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.     

Phytochemical analysis and fabrication of silver nanoparticles using Acacia catechu: An efficacious and ecofriendly control tool against selected polyphagous insect pests

Globally, the farmers are struggling with polyphagous insect pest, and it is the number one enemy of agri-products, which made plenty of economic deterioration. Spodoptera litura and Helicoverpa armigera are the agronomically important polyphagous pests. Most of the farmers are predominately dependent on synthetic chemical insecticides (SCIs) for battle against polyphagous pets. As a result, the broad spectrum usage of SCIs led a lot of detrimental outcomes only inconsequently the researchers search the former-friendly phyto-pesticidal approach. In the present investigation, leaf ethanol extract (LEE) and silver nanoparticles (AgNPs) of A. catechu (Ac) were subjected to various spectral (TLC, CC, UV, FTIR, XRD and SEM) analyses. Larval and pupal toxicity of A. catechu Ac-LEE and Ac-AgNPs were tested against selected polyphagous insect pests. The significant larval and pupal toxicity were experimentally proven, and the highest toxicity noticed in AgNPs than Ac-LEE. The larval and pupal toxicity of Ac-AgNPs tested against S. litura and H. armigera LC₅₀/LC₉₀ values were 71.04/ 74.78, 85.33/ 88.91 µg/mL and 92.57/ 96.21 and 124.43/ 129.95 µg/mL respectively. Ac-AgNPs could be potential phyto-pesticidal effectiveness against selected polyphagous insect pests. In globally, it is significantly sufficient ratification giving towards the prevention of many unauthorized SCPs.     

Biological Effects of Caesalpinia crista Seed Extracts on Helicoverpa armigera (Lepidoptera: Noctuidae) and Its Predator,Coccinella septumpunctata (Coleoptera: Coccinellidae)

Caesalpinia crista seed extracts were investigated in the laboratory against Helicoverpa armigera (Hubner). The extracts exhibited powerful antifeedant and growth disruption activity. The toxic symptoms of the poisoned larvae included reduction in weight gain and mortality of larvae and pupae, larval-pupal intermediates and malformed adults. Maximum anti-feedance is caused by methanol extract (AI₅₀=0.0186%), followed by hexane extract (0.0212%), ethyl acetate extract (0.0416%), butanol extract (0.0767%) and aqueous extract (0.2199%). The larval mortality ranged from 10.00 to 70.00% in different extracts. The 50% larval growth inhibition recorded at 3DAT was statistically at par by methanol and hexane extract. The percent 150 values for inhibiting normal adult emergence were 0.0287, 0.0325, 0.0485, 0.0977 and 0.0547% for methanol, hexane, ethyl acetate, aqueous and butanol extract. The biosafety evaluation of these extracts carried out against the predator, Coccinella septumpunctata showed no mortality of the adults till nine days after treatment. Though the observation taken at 10 DAT, showed slight mortality of adults by methanol extract at both 5.0 and 1.0% concentration.     

Bio-efficacy potential of seaweed Gracilaria firma with copepod,Megacyclops formosanus for the control larvae of dengue vector Aedes aegypti

Mosquitoes are the most critical group of insects in the context of public health, because they transmit numerous diseases, causing millions of deaths annually. The frequent use of systemic insecticides to manage insect pests leads to the destabilization of ecosystems and enhanced resistance to insecticides by pests, suggesting a clear need for alternatives. Marine organisms are a rich source of structurally novel and biologically active metabolites, and cyclopoid copepods are prominent predators in many aquatic ecosystems and have been used as biological agents in successful programs to control mosquito larvae. In this study, we determine the effectiveness of the Taiwanese seaweed Gracilaria firma and different solvent extracts combined with the copepod Megacyclops formosanus for controlling Aedes aegypti. A significant larvicidal potential was recorded after seaweed extract treatment against the dengue vector A. aegypti. Larval mortality was observed after 24 h of exposure in laboratory. All extracts exhibited larvicidal effects; however, the highest larval mortality was observed in the methanol extract of G. firma against A. aegypti larvae (LC₅₀ = 0.251%). The methanol extract of G. firma was more effective than the other extracts and is an ideal eco-friendly approach for the control larvae of the dengue vector A. aegypti.     

Joint Insecticidal Action of Bacillus thuringiensis kurstaki with Annona squamosa L. and Prosopis juliflora Swartz DC on Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

The efficacy of methanolic seed extracts of Annona squamosa (As) and Prosopis juliflora (Pj) individually and in combination with spore-crystal mixture of Bacillus thuringiensis kurstaki (Btk) was tested against third instar larvae of Helicoverpa armigera. The combination of botanicals with Btk significantly decreased LC₅₀ values in less time as compared to individual treatments. As and Pj in combination with Btk significantly affected feeding and dietary utilization of H. armigera significantly than in individual treatments. Physiological effects found in larvae fed on Btk and botanicals included extended larval and pupal durations, decreased adult longevity and fecundity, difficulty in moulting between various instars, formation of larval–pupal intermediates and abnormal pupae. As was found to have a higher interaction with Btk than Pj. Further, Btk showed strong synergistic action with As and Pj, which are the promising candidates for development of alternative bioinsecticide in IPM programs.     

The toxicity and biological effect of Pedalium murex L. extracts on the tobacco cutworm,Spodoptera litura (Fabr.) larvae

Petroleum ether, acetone, water and aqueous extract of Pedalium murex L. (Pedalaceae) leaves, root and fruit were tested against fourth instar larvae of the tobacco cutworm, Spodoptera litura (Fabr.) under laboratory conditions using leaf dip method. Larval mortality; larval, pupal and adult periods; pupal weight, pupation and adult emergence; larval and pupal deformities of S. litura were recorded. All the tested solvent extracts of P. murex were effective against S. litura life stages by causing mortality in a dose dependent manner. However, the efficacy was more significant with respect to acetone (leaf and root) and petroleum ether (root) at higher concentrations (0.8%) which leads to 100% larval mortality. The water and aqueous extracts of root caused 86.6 and 88.0% larval mortality respectively at 4% concentration. Among the plant parts tested, root had more importance followed by leaf and fruit. Plant extracts extended the larval, pupal and adult periods; reduced the pupal weight, pupal and adult emergence and caused larval–pupal deformities. Total life time was highly prolonged in acetone extracts of P. murex root. Though all the extracts were found to have insecticide activity, acetone and petroleum ether extracts of P. murex root can be used as an effective alternative to modern synthetic insecticides. Bioactive principles from these extracts can be isolated, identified and integrated in S. litura management.     

Chemical compositions of leaf extracts from Conocarpus erectus L. (Combretaceae) and their bioactivities against Tribolium castaneum Herbst (Coleoptera: Tenebrionidae)

Botanical insecticides have long been considered as alternatives to synthetic chemical insecticides in IPM programs. Effects of aqueous and ethanolic extracts obtained from buttonwood, Conocarpus erectus L. (Combretaceae), leaves on a major kind of stored product pests, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae), were evaluated under laboratory conditions. For this purpose, LC₅₀, repellency, antifeedant properties, and some biological effects (including body weight, immature developmental time and survival) of the insect were determined. The aqueous and ethanolic extracts were highly toxic to the larvae and adults. Calculated LC₅₀ values ranged between 2.6 and 193.4 (g/kg). Both extracts had repellent and antifeedant properties against the adults. The extracts adversely affected the larval and pupal weights, developmental time, and survival. Aqueous extracts were more effective only for the LC₅₀ values and only in females. All other measured parameters do not differ between the two extracts. The bioactive properties might be related to high amounts of alkaloid, phenol and tannin. Aqueous extract of the plant leaves may be a useful alternative for chemical insecticides.     

The life parameters of a parasitoid Microplitis mediator (Hymenoptera: Braconidae), reared on cotton bollworm Helicoverpa armigera (Hübner) with Cry1Ac diet

Microplitis mediator (Haliday) is an important endoparasitoid of the cotton bollworm, Helicoverpa armigera (Hübner) in northern China. Interactions among H. armigera, its larval parasitoid M. mediator, and Cry1Ac over two generations were evaluated under laboratory conditions. The results indicated that the developmental period of M. mediator offspring's eggs and larvae were significantly delayed and pupal and adult weight were significantly less compared to the control when the female parasitoids parasitized H. armigera larvae that fed on diet containing 1, 2, 4 and 8 µg/g Cry1Ac. The female parasitoids emerged from the host fed diet containing 8 µg/g Cry1Ac could oviposit in healthy hosts, and their offspring's biological parameters (egg–larval period, pupal weight and adult weight), parasitism rate, abnormal cocoon rate and adult emergence were not significantly affected. Cry1Ac was detected in larvae and hemolymph of H. armigera, but not in the larvae of M. mediator. The results suggest that the observed significant effects on several fitness parameters of the F₁ M. mediator developed from H. armigera fed Cry1Ac diet most likely were host-quality mediated rather than direct effects of Cry1Ac.     

Biopotential of Fleuggea leucopyrus (Koen.) Willd. against Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

In the past five decades, agricultural pests are controlled by synthetic pesticides which caused ill effects on non-target organisms and environment; also insect pests developed resistance and minor pests became major pests. By keeping this in mind, the present study was selected. Antifeedant and larvicidal activities and growth inhibitory effects of hexane, chloroform and ethyl acetate leaf extracts of Fleuggea leucopyrus (Koen.) Willd. against Helicoverpa armigera (Hbn.) were studied. Antifeedant and larvicidal activities were evaluated at 0.5, 1, 2.5 and 5% concentrations at laboratory conditions. Hexane and chloroform extracts at 5% concentration recorded significant antifeedant activity of 82.41 and 74.45%, respectively. Hexane extract recorded the least LC₅₀ and LC₉₀ values of 1.37 and 4.80%, respectively. Hexane extract recorded maximum larvicidal activity of 85.78% with LC₅₀ and LC₉₀ values of 1.69 and 4.94%, respectively. Ethyl acetate extract also had notable amount of larvicidal activity of 81.77% at 5% concentration. Hexane extract at 5 and 2.5% and ethyl acetate extract at 5% concentrations completely prevented the adult emergence of H. armigera. This plant could be further exploited to identify the active principle(s) responsible for the activities and to develop a novel pesticidal formulation. This is the first report of F. leucopyrus is studied for its bioefficacy against H. armigera.     

Pupal diapause of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) mediated by larval host plants: pupal weight is important

Facultative diapause, a strategy that allows insects to initiate additional generations when conditions are favorable or to enter diapause when they are not, has a profound effect on the ecology and evolution of species. Most previous studies have concentrated on the role of photoperiod and temperature in inducing facultative diapause in insects. In contrast, here we studied pupal diapause mediated by larval host plants in the cotton bollworm Helicoverpa armigera, and confirmed that pupal weight is a critical factor. Two groups of third instar H. armigera larvae, kept at 25 °C with L:D = 8:16 and 20 °C with photoperiod of L:D = 8:16, respectively, were fed on six host plants and on artificial diet (as a control) to determine how larval host plants affect diapause incidence and related traits (such as pupal weight and developmental duration). The data showed larval host plants affected diapause incidence significantly and the effects could be masked by low temperature. Further analysis showed that pupal size, not the length of the sensitive stage, affected the decision to enter diapause. In a further experiment, third-instar to final-stage larvae deprived of artificial diet for 2 days demonstrated a direct relationship between pupal weight and diapause incidence. These results suggest that larval host plants, by affecting pupal size, may influence diapause occurrence in H. armigera. This has important adaptive significance for both over-wintering survival and the possibility for completing an additional generation.     

Manifold passages in an assorted infection in a host could improve virulence of Helicoverpa armigera Nucleopolyhedrovirus (HaNPV)

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is serious pests of cotton and several other crops. Helicoverpa armigera Nucleopolyhedrovirus (HaNPV) can be important alternative to synthetic insecticides for the management of H. armigera. However, the efficacy of HaNPV can vary in horizontal and vertical transmission. In the current study, we evaluated the efficacy of HaNPV of a virulent strain (vertically transmitted up to six generations) and wild strains (used after isolation from the field infected larvae). Both strains were applied to the 2nd instar larvae of H. armigera @ 1 × 10⁹ polyhedral inclusion bodies (PIB)/ml. There were six replications of each strain (strains). The results indicated higher mortalities in larvae exposed to virulent strains (68.33 ± 6.07%) as compared to wild strain (45 ± 2.24%). Virulent strains killed the larvae quite faster than wild strain. The lethal time (LT₅₀) to kill 50% of the larvae by virulent strain was 7.15 days and for wild strain it was 19.47 days. The results showed that multiple passage of HaNPV through several generations enhances its efficacy to kill H. armigera larvae faster. The results of this study will be helpful to manage H. armigera and other related lepidopoterous pests.     

A simple and reliable method for discriminating between Helicoverpa armigera and Helicoverpa assulta (Lepidoptera: Noctuidae)

Abstract The cotton bollworm Helicoverpa armigera and the oriental tobacco budworm H. assulta are sibling species, both being important agricultural pests. Morphologically, the two insects are almost indistinguishable at the egg, larval and pupal stages. One of the big challenges in the study of these insects, in particular in integrated pest management, is a timely and dependable identification of these insects at their early stages of development. Here, we report a H. armigera‐specific nuclear DNA marker, and demonstrate that it can be employed to reliably discriminate between H. armigera and H. assulta by simple polymerase chain reaction amplification experiment.     

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.     

Biocontrol Potential of Steinernema thermophilum and Its Symbiont Xenorhabdus indica Against Lepidopteran Pests: Virulence to Egg and Larval Stages

Under laboratory conditions, the biocontrol potential of Steinernema thermophilum was tested against eggs and larval stages of two important lepidopteran insect pests, Helicoverpa armigera and Spodoptera litura (polyphagous pests), as well as Galleria mellonella (used as a model host). In terms of host susceptibility of lepidopteran larvae to S. thermophilum, based on the LC₅₀ 36 hr after treatment, G. mellonella (LC₅₀ = 16.28 IJ/larva) was found to be more susceptible than S. litura (LC₅₀ = 85 IJ/larva), whereas neither host was found to be significantly different from H. armigera (LC₅₀ = 54.68 IJ/larva). In addition to virulence to the larval stages, ovicidal activity up to 84% was observed at 200 IJ/50 and 100 eggs of H. armigera and S. litura, respectively. To our knowledge this is the first report of entomopathogenic nematode pathogenicity to lepidopteran eggs. Production of infective juvenile (IJ) nematodes/insect larva was also measured and found to be positively correlated with rate of IJ for H. armigera (r = 0.990), S. litura (r = 0.892), as well as G. mellonella (r = 0.834). Both Phase I and Phase II of symbiotic bacteria Xenorhabdus indica were tested separately against neonates of H. armigera and S. litura by feeding assays and found to be virulent to the target pests; phase variation did not affect the level of virulence. Thus S. thermophilum as well as the nematode’s symbiotic bacteria applied separately have the potential to be developed as biocontrol agents for key lepidopteran pests.     

Mitochondrial P-Glycoprotein ATPase Contributes to Insecticide Resistance in the Cotton Bollworm,Helicoverpa armigera (Noctuidae: Lepidoptera)

Cotton bollworm, Helicoverpa armigera, is one of the most damaging polyphagous pests worldwide, which has developed high levels of resistance to commonly applied insecticides. Mitochondrial P-glycoprotein (Pgp) was detected in the insecticide-resistant strain of H. armigera using C219 antibodies, and its possible role was demonstrated in the efflux of xenobiotic compounds using spectrofluorometer. The TMR accumulated in mitochondria in the absence of ATP, and effluxed out in presence of ATP; the process of efflux was inhibited in the presence of ortho-vandate, an inhibitor of Pgp, in insecticide-resistant larvae of H. armigera. The mitochondria isolated from insecticide-resistant larvae were resistant to insecticide-induced inhibition of oxygen consumption and cytochrome c release. Membrane potential decreased in a dose-dependent manner in the presence of higher concentration of insecticides (>50 µM) in mitochondria of insecticide-resistant larvae. In conclusion, mitochondrial Pgp ATPase detected in the insecticide-resistant larvae influenced the efflux of xenobiotic compounds. Pgp might be involved in protecting the mitochondrial DNA and the components of the electron transport chain from damage due to insecticides, and contributing to the resistance to the deleterious effects of insecticides on the growth of insecticide-resistant H. armigera larvae.     

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.     

Monitoring of four major lepidopteran pests in Korean cornfields and management of Helicoverpa armigera

Lepidopterans such as Helicoverpa armigera are emerging pests of corn in Korea, causing huge yield losses and deteriorating the quality of the corn crop. We monitored four major lepidopteran pests in major corn growing areas of Korea by employing sex pheromone traps from 2012 to 2015 to understand population dynamics. H. armigera, Ostrinia furnacalis, Spodoptera litura, Spodoptera exigua and Mythimna separata were collected from April to October. Among these species, H. armigera was found to be the main pest based on crop damage intensity (80–90% of the total damage). H. armigera was generally observed during the earing stage of corn and would migrate to other fields during September to October for overwintering. We also tested select insecticides against larvae of H. armigera in the laboratory. The most effective insecticide was indoxacarb, which was applied at specific times to manage H. armigera in cornfields. Optimal timing of spraying was estimated at about 70 days after planting (earing season) to control for H. armigera. Additionally, two species of parasitoid (Therion circumflexum and Ophioninae sp.) were identified from H. armigera. However, more extensive surveys are needed to organize a control program based on natural enemies.     

Morphometric Variation in Geographic Populations of Cotton Bollworm,Helicoverpa armigera (Hübner) Occurring in South Indian Cotton Ecosystems

Morphometric characterization of twelve geographic populations of cotton bollworm, Helicoverpa armigera occurring in south Indian cotton ecosystems was done at larval, pupal and adult stages over three cropping seasons. Traits such as length and weight of larvae, pupa and length and width of the wing, length of fore-, mid- and hind femur, male reproductive organ-length of genital capsule, valves, and ejaculatory duct, female reproductive organ-length of appendix bursae and ductus bursae at adult stage were measured across three years. Populations significantly differed for most of the traits studied. It was evident that populations from northern parts recorded higher phenotypic attributes compared to those from southern parts of south Indian cotton ecosystem. Besides larval, pupal and adult external phenotypic traits, attributes of male reproductive organ viz., length of genital capsule, valves, and ejaculatory duct and female reproductive organ viz., length of appendix bursae and ductus bursae differed significantly among populations. Information on population structure and differences based on morphometry would be useful in better understanding of population dynamics and management of this pest in cotton and other crops in south India.     

A kidney bean trypsin inhibitor with an insecticidal potential against Helicoverpa armigera and Spodoptera litura

In the present study, trypsin inhibitor extracts of ten kidney bean seed (Phaseolus vulgaris) varieties exhibiting trypsin and gut trypsin-like protease inhibitor activity were tested on Helicoverpa armigera and Spodoptera litura. Trypsin inhibitor protein was isolated and purified using multi-step strategy with a recovery of ~15 % and purification fold by ~39.4. SDS-PAGE revealed a single band corresponding to molecular mass of ~15 kDa and inhibitory activity was confirmed by reverse zymogram analyses. The inhibitor retained its inhibitory activity over a broad range of pH (3–11), temperature (40–60 °C) and thermostability was promoted by casein, CaCl₂, BSA and sucrose. The purified inhibitor inhibited bovine trypsin in 1:1 molar ratio. Kinetic studies showed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 1.85 μM. The purified trypsin inhibitor protein was further incorporated in the artificial diet and fed to second instar larvae. A maximum of 91.7 % inhibition was obtained in H. armigera, while it was moderate in S. litura (29 %) with slight varietal differences. The insect bioassay showed 40 and 22 % decrease in larval growth followed by 3 and 2 days delay in pupation of H. armigera and S. litura, respectively. Some of the adults emerged were deformed and not fully formed. Trypsin inhibitor protein was more effective against H. armigera as it showed 46.7 % mortality during larval growth period compared to S. litura (13.3 %).