expected in the next decade, maintaining the efficacy of control programs mostly relies on resistance management strategies. Developing such strategies requires a deep understanding of factors influencing resistance together with characterizing the mechanisms involved. Among factors likely to influence insecticide resistance in mosquitoes, agriculture and urbanization have been implicated but rarely studied in detail. In the present study, we evaluate the permethrin resistance and cross-resistance pattern of several insecticides in Culex quinquefasciatus mosquitoes. After 10 generation of selection with permethrin, the LC50 value for both larvae and adult Cx. quinquefasciatus was increased by 17.3- and 17.1-folds compared with susceptible strain. Detoxification enzyme profiles and native PAGE electrophoresis of esterase isoenzyme further revealed that esterase and CytP450 may be involved in permethrin resistance (PerRes) strain compared with susceptible strain. In addition to cross-resistance, study revealed that high resistance to cypermethrin (RR = 6.3, 8.8-folds). This study provided important information for understanding permethrin resistance and facilitating a better strategy for the management of resistance. These studies conclude that a strong foundation for further study of permethrin resistance mechanisms observed in Cx. quinquefasciatus mosquitoes.

Insecticidal and repellent activity of Clausena dentata (Rutaceae) plant extracts against Aedes aegypti and Culex quinquefasciatus mosquitoes (Diptera: Culicidae).

Mosquito control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol agents. The present study is to evaluate adulticidal activity of Clausena dentata plant extract against Aedes aegypti and Culex quinquefasciatus mosquitoes. The adult mortality was observed after 24 h of exposure. The highest mortality was found in acetone extracts against Ae. aegypti and Cx. quinquefasciatus with the LC50 and LC90 4.1783 mg/ml (3.8201-7.1026), 9.3884 mg/ml (7. 8258-13.1820) and 4.2451 mg/ml (3.8547-8.0254), 12.3214 mg/ml (10.9287-16.2220), respectively. Smoke toxicity was observed at 10-min interval for 40 min, and the mortality data were recorded. Result shows that Ae. aegypti and Cx. quinquefasciatus are 85 ± 2 and 89 ± 1.5, respectively. A mortality of 100 % was recorded in the commercial mosquito control. These results suggest that the leaf extracts of C. dentata have a potential to be used as an ideal eco-friendly approach for the control of mosquitoes.

Adulticidal and smoke toxicity of Cipadessa baccifera (Roth) plant extracts against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus.

Mosquito vectors are responsible for the transmission of parasitic and viral infections, including loss in commercial and labor outputs, particularly in developing countries with tropical and subtropical climates. The aim of the present study is to evaluate the adulticidal and smoke toxicity of Cipadessa baccifera (Roth) against three important mosquitoes vectors, Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). Adult mortality was observed after 24-h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in acetone extract against An. stephensi followed by Ae. aegypti and Cx. quinquefasciatus with the LD50 and LD90 values 16.021 (14.080-18.345), 29.095 (25.118-34.089); 23.581 (22.100-28.315), 38.636 (35.321-41.021); and 13.560 (9.479-17.391), 248.35 (203.47-344.43) mg/ml, respectively. No mortality was recorded in the control. Smoke toxicity was observed at 10-min interval for 40 min and the mortality data were recorded. Among the C. baccifera plant powder tested. Smoke toxicity results show that Cx. quinquefasciatus, An. stephensi, and Ae. aegypti shows 88.6 ± 1.8, 78.2 ± 0.5, and 77 ± 1, respectively. One hundred percent mortality was recorded in the commercial mosquito control. The present study shows that C. baccifera leaf powder can be used as an efficient toxicity against mosquitoes. These results suggest that the leaf extracts of C. baccifera have a potential to be used as an ideal eco-friendly approach for the control of mosquitoes.

Chemical composition and larvicidal activity of plant extracts from Clausena dentata (Willd) (Rutaceae) against dengue, malaria, and filariasis vectors.

Mosquitoes in the larval stage are attractive targets for pesticides because mosquitoes breed in water, and thus, it is easy to deal with them in this habitat. The use of conventional pesticides in the water sources, however, introduces many risks to people and/or the environment. Natural pesticides, especially those derived from plants, are more promising in this aspect. Aromatic plants and their essential oils are very important sources of many compounds that are used in different respects. Insecticides of botanical origin may serve as suitable alternative to chemical insecticides. Acetone, chloroform, ethyl acetate, methanol, and petroleum benzine leaf extracts of Clausena dentata were tested against the fourth instar larvae of Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti (Diptera: Culicidae). Larval mortality was observed after 24 h of exposure. The highest larval mortality was found in acetone leaf extract, C. quinquefasciatus (LC50 = 0.150278 mg/ml; LC90 = 7.302613 mg/ml), A. aegypti (LC50 = 0.169495 mg/ml; LC90 = 1.10034 mg/ml), and A. stephensi (LC50 = 0.045684 mg/ml; LC90 = 0.045684 mg/ml). GC-MS analysis of plant extracts of acetone solvent revealed 16 compounds, of which the major compounds were benzene,1,2,3-trimethoxy-5-(2-propenyl) (14.97%), Z,Z-6,28-heptatriactontadien-2-one (6.81%), 2-allyl-4-methylphenol (28.14%), 2-allyl-4-methylphenol (17.34%), and 2,6,10,14,18,22-tetracosahexaene, 2,6,10,15,19,23-hexamethyl (10.35%). Our result shows acetone leaf extracts of C. dentata have the potential to be used as an ideal eco-friendly approach for mosquito control.