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Paranosema locustae is an environmentally friendly parasitic predator with promising applications in locust control. In this study, transcriptome sequencing was conducted on gonadal tissues of Locusta migratoria males and females infected and uninfected with P. locustae at different developmental stages. A total of 18,635 differentially expressed genes (DEGs) were identified in female ovary tissue transcriptomes, with the highest number of DEGs observed at 1 day post-eclosion (7141). In male testis tissue transcriptomes, a total of 32,954 DEGs were identified, with the highest number observed at 9 days post-eclosion (11,245). Venn analysis revealed 25 common DEGs among female groups and 205 common DEGs among male groups. Gene ontology and Kyoto Encyclopaedia of Genes and Genome analyses indicated that DEGs were mainly enriched in basic metabolism such as amino acid metabolism, carbohydrate metabolism, lipid metabolism, and immune response processes. Protein-protein interaction analysis results indicated that L. migratoria regulates the expression of immune- and reproductive-related genes to meet the body's demands in different developmental stages after P. locustae infection. Immune- and reproductive-related genes in L. migratoria gonadal tissue were screened based on database annotation information and relevant literature. Genes such as Tsf , Hex1 , Apolp-III , Serpin , Defense , Hsp70 , Hsp90 , JHBP , JHE , JHEH1 , JHAMT , and VgR play important roles in the balance between immune response and reproduction in gonadal tissues. For transcriptome validation, Tsf , Hex1 , and ApoLp-III were selected and verified by quantitative real-time polymerase chain reaction (qRT-PCR). Correlation analysis revealed that the qRT-PCR expression patterns were consistent with the RNA-Seq results. These findings contribute to further understanding the interaction mechanisms between locusts and P. locustae .

In natural environments, general plant volatiles and herbivore-induced plant volatiles (HIPVs) serve as critical clues for predatory natural enemies in the search for prey. The insect olfactory system plays a vital role in perceiving plant volatiles including HIPVs. In this study, we found that HIPV ( E , E )-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT) and the plant volatile geranyl acetate (GA), two structurally similar chemicals, displayed electrophysiological activities on the antennae of the ladybird Hippodamia variegata , but were only attractive to adult females in behavior. Moreover, mated female ladybirds laid a significantly higher number of eggs on TMTT-treated and GA-treated cotton leaves compared to controls. Screening of female-biased odorant receptors (ORs) from the antennal transcriptomes, performing Xenopus oocytes expression coupled with two-electrode voltage clamp recordings, suggested that HvarOR28 specifically tuned to TMTT and GA. Molecular docking and site-directed mutagenesis revealed that the amino acid residues Tyr143 and Phe81 of HvarOR28 are the key site for binding with TMTT and GA. Furthermore, RNA interference (RNAi) assay demonstrated that HvarOR28 -silenced individuals demonstrated a notable decrease in electrophysiological responses, even female adults almost lost behavioral preference for the two compounds. Thus, it could be concluded that HvarOR28 in H. variegata contributes to facilitating egg laying through the perception of TMTT and GA. These findings may help to develop new olfactory modulators based on the behaviorally active ligands of HvarOR28.

Botanical pesticides are one of the most promising alternatives to synthetic insecticides for green pest management. However, their efficacies must be further improved to meet real needs. Here we designed a nanoscale camptothecin (CPT) encapsulated in a star polycation (SPc) and determined its bioactivity against a devastating agricultural pest, Spodoptera frugiperda. The self-assembly of CPT/SPc complex was mainly driven by hydrogen bonding and Van der Waals forces to decrease the particle size from 789 to 298 nm. With the help of SPc, the contact angle of CPT decreased from 116° to 92° on maize leaves, and its retention was increased from 5.53 to 11.97 mg/cm 2 . The stability of SPc-loaded CPT was also improved in an alkaline environment, which is beneficial for its acting in lepidopteran insect guts. The CPT/SPc complex had stronger larvicidal activity and ovicidal activity against S. frugiperda than CPT alone, led to more complex transcriptomic changes in larvae, and had obvious adverse impacts on the activities of two digestive enzymes. Our findings demonstrated that the encapsulation of CPT by SPc-based nanodelivery system enabled better insecticidal activities against S. frugiperda, which holds great promise for the development of more efficient and sustainable pest control strategies to meet the demands of modern crop protection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Entomophagous fungi (EPF) not only directly kill insect pests, but also colonize plants and improve their resistance against pests. However, most previous research has focused on Beauveria bassiana and Metarhizium anisopliae, and there are few reports on whether other EPF can enhance resistance against pests via endogenous colonization. Herein, an EPF strain was isolated from diseased larvae of Spodoptera litura in a soybean field, and subjected to genome-wide sequencing at the chromosomal level. The pathogenicity of the isolate toward various pest insects was evaluated, and the ability to colonize plants and induce resistance against phytopathogens and insect pests was tested., Results: The purified isolate was identified as M. rileyi and designated MrS1Gz1-1. Biological assays revealed its strong pathogenicity toward five insect pests belonging to Lepidoptera and Hemiptera. Furthermore, the strain inhibited the growth of soil-borne plant disease caused by Sclerotinia sclerotiorum in vitro. It colonized plants as an endophyte via soil application, thereby inducing plant resistance-related genes against phytopathogen infection, and it disrupted the feeding selectivity of S. litura larvae., Conclusion: M. rileyi MrS1Gz1-1 has potential as a broad-spectrum microbial control agent that can induce resistance against phytopathogens and insect pests feeding as an endotype. The complete genome provides a valuable resource for exploring host interactions. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Megalurothrips usitatus (Bagnall) (Thysanoptera: Thripidae) is an important pest in Vigna unguiculata (L.) Walp. Neoseiulus barkeri (Hughes) (Acari: Phytoseiidae) is widely used for control of pest mites and insects worldwide. We evaluated its effect on M. usitatus when predators (N. barkeri) or insecticides (Spinetoram) were applied in the fields. Neoseiulus barkeri Hughes consumed 80% of M. usitatus prey offered within 6 h, and predation showed Type III functional response with prey density. The maximum consumption of N. barkeri was 27.29 ± 1.02 individuals per d per arena (1.5 cm diameter), while the optimal prey density for the predatory mite was 10.35 ± 0.68 individuals per d per arena (1.5 cm diameter). The developmental duration of N. barkeri fed with M. usitatus was significantly shorter than those fed with the dried fruit mite, Carpoglyphus lactis (L.) (Acari: Astigmata). In field trials, the efficiency of N. barkeri against M. usitatus was not significantly different from that of applications of the insecticide spinetoram. Biodiversity of other insects in treated fields was assessed, and there were 21 insect species in garden plots treated with N. barkeri releases. The total abundance (N), Shannon's diversity index (H), Pielou's evenness index (J) and Simpson's diversity index (D) of the garden plots treated with predatory mites were all significantly higher than that in the garden plots treated with spinetoram, where we found no species of predators or parasitoids and 7 herbivores. Our results show that N. barkeri is a potential means to control M. usitatus while preserving arthropod diversity at the level of treated gardens., (© 2024. The Author(s).)

Currently, insecticides that target nicotinic acetylcholine receptors (nAChR) are widely used. Studies on the sublethal effects of insecticides have found that they can affect the amount of virus in insects. The mechanism by which insecticides affect insect virus load remain unclear. Here, we show that nAChR targeting insecticide can affect viral replication through the immune deficiency (IMD) pathway. We demonstrate that a low dose of spinosad (6.8 ng/mL), acting as an antagonist to Drosophila melanogaster nicotinic acetylcholine receptor α6 (Dα6), significantly elevates Drosophila melanogaster sigmavirus (DMelSV) virus titers in adults of Drosophila melanogaster . Conversely, a high dose of spinosad (50 ng/mL), acting as an agonist to Dα6, substantially decreases viral load. This bidirectional regulation of virus levels is absent in Dα6 -knockout flies, signifying the specificity of spinosad's action through Dα6 . Furthermore, the knockdown of Dα6 results in decreased expression of genes in the IMD pathway, including dredd , imd , relish , and downstream antimicrobial peptide genes AttA and AttB , indicating a reduced innate immune response. Subsequent investigations reveal no significant difference in viral titers between relish mutant flies and Dα6 - relish double mutants, suggesting that the IMD pathway's role in antiviral defense is dependent on Dα6 . Collectively, our findings shed light on the intricate interplay between nAChR signaling and the IMD pathway in mediating antiviral immunity, highlighting the potential for nAChR-targeting compounds to inadvertently influence viral dynamics in insect hosts. This knowledge may inform the development of integrated pest management strategies that consider the broader ecological impact of insecticide use.

Introduction: Aspergillus nomiae is known as a pathogenic fungus that infects humans and plants but has never been reported as an entomophagous fungus (EPF) that can provide other functions as an endotype., Methods: A strain of EPF was isolated and identified from diseased larvae of Spodoptera litura in a soybean field and designated AnS1Gzl-1. Pathogenicity of the strain toward various insect pests was evaluated, especially the ability to colonize plants and induce resistance against phytopathogens and insect pests., Results: The isolated EPF strain AnS1Gzl-1 was identified as A. nomiae ; it showed strong pathogenicity toward five insect pests belonging to Lepidoptera and Hemiptera. Furthermore, the strain inhibited the growth of Sclerotinia sclerotiorum in vitro , a causal agent of soil-borne plant disease. It colonized plants as an endophyte via root irrigation with a high colonization rate of 90%, thereby inducing plant resistance against phytopathogen infection, and disrupting the feeding selectivity of S. litura larvae., Discussion: This is the first record of a natural infection of A. nomiae on insects. A. nomiae has the potential to be used as a dual biocontrol EPF because of its ability to not only kill a broad spectrum of insect pests directly but also induce resistance against phytopathogens via plant colonization., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhang, Tian, Sui, Lu, Cheng, Zhao, Li and Shi.)

Microsporidia are a group of eukaryotic intracellular parasitic organisms that infect almost all vertebrates and invertebrates. Paranosema locustae are specialized parasites of Orthoptera that are often used as biological controls of locusts, with slow effects of action. In this study, we found that after infection with P. locustae, changes in energy metabolism in male and female Locusta migratoria as were consistent, with no gender differences. During the first 8 days of infection, L. migratoria used sugar as a source of energy. After 8 days, lipids and proteins were consumed to provide energy when the spore load was considerably heavy, and energy supply was insufficient. With increasing infection concentration and time, energy conversion from sugar, fats, and proteins was improved, which may explain why high mortality did not occur until about 15 days after P. locustae infection. The tandem mass tag-based quantitative proteomics analysis revealed that most altered metabolism-related proteins were upregulated (27 of 29 in the metabolic pathway). This result suggests that P. locustae infection accelerated metabolism in L. migratoria, which facilitated the pathogen's life cycle, inhibiting the growth and development of the locusts and eventually killing them. Our findings will be useful to better understand of the chronic pathogenic mechanisms of P. locustae and inform on applications of P. locustae to control locusts., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America.)

Locusta migratoria is a serious agricultural pest in China. Beauveria bassiana is one of the most important pathogens of grasshoppers and locusts. The effects of ultraviolet light were evaluated on the B. bassiana strain BbZJ1. The results showed that 253.7 and 360 nm wavelength UV (Ultra Violet) did not affect the germination of B. bassiana after its recovery from UV treatments. Nevertheless, the virulence of B. bassiana BbZJ1 after its recovery from radiation of UV (253.7 nm) increased. The mortality rates were 85.00% for the BbZJ1 control, was 96.67% for BbZJ1 recovered from radiation of UV (253.7 nm) for 60 min. After treatment with 253.7 nm UV radiation for 60 min, the expression levels of stress-resistant genes BbAlg9 and Bbadh2 in BbZJ1 strain were 2.68 and 2.29 times higher than those in the control group, respectively. Meanwhile, the B. bassiana prepared in 5% groundnut oil showed highest tolerance levels to the ultraviolet radiation. The 5% groundnut oil was the most suitable potential UV-protectant for B. bassiana in terms of cost and availability., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Polymycoviridae is a recently established family of mycoviruses. Beauveria bassiana polymycovirus 4 (BbPmV-4) was previously reported. However, the effect of the virus on host fungus B. bassiana was not clarified. Here, a comparison between virus-free and virus-infected isogenic lines of B. bassiana revealed that BbPmV-4 infection of B. bassiana changes morphology and could lead to decreases in conidiation and increases in virulence against Ostrinia furnacalis larvae. The differential expression of genes between virus-free and virus-infected strains was compared by RNA-Seq and was consistent with the phenotype of B. bassiana. The enhanced pathogenicity may be related to the significant up-regulation of genes encoding mitogen activated protein kinase, cytochrome P450, and polyketide synthase. The results enable studies of the mechanism of interaction between BbPmV-4 and B. bassiana., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Vigna unguiculata is a vital vegetable crop in Southeast Asia, and Megalurothrips usitatus can cause huge damage to this crop. Enhancing the resistance of V . unguiculata against M . usitatus is a promising way to protect this crop; however, there is limited information regarding the mechanism underlying the resistance of V . unguiculata against M . usitatus . Here, a behavior assay was performed to explore the resistance of V. unguiculata against M. usitatus after insect damage or treatment by jasmonic acid (JA). Furthermore, transcriptome and metabonomics analysis was used to detect the putative mechanism underlying the resistance of V. unguiculata against M. usitatus . The pre-treatment of Vigna unguiculata with JA or infestation with Megalurothrips usitatus alleviated the damage resulting from the pest insect. We further identified differentially expressed genes and different metabolites involved in flavonoid biosynthesis and alpha-linolenic acid metabolism. Genes of chalcone reductase and shikimate O-hydroxycinnamoyltransferase involved in flavonoid biosynthesis, as well as lipoxygenase and acyl-CoA oxidase involved in alpha-linolenic acid metabolism, were upregulated in plants after herbivory or JA supplementation. The upregulation of these genes contributed to the high accumulation of metabolites involved in flavonoid biosynthesis and the alpha-linolenic acid metabolism pathway. These transcriptional and metabolite changes are potentially responsible for plant defense and a putative regulatory model is thus proposed to illustrate the cowpea defense mechanism against insect attack. Our study provides candidate targets for the breeding of varieties with resistance to insect herbivory by molecular technology.

The stability of symbiotic flora is an important indicator of the health of an organism. Symbiotic bacteria have been proven to be closely involved in the immune process of organisms. The pathogenicity of Beauveria bassiana was studied in relation to symbiotic bacteria on the surface and inside of the migratory locust ( Locusta migratoria ). The results showed that the surface disinfection of test locusts contributed to the pathogenicity of B. bassiana to locusts. Most of the surface bacteria of L. migratoria caused some inhibition of B. bassiana growth, and LM5-4 ( Raoultella ornithinolytica ), LM5-2 ( Enterobacter aerogenes ), and LM5-13 ( Citrobacter freundii ) showed the highest inhibitory effect on the growth of B. bassiana. The inoculation of locusts with additional surface symbiotic bacteria reduced the virulence of B. bassiana to L. migratoria . Infection by different strains of B. bassiana caused similar changes in the symbiotic flora of migratory locusts. The inoculation of locusts with additional intestinal symbiotic bacteria ( Enterobacter sp.) reduced the virulence of B. bassiana to L. migratoria . These findings illustrate the effect of bacterial communities on fungal infections in L. migratoria when seen from the perspective of ecology in a microenvironment. The active antifungal substances of such bacteria and their mechanisms of action need further study.

Grasshoppers can swarm in the millions and destroy crops over wide areas, posing a major economic threat to agriculture. A wide range of insect-related viruses has recently been reported in the metagenomics of grasshoppers. Here, we identified and isolated a novel reovirus from grasshoppers, named Acrididae reovirus (ARV). The complete genome of ARV was composed of nine dsRNA segments. Phylogenetic analysis revealed that ARV formed a monophyletic lineage with unclassified insect-associated reoviruses and was sufficiently distinct from known genera of Reoviridae . ARV could replicate in its host Locusta migratoria and result in host death. Lower-dose ARV infection affected ovary development and resulted in a significant reduction in fecundity. The identification and characterization of a novel pathogenic reovirus could potentially promote the development of new biological control agents.

Orius sauteri (Poppius) is an important predator of many economically important insect pests. The mass rearing of O. sauteri is difficult, limiting its application in pest control. Here we assessed the nutritional quality of eggs of Sitotroga cerealella (Olivier), Agrotis ypsilon (Rottemberg), or Spodoptera litura (Fabricius), and their potential for rearing O. sauteri in the laboratory for two generations. Of species tested, S. cerealella eggs resulted in the highest survival and reproduction of O. sauteri compared to the other two lepidopteran species. Eggs of A. ypsilon were a suitable diet for the nymphal stage, which developed faster on A. ysilon eggs than those of S. cerealella eggs. Conversely, eggs of S. litura were not a suitable diet for O. sauteri, and they disrupted the development and reproduction of O. sauteri. Sitotroga cerealella eggs showed advantages in all the nutritional components evaluated. Orius sauteri fed S. litura eggs contained significantly lower protein levels than those fed on the other eggs tested. Spodoptera litura eggs significantly enhanced the CAT activity in O. sauteri, which suggests that some components from S. litura eggs harmed the development and reproduction of O. sauteri. Based on these results, we suggest using a combined diet for mass rearing of the pirate bug, feeding the nymphs and adults with A. ypsilon eggs and S. cerealella eggs, respectively. This study contributes to the discovery of artificial diets for mass rearing O. sauteri and other Orius species in the future., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Mycoviruses can spread interspecifically and intraspecifically in plant pathogenic fungi, as well as spreading intraspecifically in entomogenous fungi, especially Beauveria bassiana. However, whether mycoviruses are common in Beauveria spp. and can spread interspecifically between Beauveria species are unclear. Herein, four Beauveria species, but not B. bassiana, were randomly selected for double stranded RNA (dsRNA) detection. Furthermore, two previously reported dsRNA mycoviruses from B. bassiana, BbCV-2 and BbPmV-4, were used to study the interspecific transmission among B. bassiana, B. amorpha, and B. aranearum, using hyphal anastomosis and a novel insect coinfection transmission method. The results showed that dsRNA mycoviruses exist universally in Beauveria spp. and could spread interspecifically between different Beauveria species. The transmission efficiency from B. bassiana to the other two Beauveria species was significantly higher than that of the reverse transmission. Both viruses could stably and vertically spread in B. amorpha and B. aranearum, which affected their growth rate and colony morphology., Competing Interests: Conflict of interest The authors declare that they have no conflict., (Copyright © 2022. Published by Elsevier B.V.)

( E )-β-ocimene, a ubiquitous monoterpene volatile in plants, is emitted from flowers to attract pollinators and/or from vegetative tissues as part of inducible defenses mediated by complex signaling networks when plants are attacked by insect herbivores. Wild pear species Pyrus betuleafolia used worldwide as rootstock generally displays valuable pest-resistant traits and is a promising genetic resource for pear breeding. In the current study, transcriptional changes in this wild pear species infested with a polyphagous herbivore Spodoptera litura and the underlying molecular mechanisms were fully investigated. A total of 3,118 differentially expressed genes (DEGs) were identified in damaged pear leaf samples. Spodoptera litura larvae infestation activated complex phytohormonal signaling networks in which jasmonic acid, ethylene, brassinosteroids, cytokinin, gibberellic acid and auxin pathways were induced, whereas salicylic acid and abscisic acid pathways were suppressed. All DEGs associated with growth-related photosynthesis were significantly downregulated, whereas most DEGs involved in defense-related early signaling events, transcription factors, green leaf volatiles and volatile terpenes were significantly upregulated. The PbeOCS (GWHGAAYT028729), a putative ( E )-β-ocimene synthase gene, was newly identified in P. betuleafolia transcriptome. The upregulation of PbeOCS in S. litura -infested pear leaves supports a potential role for PbeOCS in herbivore-induced plant defenses. In enzyme-catalyzed reaction, recombinant PbeOCS utilized only geranyl pyrophosphate but not neryl diphosphate, farnesyl pyrophosphate or geranylgeranyl diphosphate as a substrate, producing ( E )-β-ocimene as the major product and a trace amount of ( Z )-β-ocimene. Moreover, as a catalytic product of PbeOCS, ( E )-β-ocimene showed repellent effects on larvae of S. litura in dual-choice bioassays. What is more, ( E )-β-ocimene increased mortalities of larvae in no-choice bioassays. These findings provide an overview of transcriptomic changes in wild pears in response to chewing herbivores and insights into ( E )-β-ocimene biosynthesis in pear plants, which will help elucidate the molecular mechanisms underlying pear-insect interactions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Huang, Zhang, Li, Wang, Guo, Liu, Han, Zhen, Li, Shi and Zhang.)

The widespread cultivation of genetically modified (GM) crops has raised concerns for their safety. Here, we evaluated the effects of a GM maize variety expressing the Cry1Ab (14.76 ± 0.87 μg/g FW) and EPSPS proteins (191.55 ± 15.69 μg/g FW) on the life-history traits and gut bacterial community of a non-target arthropod, Locusta migratoria, in the laboratory. We found that GM maize had no significant effect on the survival or body weight of different development stages of L. migratoria. The midgut and hindgut bacterial diversities and compositions were determined using high-throughput sequencing targeting the V3-V4 regions of the 16S rRNA. No significant changes were found in the species diversity or abundance between insects in the GM-fed treatment and the non-GM control. Furthermore, the concentration of Cry1Ab and EPSPS in the gut was determined after digestion of GM maize. Results showed that the contents of Cry1Ab/EPSPS rapidly decreased and were hard to detect after 72 h. Based on the parameters assessed, we can conclude that the GM maize variety examined has no significant adverse effect on L. migratoria., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Harmonia axyridis Pallas (Coleoptera: Coccinellidae) is an important natural enemy of aphids. Plant species and plant health conditions can affect the behavior of H. axyridis. To determine plant effects on this lady beetle, we examined beetle responses to four cover crops: coriander (Coriadrum sativum L., Apiales: Apiaceae), marigold (Tagetes erecta L., Asterales: Asteraceae), sweet alyssum (Lobularia maritima L., Brassicales: Brassicaceae), and alfalfa (Medicago sativa L., Fabales: Fabaceae). Our goal was to better understand this predator's ovipositional behavior in response to different plants and its olfactory response to the aphid-induced volatiles from these plants., Results: We found that this lady beetle did not have any significant oviposition preference among the four plant species, but H. axyridis preferred to lay eggs on the lower surface of leaves, regardless of the plant species. H. axyridis females had a significant preference for aphid-infested marigolds, but were not attracted by any of the other three cover plants or marigolds without aphid damage. Compared to the uninfested marigold plants, the emission of 12 compounds significantly increased on the aphid-infested marigolds, and two of them were attractive to H. axyridis under suitable concentrations., Conclusion: H. axyridis did not show any significant oviposition preference among the four cover crops. Aphid-infested marigolds can attract H. axyridis. Indole and terpinen-4-ol mediated lady beetle attraction. These synomones have potential for manipulating populations of H. axyridis as a component of conservation biological control. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Grasshoppers (Orthoptera: Acridoidea) are one of the most dangerous agricultural pests. Environmentally benign microbial pesticides are increasingly desirable for controlling grasshopper outbreaks in fragile ecosystems. However, little is known about natural pathogens infecting this pest. Here we profile the rich viral communities in forty-five grasshopper species and report 302 viruses, including 231 novel species. Most of the identified viruses are related to other insect viruses, and small RNA sequencing indicates that some are targeted by host antiviral RNA interference (RNAi) pathway. Our analysis of relationships between host phylogeny and virus diversity suggests that the composition of viromes is closely allied with host evolution. Overall, this study is a first extensive exploration of viruses in grasshoppers and provides a valuable comparative dataset of both academic and applied interest., (© The Author(s) 2022. Published by Oxford University Press.)

The gut microbiota of insects plays a vital role in digestion, nutrient acquisition, metabolism of dietary toxins, pathogen immunity and maintenance of gut homeostasis. Bacillus thuringinensis (Bt) poisons target insects through its toxins that are activated in the insect gut. The effects of Bt toxins on gut microbiota of insects and their underlying mechanisms are not well understood. In this study, we found that Cry1Ab/2Ab toxins significantly changed the gut bacterial community's structure and reduced the total load of gut bacteria in the Locusta migratoria. In addition, Cry toxins significantly increased the level of reactive oxygen species (ROS) in the gut of locusts. Our results also showed that Cry1Ab/2Ab toxins induced the host gut's immune response by up-regulating of key genes in the Immune deficiency (IMD) and Toll pathway. RNA interference showed that knocking down Relish could narrow the difference in the load, diversity, and composition in gut bacteria caused by Cry toxins. Our findings suggest that Bt potentially influences the gut bacterial community of L. migratoria through host immune response., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The orange wheat blossom midge ( Sitodiplosis mosellana Géhin) is an insect pest that feeds on wheat ( Triticum aestivum L.). The resistance mechanisms of wheat to S. mosellana infestation are largely unknown. In this study, the wheat varieties LX99 and 6218 were identified as highly resistant and susceptible, respectively, via field investigations conducted over two consecutive years. Morphological and microstructural observations of mature wheat kernels following S. mosellana infestation revealed that the degree of cell structure damage in resistant LX99 grains was less than that in susceptible 6218 grains. Transcriptomic and metabolomic analyses of seeds following S. mosellana feeding showed that the differentially expressed genes and differentially accumulated metabolites from LX99 were mostly enriched in several primary and secondary metabolic pathways, including phenylpropanoid biosynthesis, flavonoid biosynthesis, and phenylalanine biosynthesis. Additionally, phenylpropanoid- and flavonoid-related gene expression was significantly upregulated following S. mosellana infestation in LX99 relative to that in 6218. Some metabolites involved in phenylpropanoid/flavonoid pathways, such as cinnamic acid, coumarin, epigallocatechin, and naringenin, were only induced in infested LX99 kernels. These results suggest that phenylpropanoid/flavonoid pathways play important roles in wheat kernel resistance to S. mosellana attack and provide useful insights for the breeding and utilization of resistant varieties.

Odorant receptors (ORs) of ladybird Hippodamia variegata play vital chemosensory roles in searching and locating preys. In the current study, 37 ORs were initially identified from the antennal transcriptome of H. variegata . The quantitative polymerase chain reaction demonstrated that several HvarORs including HvarOR25 were specific or enriched in ladybird antennae. In two-electrode voltage clamp recordings, recombinant HvarOR25 was narrowly tuned to six chemical ligands including aphid-induced, aphid-derived, and plant-derived volatiles. In electroantennogram assays, all six volatiles elicited electrophysiological responses. Among the six volatiles, cis -3-hexenyl acetate, hexyl butyrate, hexyl hexanoate, and 3-methyl-3-buten-1-ol were attractive for both sexes of H. variegata . Additionally, molecular docking indicated that HvarOR25 was bound to all ligands with high binding affinities. Taken together, HvarOR25 facilitates perception of preys by recognizing relevant allelochemicals from hosts and habitat. Our findings provide valuable insights into understanding biological functions of HvarORs and help to develop a novel biocontrol strategy based on olfactory-active compounds.

Natural enemies play an important role in managing insect pests. Orius sauteri (Poppius) (Hemiptera: Anthocoridae), a predator of many soft-body insects, is an important biological control agent in Asia. Understanding this predator's egg-laying preferences and a habitat needs is important for its success in pest control. We investigated the plant acceptability and ovipositional preference of O. sauteri for coriander ( Coriadrum sativum L., Apiales: Apiaceae), marigold ( Tagetes erecta L., Asterales: Asteraceae), sweet alyssum ( Lobularia maritima L., Brassicales: Brassicaceae), and alfalfa ( Medicago sativa L., Fabales: Fabaceae), and focused on the effects of plant physical and anatomical characteristics on the ovipositional preference of O. sauteri . The results showed that O. sauteri can lay eggs on uninfested plants in the vegetative stage and their eggs hatched normally. Orius sauteri females prefer plants with high stomatal density, a large stomatal area, and fewer trichomes as oviposition hosts, and the depth of egg placement was determined by leaf thickness. Our studies suggested that O. sauteri females can select oviposition hosts and specific oviposition sites by assessing the structural qualities of plant surface. Coriander and marigold are potentially suitable host plants for O.sauteri. The results aid the selection of cover crops to enhance natural enemies in the fields.

Microbial communities in insects are related to their geographical sources and contribute to adaptation to the local habitat. The Colorado potato beetle ( Leptinotarsa decemlineata ) (CPB) is a potato pest that causes serious economic losses in Xinjiang Uygur Autonomous Region (XJ) and Heilongjiang Province (HL), China. The influence of microorganisms in the invasion and dispersal of CPB is unclear. We studied microbial communities of CPB collected from nine geographic sources in China using high throughput sequencing technology. Bacteroidetes, Firmicutes, and Proteobacteria were the most dominant phyla, Clostridia, Bacteroidetes, and γ-Proteobacteria were the most dominant classes, Enterobacterales, Lactobacillales, Clostridiales, and Bacteroidales were the most dominant orders, and Enterobacteriaceae, Streptococcidae, Verrucomicrobiaceae, and Rikenellaceae were the most dominant families. There were significant differences, among sources, in the relative abundance of taxa at the genus level. A total of 383 genera were identified, and the dominant bacteria at the genus level were compared between XJ and HL. Pseudomonas was the unique dominant microorganism in the HL area, and the other four microorganisms ( Lelliottia , Enterococcus , Enterobacter , and Lactococcus ) were common within the 2 regions. Bacterial community diversity in CPB from Urumqi, Jimunai, and Wenquan was higher than diversity in other regions. T-Distributed Stochastic Neighbor Embedding (tSNE) analysis indicated that order and genus were appropriate taxonomic levels to distinguish geographical sources of CPB. These findings provide insight into the diversity of microorganisms of CPB in the differences among geographically isolated populations., Competing Interests: YW was employed by company Biowavelet Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Yu, Wang, Li, Yu, Shi and Zhai.)

Background: Thrips, Megalurothrips usitatus, usually display aggregation behavior, which is probably mediated by a male-produced aggregation pheromone. Aggregation pheromones are species-specific, and can be used to develop commercial lures for monitoring and mass-trapping of pests. The active components of the aggregation pheromone for four thrips species have been identified. However, the components of M. usitatus-produced aggregation pheromone are still not clear., Result: Y-tube olfactometer assays showed that both male and virgin female M. usitatus were significantly attracted to male but not female volatiles. This was additionally supported by electroantennogram (EAG) assays. Coupled gas chromatography-electroantennogram detection (GC-EAD) showed that one component of male-specific odors elicited a significant electrophysiological response. This compound was characterized as (2E,6E)-farnesyl acetate, which is structurally different from the active components of the aggregation pheromones of other reported thrips species. Electroantennal responses of M. usitatus increased with increasing doses of synthetic (2E,6E)-farnesyl acetate. Additionally, this compound significantly attracted adults in laboratory behavioral bioassays. Under field conditions, sticky traps with synthetic (2E,6E)-farnesyl acetate caught 1.5-7-fold more M. usitatus than controls, and this effect of the compound at a dose of 60 μg lasted at least 6 days., Conclusion: (2E,6E)-Farnesyl acetate was identified as the male-produced aggregation pheromone of M. usitatus. It could attract this thrips species under laboratory and field conditions, suggesting considerable potential as a commercial application to control M. usitatus populations. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Transgenic maize expressing the Cry1Ab and Cry2Ab protein simultaneously from Bacillus thuringiensis (Bt-maize) has been grown for farm-scale study to investigate its potential impact to non-target arthropod (NTA). The trials were conducted between Bt maize 2A-7 and its parental line (B73-329) in Beijing, China, over 3 years. Richness (C), Shannon index (H), Pielou index (J), Simpson index (D), and Bray-Curtis index were used to evaluate the population dynamics and biodiversity of the dominant arthropods from per 50 plants in crop field. The mainly abundant groups were Aphidoidea, Araneae, Coccinellidae, Anthocoridae, and Thripidae which represented about 90% of the total number of NTA. Although the abundance of NTA varied from year to year, there is no significant difference between Bt maize and non-Bt maize field. Fluctuations were found at individual sample dates, but the trend of these descriptors remained consistent. Further analysis showed the biodiversity indexes of the dominant arthropods C, H, J, D, and Bray-Curtis dissimilarity between Bt maize producing Cry1Ab and Cry2Ab toxin simultaneously and its parental line had no significant difference except for some sampling dates. These results suggested that Bt maize is compatible with the NTAs and provides further evidence of the ecological impact of genetically modified maize.

Background: Honeydew is valuable food source for predators that can build predator numbers and strengthen biological control. Honeydew excreted by hemipterans often supplements the diets of their predators and parasitoids. However, dense sticky honeydew also creates a difficult foraging environment, potentially limiting predator efficiency., Results: We examined the benefits and costs of honeydew excreted by the pear psylla (Cacopsylla chinensis [Yang and Li]) for its key predator in much of Asia, the anthocorid bug Orius sauteri (Poppius). We found these predators spent more time foraging and laid more eggs in the presence of psyllid honeydew compared to the control. However, predators more often foraged among psylla without honeydew than those coated in honeydew. This suggests that while O. sauteri recognized honeydew as a useful cue to prey presence, the predators were more likely to attack pear psylla lacking the sugary excretion. In foraging trials, honeydew consistently reduced the number of psyllids killed by the predator, suggesting it limited O. sauteri mobility or reduced the nutritional value of psyllids as prey. We also found slowed development, reduced longevity, and reduced fecundity of O. sauteri reared on moth eggs (Sitotroga cerealella [Olivier]) coated in honeydew compared to those reared on moth eggs alone., Conclusion: Altogether, our results suggest that psyllid honeydew could serve as a prey-location and oviposition cue for O. sauteri. However, honeydew also limited predator foraging with the potential to limit biological control. More generally, honeydew might form an important type of defense for stationary feeders like psyllids. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Background: Neuropeptide F (NPF) is an intercellular signaling molecule that mediates many physiological and behavioral processes. However, the function of neuropeptide F in mediating the feeding behavior of Locusta migratoria has been unclear., Results: The neuropeptide F 1 precursor cDNA from L. migratoria was obtained and analyzed, and its amino acid sequence deduced. Mature LmiNPF1 was composed of 36 amino acids and was similar to that of Schistocerca gregaria. The spatial and temporal expression profiles of LmiNPF1 were investigated. LmiNPF1 was primarily expressed in the central nervous system, especially in the brain, and the expression levels were higher during the day than during the night. However, starvation activated LmiNPF1 expression increases, and downregulation of LmiNPF1 inhibited locust feeding behavior., Conclusion: LmiNPF1 promotes the feeding behavior of the locust and is a potential molecular target to control locust feeding. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Pear psylla, Cacopsylla chinensis (Yang & Li) are present as two seasonal morphotypes in pear orchards where they, suck phloem sap, defoliate pear trees, and cause fruit russet. Despite the importance of natural enemies in psyllid control, the interactions between predators and the two seasonal morphotypes of psyllids remain poorly documented. Here we determined the predation efficiencies of the Asian lady beetle, Harmonia axyridis (Pallas) on the two psyllid morphotypes. Predation of H. axyridis on both morphotypes conformed to a Type II functional response: the proportion of consumed psyllids decreased with increasing prey densities. Predation efficiency of H. axyridis against the winterforms increased with temperature when measured from 8 to 25°C. Predation rate on the summerforms was significantly higher than that of the winterforms. This was linked to smaller body size, higher soluble protein level, thinner cuticle, and lower chitin content of summerform psyllids compared to winterform psyllids. Predation capacities of H. axyridis on both morphotypes indicated its potential as a biocontrol agent in psyllids management. Predation efficiency was higher on summerforms, likely due to the difference in body size, exoskeleton fragility, and nutritional value between the two morphotypes. Due to the Type II functional response of H. axyridis to both morphotypes of pear psylla, application of H. axyridis in pear orchards under suitable temperatures could be taken into consideration for suppression of C. chinensis, although further experiments conducted in field conditions are needed to validate our findings., Competing Interests: The authors have declared that no competing interests exist.

From 2014 to 2016, individuals of the principal non-target arthropod (NTA) species in a field of Bt maize expressing the Cry1Ac protein (Bt38) were compared to those in a control field of the corresponding non-transformed near isoline (Z58). For all 3 years, the population dynamics and biodiversity of NTAs were analyzed to determine if any differences might be attributable to the genetically modified (GM) maize being evaluated. The main NTAs in these fields were Aphidoidea, Pentatomidae,Araneae, and Coccinellidae. Temporal variation in NTA diversity across sample dates within a year showed no significant differences between the Bt maize and the non-Bt maize field in the total number of individuals of the dominant arthropod species per 25 plants, the Shannon index (H), Pielou index (J), Simpson index (D), and Bray-Curtis index. The cultivation of Bt corn failed to show any detrimental evidence on individuals, H, J, D, and Bray-Curtis index of NTAs, and these parameters were identical in Bt and non-Bt corn plots. These results provide further evidence of the lack of ecological impact of GM maize.

Aphids are important agricultural pests because of their feeding and, for many species, their ability to transmit plant pathogenic viruses. Orius sauteri (Poppius) (Hemiptera: Anthocoridae) is an important predator in northern China of many aphid species, including Aphis craccivora (Koch) (Hemiptera: Aphididae). We measured the functional response and intraspecific competition of O. sauteri under different densities of A. craccivora at several constant temperatures. Predation of O. sauteri on A. craccivora conformed to Holling's type II functional response at all temperatures tested. The predation capability of O. sauteri showed a significant correlation with temperature. The predation capacity of O. sauteri was highest at 25°C, at which temperature we found the highest instantaneous attack rate (a = 0.8711), the theoretical maximum number of aphids captured (181.36), and the shortest handling time (Th = 0.0055). Predation by O. sauteri increased progressively with temperature from 15 to 25°C, but then decreased at 30°C. Intraspecific competition of O. sauteri increased positively with predator density under all temperatures examined. Our results indicate O. sauteri has significant potential for use as a biocontrol agent, under a wide range of temperatures. The inverse density-dependent predation and intraspecific competition of O. sauteri suggests that to obtain the greatest degree of pest suppression, releases of O. sauteri should begin early in the cropping season and numbers released should take prey density into consideration.

Isaria farinosa is a pathogen of alpine Thitarodes larvae that are hosts for the Chinese medicinal fungus, Ophiocordyceps sinensis. A matrix analysis indicated that the optimal culture conditions for the mycelial growth of I. farinosa are a 50-mL liquid broth in a 250-mL flask at more than 100-rpm rotation and 15-25 °C. Illumination does not affect the mycelial growth. The optimal nutrition requirements are D-(+)-galactose and D-(-)-fructose as carbon resources and D-cysteine as well as yeast powder, peptone, and beef extract as nitrogen resources at a carbon-to-nitrogen ratio of 1:1 to 1:7. The mineral component and vitamins also significantly increase the mycelial growth of I. farinosa. Based on the optimal culture conditions and nutrition requirements for the mycelial growth of I. farinosa, the effects of altitude on mycelial growth and its metabolome were evaluated using quadrupole-time-of-flight/mass spectrometry, principal component analysis and partial least squares discriminant analysis. The altitude did not affect the mycelial production but significantly regulated its metabolome. The study presents a new approach to better select a method for producing more useful metabolites and highlights the necessity of establishing standards for culturing methods related to altitude to preserve fungal quality; additionally, the results indicate that the use of a fermenter may meet the demands of large-scale mycelial production.

Paranosema locustae is a gut parasite that has been applied widely in the control of grasshoppers in many parts of the world. Usually, P. locustae is transmitted horizontally via passive modes under natural conditions but in the current study, a positive transmission strategy of P. locustae was demonstrated. First, infection by P. locustae resulted in the cuticula of infected Locusta migratoria nymphs to become lighter in color: normally only a small proportion of locusts are pale with most either being partly or mostly black; but locusts infected with P. locustae became pale. And it was found that the change to pale occurred even among uninfected black and partly black nymphs reared with infected locusts. The eumelanin of the thorax and abdomen of infected individuals decreased significantly, as did the level of dopamine. In addition, there was a decrease in phenol oxidase activity and the expression of henna and pale , which are involved in the synthesis of cuticle melanin, decreased. What is the ecological significance of this increase in light-colored hosts caused by P. locustae ? We discovered that light-colored locusts were more susceptible to the microsporidian pathogen than dark-colored individuals were, because of their weaker melanization. Phenol oxidase activity in pale locusts was lower than that of black locusts, but the serpin expression level of pale locusts was higher than that of black individuals. When examined for infection, it was found that initially uninfected nymphs had picked up P. locustae infections indicating that infections are readily passed from one pale locust to another. The infection rate of healthy locusts reared with light-colored locusts infected with P. locustae was 100% which was more than with black-colored ones. The increase in albinistic locusts clearly promoted the prevalence of P. locustae in the total population. In conclusion, these results elucidated a new strategy of positive self-transmission in P. locustae . Importance : Mother Nature always grants wisdom to her creatures and feeds them carefully. This wisdom is particularly apparent in the relationships between two interacting species. In this study, our team focused on the interaction between L. migratoria and P. locustae . In a previous study, it was found that L. migratoria isolate infected individuals, reducing avoiding the spread of P. locustae , in a previous study. The solitary, pale individuals infected by P. locustae were left behind as locust groups marched ahead, leading to a kind of behavioral immunity in the insects. Here, we reported that P. locustae promotes pigmentation loss in L. migratoria , causing a larger proportion of light-colored individuals, and these lighter individuals which possessed weaker immunity against pathogens. This strategy is advantageous to P. locustae , as it promotes its propagation and spread. These extraordinary abilities of L. migratoria and P. locustae have accumulated over millennia of years of interaction.

Locusta has strong fly wings to ensure its long distance migration, but the molecular mechanism that regulates the Locusta wing development is poorly understood. To address the developmental mechanism of the Locusta flying wing, we cloned the Dpp target gene spalt (sal) and analyzed its function in wing growth in the Locusta. The Locusta wing size is apparently reduced with vein defects when sal is interfered by injection of dsRNA, indicating that sal is required for locust wing growth and vein formation. This function is conserved during the Drosophila wing development. To better understand sal's function in wing growth, we then used Drosophila wing disc as a model for further study. We found that sal promotes cell proliferation in the whole wing disc via positive regulation of a microRNA bantam. Our results firstly unravel sal's function in the Locusta wing growth and confirm a highly conserved function of sal in Locusta and Drosophila.

Based on MS and NMR data and bioassay-guided tracing, three insecticidal alkaloids I, II and III from Cynanchum mongolicum were identified to be antofine N-oxide, antofine and tylophorine. Alkaloid I was more toxic than alkaloids II and III, but they were less active against Spodoptera litura than total alkaloids. The contact toxicity from these alkaloids against the aphid Lipaphis erysimi was significant, as the 24 h-LC50 values of alkaloids I, II, III and total alkaloids were 292.48, 367.21, 487.791 and 163.52 mg/L, respectively. The development disruption of S. litura larvae was tested, the pupation and emergence rates of S. litura decreased and the acute mortality of S. litura increased significantly by day 3 after being injected in their body cavity with 10-40 mg/L of total alkaloid. The ecdysone titer of treated S. litura larvae and prepupae declined with increasing alkaloid concentration. The alkaloids of Cynanchum mongolicum are potential insect growth inhibitors.

Transgenic corn producing Cry1Ac toxins from Bacillus thuringiensis (Bt) provides effective control of Asian corn borer, Ostrinia furnacalis (Guenée), and thus reduces insecticide applications. However, whether Bt corn exerts undesirable effects on non-target arthropods (NTAs) is still controversial. We conducted a 2-yr study in Shangzhuang Agricultural Experiment Station to assess the potential impact of Bt corn on field population density, biodiversity, community composition and structure of NTAs. On each sampling date, the total abundance, Shannon's diversity index, Pielou's evenness index and Simpson's diversity index were not significantly affected by Bt corn as compared to non-Bt corn. The "sampling dates" had a significant effect on these indices, but no clear tendencies related to "Bt corn" or "sampling dates X corn variety" interaction were recorded. Principal response curve analysis of variance indicated that Bt corn did not alter the distribution of NTAs communities. Bray-Curtis dissimilarity and distance analysis showed that Cry1Ac toxin exposure did not increase community dissimilarities between Bt and non-Bt corn plots and that the evolution of non-target arthropod community was similar on the two corn varieties. The cultivation of Bt corn failed to show any detrimental evidence on the density of non-target herbivores, predators and parasitoids. The composition of herbivores, predators and parasitoids was identical in Bt and non-Bt corn plots. Taken together, results from the present work support that Bt corn producing Cry1Ac toxins does not adversely affect NTAs.

Locusts are infamous for their ability to aggregate into gregarious migratory swarms that pose a major threat to food security. Aggregation is elicited by an interplay of visual, tactile, and chemical stimuli, but the aggregation pheromone in feces is particularly important. Infection by the microsporidian parasite Paranosema (Nosema) locustae is known to inhibit aggregation of solitary Locusta migratoria manilensis and to induce gregarious locusts to shift back to solitary behavior. Here we suggest that P. locustae achieves this effect by acidifying the hindgut and modulating the locust immune response, which suppresses the growth of the hindgut bacteria that produce aggregation pheromones. This in turn reduces production of the neurotransmitter serotonin that initiates gregarious behavior. Healthy L. migratoria manilensis exposed to olfactory stimuli from parasite-infected locusts also produced significantly less serotonin, reducing gregarization. P. locustae also suppresses biosynthesis of the neurotransmitter dopamine that maintains gregarization. Our findings reveal the mechanisms by which P. locustae reduces production of aggregation pheromone and blocks the initiation and maintainence of gregarious behavior., Competing Interests: The authors declare no conflict of interest.

The relatively low direct mortality caused by Paranosema locustae (Canning) has limited its application for controlling grasshopper when densities are high, and this study sought to determine if the simultaneous use of this pathogen and the IGR, Flufenoxuron (Cascade) could provide effective control. Nine treatments were tested: 45% Malathion EC at 1500 ml/ha, 5% Cascade at 150 ml/ha, 5% Cascade at 75 ml/ha, 5% Cascade at 37.5 ml/ha, P. locustae at 7.5 x 10(9) spores/ha, combinations of 5% Cascade at 75 ml/ha and P. locustae at 7.5 x 10(9) spores/ha, applied in different rations (1:1, 1:2, 1:3) in the same plot, the untreated control. P. locustae was applied on nonoverlapping plots with the IGR. The different in-plot combinations of P. locustae and Cascade in different ratios provided significantly better overall control of grasshoppers (all species) than the treatment of 5% Cascade of 150 ml/ha after 5d, but combinations were not significantly different from the other concentrations of Cascade after 12 and 31 d. When results were examined separately for specific species of grasshoppers, reduction of Dasyhippus harbipes (Fischer-Waldheim), was higher than that of Myrmeleotettix palpalis (Zubovsky). While combinations showed significant differences in the infection of different grasshopper species at 5 and 12 d posttreatment, no significant differences in rate of infection among the primary species (M. palpalis, D. harbipes, and Oedaleus asiaticus Bei-Bienko) were detected 31 d posttreatment. Our study found that P. locustae by itself could control grasshopper populations at medium densities but the combined application of P. locustae and Cascade at a ratio of 1:2 was more effective against high-density grasshopper populations.

The acute lethal toxicity of the extracted and purified gross alkaloids from Cynanchum komarovii has been demonstrated on the insect pest, Spodoptera litura. The toxic regression equation of the gross alkaloids for S. litura larvae was Y = -2.69701 + 0.78711X and the LC(50) value 2669.88 mg L(-1). The growth disruptions of S. litura caused by the alkaloids from C. komarovii were also evaluated. The rates of growth inhibition, malformation, developmental duration (from third instar to pupation), pupation and emergence inhibition of S. litura caused by the gross alkaloids at dose rates between 50 and 800 mg L(-1) were 30.13-91.71%, 0.00-20.00%, 12-72 h, 16.66-36.66% and 16.66-50.00%, respectively. The alkaloids caused the modification of the cuticular components of S. litura. The relative contents of cuticular lipids, proteins and chitins showed a 3.02-17.11% rise before exposure, followed by 2.29-12.96% and 0.75-4.13% declines 72 h after treatment with gross alkaloids with concentrations between 50 and 800 mg L(-1). Initial studies on the insecticidal properties of C. komarovii gross alkaloids indicate these to be very potent insect growth inhibitors.

Rapidly emerging insecticide resistance is creating an urgent need for new active ingredients to control the adult mosquitoes that vector malaria. Biopesticides based on the spores of entomopathogenic fungi have shown considerable promise by causing very substantial mortality within 7-14 days of exposure. This mortality will generate excellent malaria control if there is a high likelihood that mosquitoes contact fungi early in their adult lives. However, where contact rates are lower, as might result from poor pesticide coverage, some mosquitoes will contact fungi one or more feeding cycles after they acquire malaria, and so risk transmitting malaria before the fungus kills them. Critics have argued that 'slow acting' fungal biopesticides are, therefore, incapable of delivering malaria control in real-world contexts. Here, utilizing standard WHO laboratory protocols, we demonstrate effective action of a biopesticide much faster than previously reported. Specifically, we show that transient exposure to clay tiles sprayed with a candidate biopesticide comprising spores of a natural isolate of Beauveria bassiana, could reduce malaria transmission potential to zero within a feeding cycle. The effect resulted from a combination of high mortality and rapid fungal-induced reduction in feeding and flight capacity. Additionally, multiple insecticide-resistant lines from three key African malaria vector species were completely susceptible to fungus. Thus, fungal biopesticides can block transmission on a par with chemical insecticides, and can achieve this where chemical insecticides have little impact. These results support broadening the current vector control paradigm beyond fast-acting chemical toxins.

Based on the data obtained from laboratory and field experiments, this paper established the morphological and behavioral indices for phase solitaria (S) and phase gregaria (G) of oriental migratory locust Locusta migratoria manilensis. S-females and S-males jumped 1.4 times x min(-1), and the frequency of turning was 1.3 times x min(-1) for S-females and 1.4 times x min(-1) for S-males. G-females and G-males jumped 1.6 times x min(-1), and the frequency of turning was 1.6 times x min(-1) for G-females and 1.5 times x min(-1) for G-males. Both jumping and turning behaviours were significantly greater (P<0.05) for gregaria locusts than for solitairia locusts. These behavioral parameters of the frequency of locust jumping and turning can be used as "the behavioral indices for phase transformation". The F/C ratio and the E/F ratio might be used as the morphological indices for phase transformation of the gregaria and solitaria of Locusta migratoria manilensis. The F/C ratio goes up as the locust grows, but the F/C ratio of females from the 4th instar to adult is less than that of the male of the same instar and phase. Comparing with the F/C and E/F ratio of the male, those of the female exhibited significant difference in the same phase. The F/C ratio of the fourth instar, fifth instar and adults were 2.5, 2.8 and 3.3 for S-females, and 2.6, 2.9 and 3.5 for S-males. As for G-females and and G-males, the F/C ratio of the fourth instar, fifth instar and adults were 2.5, 2.8, 3.3 and 2.5, 2.7, 3.1, respectively. The E/F ratios of adults can be used as "the morphological phase indices for phase transformation".