Insect feeding deterrent and growth inhibitory activities of scopoletin isolated from Artemisia annua against Spilarctia obliqua (Lepidoptera: Noctuidae)

Abstract Artemisia annua (Asteraceae) is well known for its antimalarial activities due to presence of the compound artemisinin. We isolated a methoxy coumarin from the stem part of A. annua and confirmed its identity as scopoletin through mass spectral data. The structure was established from ¹H‐nuclear magnetic resonance (NMR), ¹³C‐NMR. The compound scopoletin was evaluated for its feeding deterrence and growth inhibitory potential against a noxious lepidopteran insect, Spilartctia obliqua Walker. Scopoletin gave FD₅₀ (feeding deterrence of 50%) value of 96.7 μg/g diet when mixed into artificial diet. S. obliqua larvae (12‐day‐old) exposed to the highest concentration (250 μg/g diet) of scopoletin showed 77.1% feeding‐deterrence. In a growth inhibitory assay, scopoletin provided 116.9% growth inhibition at the highest dose of 250 μg/g diet with a GI₅₀ (growth inhibition of 50%) value of 20.9 μg/g diet. Statistical analysis showed a concentration‐dependent dose response relationship toward both feeding deterrent and growth inhibitory activities. Artemisinin is found mainly in the leaves of A. annua and not in the stems, which are typically discarded as waste. Therefore identification of scopoletin in stems of A. annua may be important as a source of this material for pest control.     

Bioefficacy of Ceasalpinea bonduc (L.) Roxb. against Spodoptera litura Fab. (Lepidoptera: Noctuidae)

Bioefficacy of hexane, chloroform and ethyl acetate extracts of the seeds of Ceasalpinea bonduc (L.) Roxb. was studied against third instar larvae of Spodoptera litura at 0.25, 0.5, 1, 2.5 and 5.0% concentrations. Significant antifeedant, larvicidal and pupicidal activities and least LC₅₀ values were observed in chloroform extract. The chloroform extract was subjected to fractionation using silica gel column chromatography. Six fractions were obtained; among these, the third fraction showed high antifeedant, larvicidal and pupicidal activities at 1000 ppm concentration. Abnormalities in adults were also observed. All the activities were concentration dependent. C. bonduc could be useful in integrated pest management programme.     

Effect of ellagic acid on the larvae of Spodoptera litura (Lepidoptera:Noctuidae) and its parasitoid Bracon hebetor (Hymenoptera:Braconidae)

The present study was aimed at investigating the effect of plant based compound, ellagic acid on parasitoid Bracon hebetor (Say) through its host, the common cutworm, Spodoptera litura (Fabricius). The effect on S. litura was ascertained by feeding six days old larvae on artificial diet incorporated with different concentrations (5 ppm, 25 ppm, 125 ppm, 625 ppm, 3125 ppm) of ellagic acid and water as control. Its effect on B. hebetor was determined by allowing the adult B. hebetor to parasitize the treated host larvae. The mortality of S. litura larvae was increased whereas adult emergence declined with increasing concentration of ellagic acid. The developmental period was delayed significantly and all the nutritional indices were reduced with treatment. Ellagic acid at LC₃₀ (7.70 ppm) had not much influence on the growth of parasitoid B. hebetor but LC₅₀ (43.45 ppm) adversely influenced the development of the parasitoid, B. hebetor when reared on treated larvae of S. litura. This was evident from reduced parasitization, fecundity, egg laying, egg hatching, emergence, increased larval mortality, reduced pupation and prolonged development of the immature stages at LC₅₀. However, parasitization, egg hatching and larval mortality of the parasitoid were not significantly impacted at LC₃₀ indicating the possibility of its use in integrated pest management programmes.     

Effect of plant secondary metabolites on common cutworm,Spodoptera litura (Lepidoptera: Noctuidae)

The effect of various flavonoids, lectins and phenyl β‐_D‐glucoside on larval survival, weights and the activities of digestive (total serine protease and trypsin) and detoxifying (esterase and glutathione‐S‐transferase) enzymes of Spodoptera litura larvae at 7 days after treatment was studied through diet incorporation assay. Flavonoids (rutin, chlorogenic acid, quinic acid, caffeic acid, naringenin, quercitin, kaempferol, myricetin, catechin, and ferulic acid) were incorporated in artificial diet at 100, 500 and 1000 ppm, lectins: groundnut leaf lectin (GLL), concavalin A (ConA) and phenyl β‐_D‐glucoside at 1, 2 and 5 μg/mL. Flavonoids such as rutin, quercitin and kaempferol at 1000 ppm were more toxic to S. litura larvae than quinic acid, caffeic acid, naringenin, myricetin, catechin, and ferulic acid. Larval growth and development were significantly reduced in S. litura larvae fed on a diet with GLL and ConA at 5 μg/mL compared to the larvae fed at 2 and 1 μg/mL concentrations. The larvae fed on flavonoid‐treated diets showed significant reduction in serine protease, trypsin and esterase activities. The flavonoids such as rutin, chlorogenic acid, quinic acid, naringenin, quercitin, kaempferol and myricetin, and lectins, GLL and ConA can be utilized in insect control programs.     

Detrimental effects of endophytic fungus Nigrospora sp. on survival and development of Spodoptera litura

Endophytes have been known to confer resistance to host plants against insect herbivores mediated by fungal alkaloids. In this study we have isolated an endophytic fungus Nigrospora sp. (Ascomycota: Sordariomycetes) from a native plant Tinospora cordifolia. To assess anti-insect potential, we tested to what extent the survival and development of Spodoptera litura (Lepidoptera: Noctuidae), a polyphagous pest, was affected when fed on artificial diet amended with different concentrations of ethyl acetate extract of Nigrospora sp. In feeding assay, the insect suffered significantly higher mortality and showed prolonged development period on amended diet as compared to unamended diet. A significant reduction in relative growth and consumption rate, efficiency of conversion of ingested and digested food and approximate digestibility indicated deterrent as well as toxic effects of the fungal extract. Significant adverse effects were also observed on adult emergence, longevity and reproductive potential of S. litura. This preliminary information on insecticidal properties of Nigrospora sp. may further be used for imparting resistance in plants against insects.     

Bacterial compounds,as biocontrol agent against early blight (Alternaria solani) and tobacco cut worm (Spodoptera litura Fab.) of tomato (Lycopersicon esculentum Mill.)

Abstract

Tomato, the important vegetable crop faces yield loses by foliar disease, Early Blight caused by Alternaria solani and infestation by tobacco cut worm, Spodoptera litura. Synthetic pesticides used for disease and pest control resulted in environmental pollution as well as development of resistance. By way of a need to develop a new biopesticide, bacteria were tested for their anti-fungal and insecticidal activity. Volatile compounds and peptides from the bacteria, Bacillus subtilis, Staphylococcus aureus and Pseudomonas aeruginosa were able to inhibit the hyphal growth and melanin production of A. solani. S. aureus showed the highest inhibitory effect as well as reduced the disease severity. B. subtilis exhibited the highest mortality rate of 87% and 83% against I and II instar of S. litura, respectively. The bacteria offered effective biocontrol at 35?°C. Bioactive substances from the bacteria can be used as potential biocontrol agents against the pest and disease of tomato.     

Synthesis,in vitro antitumour activity,and molecular docking study of novel 2-substituted mercapto-3-(3,4,5-trimethoxybenzyl)-4(3H)-quinazolinone analogues

A novel series of 2-substituted mercapto-3-(3,4,5-trimethoxybenzyl)-4(3H)-quinazolinones 1–20 was synthesised and evaluated for in vitro antitumour activity. N-(4-Chlorophenyl)-2-[(3-(3,4,5-trimethoxybenzyl)-4(3H)-quinazolinon-2-yl)thio)acetamide (7) and N-(3,4,5 trimethoxybenzyl)-2-[(3-(3,4,5-trimethoxybenzyl)-4(3H)-quinazolinon-2-yl)thio]propanamide (19) exhibited excellent antitumour properties, with mean growth inhibitory concentration (GI₅₀) of 17.90 and 6.33 µΜ, respectively, compared with those of 5-fluorouracil 5-FU, gefitinib, and erlotinib (mean GI₅₀: 18.60, 3.24, and 7.29 µΜ, respectively). Comparison of the GI₅₀ (µM) values of compounds 7 and 19 versus those of 5-FU, gefitinib, and erlotinib against an in vitro subpanel of tumour cells lines showed that compounds 7 and 19 have activities almost equal to or higher than that of those standard drugs, especially against lung, CNS, and breast cancer cells. However, compounds 5, 10, 14, 15, 16, 17, and 20 exhibited effective antitumour activity against the different cell lines tested, with growth inhibition percentage (MGI%) of 19, 24, 19, 17, 16, 15, and 16, respectively. A modelling study was performed for compounds 7 and 19 by docking them into the EGFR kinase enzyme to study their mode of binding with the putative binding site.     

Synthesis and biological evaluation of novel (E)-N'-benzylidene hydrazides as novel c-Met inhibitors through fragment based virtual screening

Abstract

C-Met plays a crucial role in the development and progression of neoplastic disease. Type II c-Met inhibitors recognise the inactive DFG-out conformation of the kinase, result in better anti-tumour effects due to synergistic effect against the other kinases. According to our previous works, an (E)-N'-benzylidene group was selected as the initial fragment. Two series of (E)-N'-benzylidene hydrazides were designed by fragment growth method. The inhibitory activities were in vitro investigated against c-Met and VEGFR-2. Compound 10b exhibited the most potent inhibitory activity against the c-Met inhibitor (IC₅₀ = 0.37?nM). Compound 11b exhibited multi-target c-Met kinase inhibitory activity as a potential type II c-Met inhibitor (IC₅₀ = 3.41?nM against c-Met; 25.34?nM against VEGFR-2). The two compounds also demonstrate the feasibility of fragment-based virtual screening method for drug discovery.     

Insecticidal activity, mammalian cytotoxicity and mutagenicity of an ethanolic extract from Nerium oleander (Apocynaceae)

A bioassay procedures utilising the western‐banded blow fly, Chrysomya albiceps (Diptera‐Calliphoridae) has been used to guide the fractionation of an ethanolic extract from the leaves of Nerium oleander (Apocynaceae). The cardiotonic glycoside, neriifolin: (3‐[(6‐deoxy‐3‐0‐methyl‐α‐L glucopyranosyl) oxy]‐14‐hydroxy‐5 β Card‐20 [22]‐endolide) was crystallised from the insecticidal active fraction of the ethanolic extract. The values of LC₅₀ of the ethanolic extract, active fraction, isolated crystals and authentic neriifolin were 164 ppm, 57 ppm, 35 ppm and 36 ppm, respectively when incorporated into the blow fly diet. A primary test on the cytotoxicity and mutagenicity of the crude extract against non‐target organisms was achieved by determining the cytotoxicity and mutagenicity on a mammalian cell line using different concentrations of the extract (in vitro) through a radioactive thymidine incorporation technique and hypoxanthine phosphoribosyl transferase (HPRT) test, using the Chinese hamster lung fibroblasts (V79‐MZ cell line). Cytotoxicity tests revealed that the LC₅₀ was approximately 200 ppm, and the mutagenicity was very low compared to the standard active mutagen.     

Antifeedant and toxic activity of Trichilia americana extract against the larvae of Spodoptera litura

The biological activity of a crude methanolic extract of Trichilia americana (Sesse and Mocino) Pennington (Meliaceae) was assessed using the Asian armyworm, Spodoptera litura (Fabr.) (Lepidoptera: Noctuidae). The extract exhibited strong antifeedant activity in a choice leaf disc bioassay with 0.18 g cm^–2 extract deterring feeding by 50%. In nutritional assays, the crude extract reduced growth, consumption and the utilisation of ingested and digested food in a dose-dependent manner when fed to larvae, suggesting both antifeedant and toxic activities. When relative growth rates were plotted against relative consumption rates, the growth efficiency of the S. litura fed on diet containing T. americana crude extract was significantly less than that of control larvae. This result further indicates that the extract acts as both an antifeedant and chronic toxin. Toxicity is only seen following ingestion and was not observed following topical application or injection into the hemocoel. Larvae reared initially on extract-containing diet then transferred to control diet showed nutritional indices comparable to those of larvae fed continuously on control diet. This suggests that the extract is not permanently damaging the insect's digestive tract. The mode-of-action of the extract as a chronic toxin remains unknown.     

Phagodeterrence and insecticidal activity of Hyptis suaveolens (Poit.) against four important lepidopteran pests

This study was carried out to evaluate the antifeedant and larvicidal activities of Hyptis suaveolens Poit. (Lamiaceae) leaves crude extracts and their fractions against four lepidopteran pests namely Helicoverpa armigera (Hbn.), Spodoptera litura (Fab.), Earias vittella (Fab.) and Leucinodes orbonalis (G.). Hexane, chloroform and ethyl acetate extracts were tested by leaf disc and fruit disc no-choice methods at 1% and 1,000?ppm concentrations for crude and fractions, respectively. Ethyl acetate extract of H. suaveolens exhibited the maximum antifeedant and insecticidal activity at 1% concentration against all the tested insects. It was subjected to fractionation using silica column chromatography with different combinations of hexane and ethyl acetate used as mobile phase. Among the 15 fractions obtained, fraction 2 showed the maximum antifeedant and insecticidal activity against all tested insects at 1000?ppm concentration. Preliminary phytochemical analysis of fraction 2 showed the presence of terpenoids and alkaloids. H. suaveolens could be considered as a safe and eco-friendly insecticide for lepidopteron pest management.     

Larvicidal activity of Pseudocalymma alliaceum and Allium sativum against Culex quinquefasciatus (Say)

The larvicidal activity of the plant extracts Pseudocalymma alliaceum and Allium sativum were determined against Culex quinquefasciatus. The hexane extract of P. alliaceum and the petroleum ether extract of A. sativum exhibited larvicidal efficacy against Cx. quinquefasciatus larvae. Extracts of P. alliaceum resulted in concentrations that produced 50% mortality LC₅₀ and LC₉₀ values of 2.49 and 15.06 ppm, respectively, after 24 h and 1.16 and 8.45 ppm after 48 h. Extracts of A. sativum resulted in LC₅₀ and LC₉₀ values of 8.38 and 29.15 ppm after 24 h and 7.28 and 44.19 ppm after 48 h of exposure, respectively. The results indicate that the plant extract component(s) present in the hexane extract of P. alliaceum leaves demonstrated greater potential as an efficient larvicide than A. sativum against Cx. quinquefasciatus.     

Synthesis and potential antitumor activity of 7-(4-substituted piperazin-1-yl)-4-oxoquinolines based on ciprofloxacin and norfloxacin scaffolds: in silico studies

The potential antitumor activities of a series of 7-(4-substituted piperazin-1-yl)fluoroquinolone derivatives (1–14a,b) using ciprofloxacin and norfloxacin as scaffolds are described. These compounds exhibit potent and broad spectrum antitumor activities using 60 human cell lines in addition to the inherent antibacterial activity. Compounds 1a, 2a, 3b, 6b and 7a were found to be the most potent, while 2b, 5b, and 6a were found to have an average activity. The results of this study demonstrated that compounds 1a, 2a, 3b, 6b and 7a (mean GI₅₀; 2.63–3.09?µM) are nearly 7-fold more potent compared with the positive control 5-fluorouracil (mean GI₅₀; 22.60?µM). More interestingly, compounds 1a, 2a, 3b, 6b and 7a have an almost antitumor activity similar to gefitinib (mean GI₅₀; 3.24?µM) and are nearly 2-fold more potent compared to erlotinib (mean GI₅₀; 7.29?µM). In silico study and ADME-Tox prediction methodology were used to study the antitumor activity of the most active compounds and to identify the structural features required for antitumor activity.     

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.     

Insecticidal and antifungal potentials of a proteinaceous extract of Cassia occidentalis (Linn.) seeds

Proteinaceous extract obtained from Cassia occidentalis seeds with purification fold of 3.91 and 82.7% of bovine trypsin inhibitory activity was assessed at different concentrations (25, 50, 100, 200, 400 and 800 μg/ml) against Spodoptera litura. Assay of larval feeding suggested proteinaceous extract to be toxic as prepupal (80.16%) and pupal mortalities (100%) along with growth deterrent effect with only 16.71% pupation was observed at 800 μg/ml. Fifty per cent mortality (LC₅₀) was observed at 132.91 μg/ml. Also the inhibitor affected fecundity, longevity and percentage of egg hatching. Nutritional indices were adversely affected as both efficiency of conversion of ingested and digested food decreased while approximate digestibility and metabolic cost increased. In vitro studies on proteolytic enzymes of S. litura revealed inhibition of trypsin and chymotrypsin in lumen and faecal matter at all tested concentrations. Also proteinaceous extract inhibited mycelial growth in Fusarium oxysporum, Alternaria brassicicola and Alternaria alternata at 100 μg/ml.     

Growth regulatory and growth inhibitory effects of Thevetia neriifolia stem extracts on Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract

Helicoverpa armigera, a serious global destructive pest of agricultural crops, is on continuous rise despite several control measures undertaken. The detrimental effects of these measures have created a dire need to explore alternate eco-safe strategies. The present study investigates the growth-regulatory and growth-arrest potential of hexane and methanol extracts of Thevetia neriifolia stems against H. armigera. Investigations revealed that larval feeding and rearing on the extract-containing diet did not result in appreciable larval mortality but delayed the larval growth and development. Both the extracts demonstrated dose-dependent effects exhibiting negative correlation between the weights gained by developmental stages and the extract concentrations. Feeding with extracts also resulted in formation of few larval–pupal and pupal–adult intermediates and significantly reduced percent adult emergence of H. armigera. Current study, however, revealed higher growth inhibitory potential of methanol extracts as compared with hexane extracts. The study attempts to provide an eco-friendly approach for H. armigera management.     

Antioxidative iridoid glycosides from the sky flower (Duranta repens Linn)

Phytochemical investigations were performed on the EtOAc-soluble fraction of the whole plant of the sky flower (Duranta repens) which led to the isolation of the iridoid glycosides 1–6. Their structures were elucidated by both 1D and 2D NMR spectroscopic analysis. All the compounds showed potent antioxidative scavenging activity in four different tests, with half maximal inhibitory concentration (IC₅₀) values in the range 0.481–0.719?mM against DPPH radicals, 4.07–17.21 µM for the hydroxyl radical (^?OH) inhibitory activity test, 43.3–97.37 µM in the total reactive oxygen species (ROS) inhibitory activity test, and 3.39–18.94 µM in the peroxynitrite (ONOO^?) scavenging activity test. Duranterectoside A (1) displayed the strongest scavenging potential with IC₅₀ values of (0.481?±?0.06?mM, 4.07?±?0.03, 43.30?±?0.05, 3.39?±?0.02?µM) for the DPPH radicals, ^?OH inhibitory activity test, total ROS inhibitory activity test and the ONOO^? scavenging activity test, respectively.     

Synthesis,antitumor activity and molecular docking study of some novel 3-benzyl-4(3H)quinazolinone analogues

A novel series of 3-benzyl-substituted-4(3H)-quinazolinones were designed, synthesized and evaluated for their in vitro antitumor activity. The results of this study demonstrated that 2-(3-benzyl-6-methyl-4-oxo-3,4-dihydroquinazolin-2-ylthio)-N-(3,4,5-trimethoxyphenyl)acetamide, 2-(3-benzyl-6,7-dimethoxy-4-oxo-3,4-dihydroquinazolin-2-ylthio)-N-(3,4,5-trimethoxyphenyl)acetamide and 3-(3-benzyl-6-methyl-4-oxo-3,4-dihydroquinazolin-2-ylthio)-N-(3,4,5-trimethoxyphenyl)-propanamide have shown amazing broad spectrum antitumor activity with mean GI₅₀ (10.47, 7.24 and 14.12?µM. respectively), and are nearly 1.5–3.0-fold more potent compared with the positive control 5-FU with mean GI₅₀, 22.60?µM. On the other hand, compounds 6 and 10 yielded selective activities toward CNS, renal and breast cancer cell lines, whereas compound 9 showed selective activities towards leukemia cell lines. Molecular docking methodology was performed for compounds 7 and 8 into ATP binding site of EGFR-TK which showed similar binding mode to erlotinib, while compound 11 into ATP binding site of B-RAF kinase inhibited the growth of melanoma cell lines through inhibition of B-RAF kinase, similar to PLX4032.     

Antifeedant and larvicidal activities of Rhein isolated from the flowers of Cassia fistula L.

Antifeedant and larvicidal activities of rhein (1,8-dihydroxyanthraquinone-3-carboxylic acid) isolated from the ethyl acetate extract of Cassia fistula flower were studied against lepidopteron pests Spodoptera litura and Helicoverpa armigera. Significant antifeedant activity was observed against H. armigera (76.13%) at 1000 ppm concentration. Rhein exhibited larvicidal activity against H. armigera (67.5), S. litura (36.25%) and the LC₅₀ values was 606.50 ppm for H. armigera and 1192.55 ppm for S.litura. The survived larvae produced malformed adults.     

Enhanced bioefficacy of Bacillus thuringiensis var. kurstaki against Spodoptera litura (Lepidoptera: Noctuidae) through particle size reduction and formulation as a suspension concentrate

Technical powder of DOR Bt-127, a local isolate of Bacillus thuringiensis (Bt) var. kurstaki, containing 105?µm particles was subjected to milling in a planetary ball mill. Dynamic light scattering of samples drawn at 30?min intervals revealed reduced particle sizes ranging 1171–210?nm. Alkali-soluble protein was higher at 153–175?mg/g in milled Bt powders over 140?mg/g in unmilled Bt powder. All milled powders gave higher mortality of Spodoptera litura (Fabricius) larvae in comparison to the unmilled Bt powder. Bt powder from 60?min milling containing 559?nm particles with a low polydispersity index of 0.338 gave a lower LC₅₀ value of 1.35?mg/ml against third instar S. litura larvae in comparison to 2.04?mg/ml for unmilled Bt powder. LC₅₀ value of the suspension concentrate (SC) formulation developed with 559?nm Bt particles was 2.84?µl/ml containing only 0.95?mg Bt. Field evaluation of the SC formulation against S. litura on castor revealed highest per cent larval reduction of 92.4 and 96.2 at concentrations of 2.5 and 3.0?ml/l, respectively.